1 NHNYPPLSLLTF(1)[2.871] SHDWRLMLSHLQ(1)[2.779] QHDRLLMLSYLV(1)[2.666] HHILPPKALLFG(1)[2.623] SHTFPPSWLLVR(1)[2.509] YSVKPPLPYLPP(1)[2.536] H-x(2)-P(2)-x(2)-L(2) 6 Phage display (common panning) 3/4 PMID:17881351 Anti-Ogawa O-antigen monoclonal antibody S-20-4 O-specific polysaccharide of LPS Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance was measured at 450 nm with a kinetic microplate reader (Molecular Devices). Phage were incubated with the selection Ab in solution, followed by affinity capture of the Ab-phage complexes onto protein A/G-agarose. In panning, phage bound to resin were eluted with of glycine HCl (pH 2.2) and rapidly neutralized with Tris-HCl (pH 9.1). Each of the six phage clones that could bind to both monoclonal antibodies also competed with LPS for binding to S-20-4, suggesting that the peptides bind close to the paratope of the Ab. In order to predict how these peptide mimics interact with S-20-4 compared with its carbohydrate counterpart, one peptide mimic, 4P-8 (NHNYPPLSLLTF), which is one of the highest affinity binders and shares motifs with several other peptide mimics, was selected for further studies using computer modeling methods and site-directed mutagenesis. These studies suggest that 4P-8 is recognized as a hairpin structure that mimics some O-SP interactions with S-20-4 and also makes unique ligand interactions with S-20-4. In addition, 4P-8-KLH was able to elicit anti-LPS Abs in mice, but the immune response was not vibriocidal or protective. However, boosting with 4P-8-KLH after immunizing with LPS prolonged the LPS-reactive IgG and IgM Ab responses as well as vibriocidal titers and provided a much greater degree of protection than priming with LPS alone. 2 NHNYPPLSLLTF(1)[2.871] SHKIHPFQTTPY(1)[0.851] HSPRYHSEPQFG(1)[2.118] HAPRYHTIYGWP(1)[1.976] H-x(2)-P(2)-x(2)-L(2), S-H-[KR]-[LI], H-x-P-R-x-H 4 Phage display (common panning) 3/4 PMID:17881351 Anti-Ogawa O-antigen monoclonal antibody S-20-4 O-specific polysaccharide of LPS Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance was measured at 450 nm with a kinetic microplate reader (Molecular Devices). Phage were incubated with the selection Ab in solution, followed by affinity capture of the Ab-phage complexes onto protein A/G-agarose. In panning, phage bound to resin were (nonelution) directly added to a culture of E. coli strain ER2738 for amplification. Amplified phage were purified by precipitating the phage with polyethylene glycol/NaCl. In order to predict how these peptide mimics interact with S-20-4 compared with its carbohydrate counterpart, one peptide mimic, 4P-8(NHNYPPLSLLTF), which is one of the highest affinity binders and shares motifs with several other peptide mimics,was selected for further studies using computer modeling methods and site-directed mutagenesis. These studies suggest that 4P-8 is recognized as a hairpin structure that mimics some O-SP interactions with S-20-4 and also makes unique ligand interactions with S-20-4. In addition, 4P-8-KLH was able to elicit anti-LPS Abs in mice, but the immune response was not vibriocidal or protective. However, boosting with 4P-8-KLH after immunizing with LPS prolonged the LPS-reactive IgG and IgM Ab responses as well as vibriocidal titers and provided a much greater degree of protection than priming with LPS alone. 3 SHKLHKV(1)[0.526] SHRLPLK(1)[1.350] SHRLPAK(1)[1.075] SHRLPVK(1)[1.115] S-H-[KR]-[LI] 4 Phage display (common panning) 3/4 PMID:17881351 Anti-Ogawa O-antigen monoclonal antibody S-20-4 O-specific polysaccharide of LPS Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Absorbance was measured at 450 nm with a kinetic microplate reader (Molecular Devices). Phage were incubated with the selection Ab in solution, followed by affinity capture of the Ab-phage complexes onto protein A/G-agarose. In panning, phage bound to resin were eluted with of glycine HCl (pH 2.2) and rapidly neutralized with Tris-HCl (pH 9.1). 4 HAPRWHL(2)[1.935/2.274] HAPRWHW(1)[1.421] HAPRYHY(1)[1.656] HSPRWHL(1)[2.177] HSPRYHM(1)[1.288] H-x-P-R-x-H 5 Phage display (common panning) 3/4 PMID:17881351 Anti-Ogawa O-antigen monoclonal antibody S-20-4 O-specific polysaccharide of LPS Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Absorbance was measured at 450 nm with a kinetic microplate reader (Molecular Devices). Phage were incubated with the selection Ab in solution, followed by affinity capture of the Ab-phage complexes onto protein A/G-agarose. In panning, phage bound to resin were (nonelution) directly added to a culture of E. coli strain ER2738 for amplification. Amplified phage were purified by precipitating the phage with polyethylene glycol/NaCl. 5 NPEDCFKTGCNSPT(6) LMEPCYAWGCTGPK(3) IMEQCFEIGCPATN(2) NDSQCWKLGCIAEK(1) SNVECDGLNCDWII(1) N-x-E-x(2)-[FYW]-x(2)-G 5 Phage display (subtractive panning) 3 PMID:15034020 Anti-IA-2 monoclonal antibody 96/3 Receptor-type tyrosine-protein phosphatase-like N (R-PTP-N) Not determined. Not determined. f88-4/15mer and f88-4/Cys4 phage display library pool The two libraries were mixed and phage reactive to bovine IgG was removed by incubation in tubes coated with purified bovine IgG. The predicted surface on a three-dimensional homology model of the tyrosine phosphatase domain of IA-2 was analyzed for clusters of Asn, Glu, and aromatic residues and amino acids contributing to the epitope investigated using site-directed mutagenesis. Mutation of each of amino acids Asn(858), Glu(836), and Trp(799) reduced 96/3 Ab binding by >45%. Mutations of these residues also inhibited binding of serum autoantibodies from IA-2 Ab-positive type 1 diabetic patients. This study identifies a region commonly recognized by autoantibodies in type 1 diabetes that overlaps with dominant T cell determinants. 6 EIDAPRVFDARFSRHYSVQL[0.47 ± 0.07] LAVFRPWKARNNGTNEYRSR[0.81 ± 0.07] GAEASTSLFHFRIPRSYFLR[0.73 ± 0.02] FCSLTSLATSCGAAVFIPFR[0.72 ± 0.02] 4 Phage display (subtractive panning) 4 PMID:14742560 Anti-AMA1 monoclonal antibody 4G2dc1 Apical membrane antigen 1, AMA1 Not determined. Not determined. X20 fUSE5 phage display library ELISA The absorbance value was measured at 490 nm. All ELISAs were performed in duplicate. Data shown were reproduced from the graph and expressed as the means ± individual values. 4G2dc1-coated wells were blocked with 5% BLOTTO (milk powder diluted in phosphate-buffered saline [PBS]), and the 20-mer phage peptide library was preincubated in 1% BLOTTO for 15 min to remove any milk-binding phage before being added to the 4G2dc1-coated wells. 7 WGILSRAATNSGWVGLIPHK[0.81 ± 0.28] 1 Phage display (subtractive panning) 5 PMID:14742560 Anti-AMA1 monoclonal antibody 4G2dc1 Apical membrane antigen 1, AMA1 Not determined. Not determined. X20 fUSE5 phage display library ELISA The absorbance value was measured at 490 nm. All ELISAs were performed in duplicate. Data shown were reproduced from the graph and expressed as the means ± individual values. 4G2dc1-coated wells were blocked with 5% BLOTTO (milk powder diluted in phosphate-buffered saline [PBS]), and the 20-mer phage peptide library was preincubated in 1% BLOTTO for 15 min to remove any milk-binding phage before being added to the 4G2dc1-coated wells. 8 IPSTAFTDIAWVRLPNHY[1.07 ± 0.02] LASLRKAFADTVPVRPPSNY[0.90 ± 0.01] FGGVSNEDVPSVHFKPPSSR[0.89 ± 0.04] A-F-x-D-x(3)-V-R-x-P-x(2)-Y 3 Phage display (subtractive panning) 6 PMID:14742560 Anti-AMA1 monoclonal antibody 4G2dc1 Apical membrane antigen 1, AMA1 Not determined. Not determined. X20 fUSE5 phage display library ELISA The absorbance value was measured at 490 nm. All ELISAs were performed in duplicate. Data shown were reproduced from the graph and expressed as the means ± individual values. 4G2dc1-coated wells were blocked with 5% BLOTTO (milk powder diluted in phosphate-buffered saline [PBS]), and the 20-mer phage peptide library was preincubated in 1% BLOTTO for 15 min to remove any milk-binding phage before being added to the 4G2dc1-coated wells. The three peptides elicited antibody responses in rabbits that recognized the peptide immunogen and both recombinant and parasite AMA1. Human antibodies in plasma samples from individuals exposed to chronic malaria reacted with J1 (IPSTAFTDIAWVRLPNHY) and J7 (LASLRKAFADTVPVRPPSNY) peptides and were isolated using immobilized peptide immunoadsorbents. Both rabbit and human antibodies specific for J1 and J7 peptides were able to inhibit the invasion of erythrocytes by P. falciparum merozoites. 9 AFDWTFVPSLIL(17) SLDWSHVPLLVL(3) GPFINKSLGSSP(2) LDVRPWYVTPLP(2) LEPPQLWWPWEH(2) WSSNLRVLPWSP(1) SLYVAPWWDDPP(1) QLVIFPWNVTVP(1) MLTVEPWTISNT(1) 9 Phage display (subtractive panning) 4 PMID:16960395 C-C chemokine receptor type 5, CCR5 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In this study, CCR5-expressing CHO cells (CHO/CCR5 cells) were used to select CCR5-binding peptides from a phage-displayed 12-mers peptide library. According to the paper, the peptide AFDWTFVPSLIL was specific for CCR5 and that it might become a CCR5 antagonist. 10 CLSLMPRLC[1.795 ± 0.192] CNPMMRPLC[1.922 ± 0.105] CQMRTTIRC[1.662 ± 0.125] CRLMMLQQC[1.629 ± 0.218] CMLLQNRQC[0.997 ± 0.157] CTLQASILC[1.011 ± 0.176] CLSLITRLC[0.585 ± 0.088] 7 Phage display (subtractive panning) 4 PMID:15930303 Interleukin-6 receptor subunit alpha Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Individual sIL-6Rα-binding phage clones were tested for their ability to compete with IL-6 protein for binding to immobilized sIL-6Rα. Phages were added to sIL-6Rα-coated wells in a 96-well plate at a concentration of e12 pfu/mL. After 1 hour of incubation with phage, IL-6 protein was added to the wells, and bound IL-6 was then measured using biotin-conjugated anti-IL-6 mAb, HRP-conjugated streptavidin, and TMB substrate. Optical density was quantified at 405 nm. Data were reproduced from the graph and shown as mean of three independent experiments ± SD. The phage library was added to non-bait-coated dishes (those coated with human IL-6Rβ proteins) for preabsorption. According to paper, peptide LSLITRL substantially inhibits IL-6-induced vascular endothelial growth factor-A expression and angiogenesis in different cancer cell lines. 11 YTPPPLIEAFAT 1 Phage display (common panning) 3 PMID:16740692 EWS-Fli1 protein RNA helicase A, RHA Not determined. Not determined. Ph.D.-12 phage display library (X12) Panning was performed on the surface of a well from a 96-well microtiter plate (Costar, Cambridge, MA). By screening, twenty-eight novel peptides that differentially bind to EWS-FLI1 were identified from phage sequencing. Peptide YTPPPLIEAFAT showed significant homology to human RHA (GenBank accession number A47363) at the amino acid 822 to 831 region. 12 CWWEWTATC(11) CWWEWTIGC(3) CWWEWTGAC(2) CQFTVWEWC(1) CCLGRWCWC(1) CWEWSGNGC(1) CSWTSLPFC(1) CHAWMGLRC(1) CFKRSGSLC(1) CWRRGYGGC(1) CKPWFPELC(1) CALMIANSC(1) CRLTANLPC(1) W(2)-E-W-[ST] 13 Phage display (subtractive panning) 4 PMID:16705086 Anti-CD20 monoclonal antibody rituximab B-lymphocyte antigen CD20 2OSL, Not determined. CX7C fUSE5 phage display library In the third and fourth rounds of selection, the library was precleared on the antibody cetuximab (Merck, Darmstadt, Germany). According to the paper, rituximab binds a discontinuous epitope in CD20, comprised of (170)ANPS(173) and (182)YCYSI(185), with both strings brought in steric proximity by a disulfide bridge between C(167) and C(183). 13 CTRGFEWLC(10) CVFYYNWVC(3) CVYLYEWVC(2) CVYHYRWLC(1) CRDRYWPWC(1) CEWIYHYWC(1) [FY]-[EH]-W-[LV] 6 Phage display (subtractive panning) 4 PMID:17440057 Anti-CD25 monoclonal antibody basiliximab Interleukin-2 receptor subunit alpha 3IU3, Not determined. CX7C fUSE5 phage display library In the third and fourth round of selection, the library was precleared on rituximab. A striking amino acid sequence motif was enriched. This motif is homologous to the peptide ERIYHFV comprising amino acid positions 116 to 122 within the extracellular domain of CD25, suggesting that this is the basiliximab epitope. 14 CEFAQC(2) CDWVFC(1) CDFKAC(1) CDFVGC(1) CEYKEC(1) CEENLC(1) CEFLFC(1) CEFFFC(1) CEKFPC(1) CEHYPC(1) CEFESC(1) CPFIRC(1) CPFTEC(1) CPFDRC(1) CPYRHC(1) CPFMAC(1) CPFKEC(1) CPFHTC(1) CPYKQC(1) CPFKQC(1) CPFAAC(1) CPYHEC(1) CPYRRC(1) CPYRFC(1) C-[DEP]-[FYW]-[DEHKR]-x-C 24 Phage display (common panning) 3 PMID:12538646 Baculoviral IAP repeat-containing protein 4 Not determined. Not determined. Not determined. CX4C and X6 phage display library pool Recombinant full-length human XIAP are used to select peptides from phage display peptide libraries. The DNA inserts of 36 randomly chosen phage clones (18 from the X6 library and 18 from the CX4C library) recovered from the third round of biopanning on XIAP were sequenced. A total of 32 phage clones derived from both libraries displayed the consensus motif C-(D/E/P)-(aromatic)-(charged)-xC. Protein-protein interaction assays revealed that caspase-3 and caspase-7 (but not caspase-8) blocked the binding of the CEFESC phage to XIAP, indicating that this peptide targets a domain within XIAP that is related to the caspase-binding site. In fact, the sequence EFES is homologous to a loop unique to the executioner caspase-3 and caspase-7 that are targeted by XIAP. 15 HLANHQGKWRLH(11) LPLFNANGTWQF(6) VELRNLGGTWRP(1) 3 Phage display (common panning) 3 PMID:16496330 Anti-glycoprotein E2 monoclonal antibody AP33 Envelope glycoprotein E2 Not determined. Not determined. Ph.D.-12 phage display library (X12) Nineteen selected by AP33, were analyzed, and of these 17 were reactive to the selecting antibody. Phage selected by MAb AP33 could be divided into three groups based on the amino acid sequence of the random peptide insert. Multiple alignment of the phage sequences showed absolute conservation of four amino acids, namely L413 (Leucine corresponding to position 413 of the H77 E1E2 sequence), N415, G418, and W420. 16 SRVPQHSYWHSN(3) SKSILNSQGSWH(3) TEPWLPSNPSWA(1) STLPTFPRWMTD(1) YEPGPAWRFHRE(1) RNTHINVDGQYH(1) VHSPNPYWGPTL(1) HKTLNMASYWHN(1) GRPIFNEQAEYL(1) DARPIVGPFLLH(1) SIPTGSSWYALA(1) SPTTFGQAWHTV(1) THPIISTEASWH(1) YTQTGWLNWHER(1) NPHNNNYWGPRI(1) VYPQNKHHTWYS(1) NIWPKQSPYYTS(1) QYRANESSRWHV(1) SFPNQSIHRSTW(1) 19 Phage display (common panning) 3 PMID:16496330 Anti-glycoprotein E2 monoclonal antibody 3/11 Envelope glycoprotein E2 Not determined. Not determined. Ph.D.-12 phage display library (X12) 17 GRYFPDSPAWNH(3) NPHNNNYWGPRI(2) NTGWLPKFPAWT(1) SQVWNTNPEYLT(1) SYTFWPDSYYSS(1) IPNAWNPRAPWQ(1) SKSILNSQGSWH(1) 7 Phage display (common panning) 4 PMID:16496330 Anti-glycoprotein E2 monoclonal antibody 3/11 Envelope glycoprotein E2 Not determined. Not determined. Ph.D.-12 phage display library (X12) 18 WGQQPIDPALWM(2) FQQPLDPWTSPI(2) WQHPLHDWFDLV(2) SNGNWQFPVDPS(1) HVWQGPVDPATF(1) QSWWQQPFDPMD(1) SPWQQWQMPVDV(1) LQTPIDWDEPWF(1) HQAPLDLDWWVD(1) SQLPLDLWMFPD(1) MWQKPLVVHWPT(1) WQMPLTTELDSP(1) NTHQYPIYYPWR(1) HQHPLNNWPNIT(1) SPQVPLTTWNYH(1) WHSQTPVYYWHT(1) TQSPMWWDDFFF(1) IQPQQYWDPFRY(1) FNPQKFWDPYLD(1) NQQAAWDPWMLM(1) HQSYVDPWMLDH(1) WQFHMDPWSLHQ(1) REQLYLDYNVFS(1) QQSRTDPWLPVP(1) WQLHYEPFSVNP(1) NGGQVALSPDWL(1) THMYQSIYVPDI(1) YPYQHHLTYNMW(1) NIPQEHMKFFMA(1) VPQEVWSFFSIP(1) SQQELMLWFNTQ(1) VPTWQQSVPYDL(1) HQIETLLHWFKP(1) GPQQYVNALLFY(1) THQQALPWADMF(1) YQQHINWHSWDT(1) Q-x-P-[AVLIFYW]-D-P(0,1), Q-x-P-[AVLIFYW], Q-x(2)-[AVLIFYW]-D-P 36 Phage display (common panning) 4 PMID:16636049 Protein-glutamine gamma-glutamyltransferase 2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) This study further confirmed that the sequences were favored for transamidation using modified glutathione S-transferase (GST) for recombinant peptide-GST fusion proteins. Most of the fusion proteins exhibited a considerable increase in incorporation of primary amines over that of modified GST alone. 19 EQYQLAWPAMQS(3) DQMMLPWPAVKL(2) SQHPLPWPVLML(2) WQHKIDLRYNGA(2) WHYPPPYDLMRT(2) LDQIVIPWPTPR(1) DQWMMAWPSLTL(1) SEQHLLKWPPWH(1) EQQQLSWPPFLP(1) SQIPMAWPLLSL(1) TQYTMTWPFSWR(1) WQQQMKWPSFTY(1) WQIPVDWPPLPP(1) WQMQLPWPPRLS(1) GQMILPWPSPRA(1) EQFPIAFPRYSI(1) HQMVLTYPWVPT(1) DQYVLTFPWTPR(1) WQMPVNLPIRYP(1) WQLKYPWISPVG(1) HQIPIQIHSFAM(1) DQSKVWLLPSWK(1) WHYQVPQWLYRH(1) WYHPLQLTPAPI(1) WSHMLYLPGSAP(1) WSHPMYLGPSAL(1) YNAPSSFIPLVM(1) NHPIYPHMLLSY(1) WDSHILTYHVPY(1) Q-x(2)-[AVLIFYW]-x-W-P 29 Phage display (common panning) 4 PMID:16636049 Coagulation factor XIII A chain Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) This study further confirmed that the sequences were favored for transamidation using modified glutathione S-transferase (GST) for recombinant peptide-GST fusion proteins. Most of the fusion proteins exhibited a considerable increase in incorporation of primary amines over that of modified GST alone. 20 CSNSDKPKC(2) CYNSDKPKC(1) CRNSDKPKC(1) CSDSDKPKC(1) CWESDKPKC(1) CKWSDKPKC(1) 6 Phage display (common panning) 3 PMID:16988228 Anti-AMA1 monoclonal antibody 45B1 Apical membrane antigen-1 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The MAb 45B1 epitope is conserved and located within the AMA1 domain II loop. 21 CFYEMPTRC(5) CDHFGMHRC(3) CTPTFRANC(1) CTSLWALQC(1) 4 Phage display (common panning) 5 PMID:16988228 Anti-P235 rhoptry protein monoclonal antibody 25.77 P235 rhoptry protein Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The epitope recognized by 25.77 could not be assigned. 22 HMSEYPW(4) CMTCLVK(3) IRPMVYS(3) IRPMVDP(3) IVPMVTL(3) IRPMVST(2) IRPMVYV(1) IRPMLPG(1) IRPMLPA(1) IVPMMPE(1) MPHWWYP(1) IRPMV 11 Phage display (common panning) 4 PMID:16229273 Anti-ochratoxin A monoclonal antibody 10G7 Ochratoxin A Not determined. Not determined. Ph.D.-7 phage display library (X7) 23 QLIMIRH(2) ITPRNRS(2) LQQKHLL(1) 3 Phage display (common panning) 3 PMID:16276928 Vibrio cholerae El Tor Typing Phage VP1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 24 RHKIQLRQNII(5) 1 Phage display (common panning) 3 PMID:16520318 Tumor necrosis factor ligand superfamily member 13B Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISAs were used to identify positive phages 25 WHWTYGWRPPAM(2) STYWSHYGQQPY(1) HWRYWHYPSETF(1) WHYSWFSPYRTY(1) TWWPHYHSLPRV(1) W-[SP]-H-Y 5 Phage display (common panning) 3 PMID:16864112 Acetylcholinesterase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 26 YHWDPLP(2) HPSYPRH(1) HPSDPRH(1) HHRDPCH(1) H-x(2)-D-P-x-H 4 Phage display (common panning) 4 PMID:17424851 Anti-aflatoxin B1 monoclonal antibody Aflatoxin B1 Not determined. Not determined. Ph.D.-7 phage display library (X7) 27 GSHHRHVHSPFV(3) HHPSHFSFSSPA(1) HHTHWPLSARTG(1) HHTHQASFMTRL(1) HHEHYKAVYTTH(1) HHYDTYWVWTTS(1) WAWPTHTHSPPP(1) YKFEYSEGHNLH(1) H-x(2)-H 8 Phage display (common panning) 4 PMID:16704119 Avian infectious bronchitis virus (strain M41) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) One linear peptide "GSHHRHVHSPFV" from the positive phages with the highest neutralization titer was synthesized and this peptide inhibited IBV infection in HeLa as well. 28 CLTPCRNKC(10) 3 Phage display (common panning) 4 PMID:16759988 Anti-arginine kinase monoclonal antibody 38G6 Arginine kinase Not determined. Not determined. CX7C T7 phage display library The peptide has an 83 29 AYQKWQTGA(2) EYQPHQTGL(1) PYENQQTGW(1) GYLPEQLGV(1) PYLFEQSGA(1) RYIALQLGL(1) NYRQDQLGM(1) SYRSLQTGW(1) YYAFGQIGL(1) AYQMYAIPA(1) Y-x(3)-Q-[LI]-G-[ST] 10 Phage display (common panning) 2 PMID:16273596 Anti-cholera toxin peptide 3 (CTP3) monoclonal antibody TE33 Cholera enterotoxin subunit B 1TET, Not determined. pVIII-9aa phage display library (X9) 30 CNQLFNTPPSC(5) CAQTFNSTTEC(1) CRQGLHTPPRC(1) CVRAPLLGPRC(1) CYGPLSSPLNC(1) QLFNTPP 5 Phage display (common panning) 2 PMID:16273596 Anti-cholera toxin peptide 3 (CTP3) monoclonal antibody TE33 Cholera enterotoxin subunit B 1TET, Not determined. pVIII-9aa.Cys phage display library (CX9C) 31 DWVAVKQSYF(2)[0.78/0.686, 5.43/5.08e-9] HLVAVIGSYR(1)[0.782, 7.28e-9] PSLFSWGFGS(1)[0.805, 8.54e-9] V-A-V-x(2)-S-Y 3 Phage display (common panning) 3 PMID:12917476 Shrimp white spot syndrome virus Not determined. Not determined. Not determined. X10 phage display library ELISA Phage clones amplified by panning were tested by ELISA for their ability to bind specifically to WSSV. HRP activity was estimated by measuring A405 using a universal microplate reader (EXL-800; Bio-tek). All samples were tested three times and the mean values are given. Besides the affinity (Kaff) constant of peptides for WSSV was determined according to the method of Beattyet using a solid-phase non-competitive enzyme immunoassay. HLVAVIGSYR had a high specificity and blocked virus infection, with the possible critical motif for virus inhibition being VAVXXSY. 32 VPWMEPAYQRPL[0.36] TLGAQYLPTRTA[0.17] MPPTDYCSLCIK[0.20] PPAHNSKSPMKR[0.29] MPYTNERNPSQP[0.49] TPQLTTGIYPPR[0.58] SLTMMSPPAASD[0.54] LKMPVSELLLKK[0.38] VPLPWHMYAPAR[0.85] 9 Phage display (subtractive panning) 3 PMID:12527505 Osteosarcoma cell line OS-732 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance at 490 nm was dertemined. Results demonstrated that only 9 clones specifically bound to osteosarcoma cells. Data shown were reproduced from the graph. Osteoblasts were used as the absorber cells for subtraction biopanning from phage display library. 33 SNFASITTPRPH WPTNPTTVPVPS LTSDTYFLPVPA SLHWPVSHPPPP SVSVGMKPSPRP WHSQRLSPVPPA SNQGGSPLPRSV SEPHLPFPVLPH LPLPAPSFHRTT YPLPHPMWSMLP TMTPPPTSVRGT TPLPTIRGDTGT GPPPHHRDYHGP YPAPIKVLLPNS SPYPMALFPLHN SPYPSWSTPAGR P-x-P 16 Phage display (common panning) 3 PMID:12480936 Virion infectivity factor Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 34 ENHSPVNIAHKL(2) ENHSPVNIDHKL(2) ENHSPVNIAHKV(1) EDHSPVNIDHKL(1) ENHYPLHAAHRI(1) ESHQHVHDLVFL(1) TSHHDSHGDHHV(1) PGHHDFVGLHHL(1) 8 Phage display (common panning) 3 PMID:14607870 Anti-ViCPS monoclonal antibody ATVi Capsular polysaccharide Vi antigen (ViCPS) Not determined. Not determined. Ph.D.-12 phage display library (X12) 35 TSHHDSHGLHRV(11) TSHHDSHGVHRV(2) ENHSPVNIAHKL(2) DNHSPVNIAHKL(2) TSHHDSHDLHRV(1) TSHHDYHGLHRV(1) ENHYPVNIAHKL(1) 7 Phage display (common panning) 4 PMID:14607870 Anti-ViCPS monoclonal antibody ATVi Capsular polysaccharide Vi antigen (ViCPS) Not determined. Not determined. Ph.D.-12 phage display library (X12) 36 ENHSPVNIAHKL(3) DNHSPVNIAHKL(1) QNHSPVYIAHKL(1) QNHSPVNIAHKI(1) FPIAALNNETSF(1) QNTDQATPHRML(1) QITDQVNVHHML(1) TNHLGLQSSHRF(1) 8 Phage display (common panning) 3 PMID:14607870 Polyclonal antibody PTS Capsular polysaccharide Vi antigen (ViCPS) Not determined. Not determined. Ph.D.-12 phage display library (X12) 37 TSHHDSHGLHRV(16) ENHSPVNIAHKL(2) 2 Phage display (common panning) 4 PMID:14607870 Polyclonal antibody PTS Capsular polysaccharide Vi antigen (ViCPS) Not determined. Not determined. Ph.D.-12 phage display library (X12) 38 CRELRYLRRAC(17) CPRPLRGVYMC(3) CPRSLRHLANC(2) CLLEKLRMRAC(1) 4 Phage display (common panning) 3 PMID:16361318 Anti-IL-4 monoclonal antibody 11B.11 Interleukin-4 Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) The paper suggests that the amino acid residues spanning from 79 to 86 (QRLFRAFR) on IL-4 are of the major binding site for 11B.11. 39 GQQQTPY(1) GLQQASV(1) VQETQRP(1) GLQATPA(1) VTQRLPL(1) TQDTPRT(1) QGRIPTS(1) QSNEPRR(1) IAHHQFP(1) IKTQALM(1) EKHQPER(1) Q-X-[PTS] 11 Phage display (common panning) 2 PMID:17075129 Protein-glutamine gamma-glutamyltransferase 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Database searches showed that several proteins contain peptides similar to the phage-selected sequences, and the N-terminal glutamine-rich domain of SWI1/SNF1-related chromatin remodeling proteins was chosen for detailed analysis. 40 QQQHFLV(1) QQLHLEA(1) QQLSIPP(1) QQSPLWH(1) QQWITVP(1) AQHQLEL(1) WQIPMQL(1) WQTPMNS(1) WQRPYPH(1) SQLWLLP(1) SQLTLLP(1) TQFNPRY(1) FLTPTQP(1) AWPQTSA(1) VPNVHQK(1) Q-X-[PTS] 15 Phage display (common panning) 3 PMID:17075129 Protein-glutamine gamma-glutamyltransferase 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Database searches showed that several proteins contain peptides similar to the phage-selected sequences, and the N-terminal glutamine-rich domain of SWI1/SNF1-related chromatin remodeling proteins was chosen for detailed analysis. 41 CDVPCFGWCVAWC(12) CEGVCRVSCFGWC(3) CDPPCFGWCQDAC(2) CQGGCRVQCFGFC(1) CTGACPVVCFGFC(1) ILCFGMC(1) CDVPCFGWCVSWC(1) CVCFGYCRYEC(1) CHILCFGYCGDLC(1) CEGKCVVSCFGFC(1) C-F-G-[WYF]-C 10 Phage display (common panning) 2-5 PMID:16813566 Plasminogen activator inhibitor 1 Vitronectin 1OC0, Not determined. X7+CX7C+CX10C+CX3CX3CX3C phage display library pool Phage and PAI-1 expressed from HT-1080 cells (100 nM) were incubated in MBS supplemented with BSA and glycerol for 1 h. Formed phage-PAI-1 complexes were captured on polystyrene-adsorbed monoclonal anti-PAI-1 antibody Mab-7 (first and fourth rounds of panning), Mab-5 (second and fifth rounds of panning) or Mab-3 (third round of panning). Individual phage from selection rounds 2-5 were tested for PAI-1-binding in a sandwich ELISA. Sequencing of positives identified a short disulfide-constrained loop CFG(W/Y/F)C as a well-defined consensus motif. A peptide with the sequence DVPCFGWCQDA (referred to as paionin-1 hereafter) was synthesized. The paionin-1 sequence is a chimaera of clones A (CDPPCFGWCQDAC) and B (CDVPCFGWCVAWC) devoid of the flanking cysteine residues, which might be not essential as indicated by the clone G (ILCFGMC). The paionin-1 peptide inhibited the PAI-1 binding of phage clone B, while a linear version paionin-1 with the two cysteine residues replaced by serines had no effect on binding. This suggested that the constraint imposed by the disulfide locked the CFGWC loop in a conformation required for binding or that the cysteines were of contact residues. The binding of paionin-1 to PAI-1 was evaluated further through competition experiments with vitronectin. The results showed that vitronectin significantly inhibited the binding of paionin-1 to PAI-1. Therfore, vitronectin was taken as the template of this set of mimotopes. 42 AARRPFPAPS(19) CFRQGCWVIT(3) SRFKVWWAAG(1) PARRPFPVTA(1) 4 Phage display (common panning) 4 PMID:16527822 Kallikrein-2 Not determined. Not determined. Not determined. X10 phage display library 43 FRTCLRSWACM(6) CYRMPTCMQRD(4) 2 Phage display (common panning) 4 PMID:16527822 Kallikrein-2 Not determined. Not determined. Not determined. X11 phage display library 44 TTSPLSQGSSYI(9) 1 Phage display (subtractive panning) 3 PMID:16369013 Listeria monocytogenes Cation-independent mannose-6-phosphate receptor Not determined. Not determined. Ph.D.-12 phage display library (X12) Panning was performed using Dynex Immunolon microtiter wells coated with L. monocytogenes cocktail (15 serotypes or strains, i.e. 1/2a, 1/2b, 1/2c, 3a, 3b, 3c, 4a, 4b, 4c, 4d, 4a, EGD and three EGD Mutant strains). Nonspecific phages were removed by absorbing the phage library sequentially on uncoated wells and wells coated with BSA. Subtractive selection was carried out by the sequential application of L. innocua cocktail and L. ivanovii cocktail to wells with washing between steps. Low-affinity L. monocytogenes-specific phage peptides were removed by rapid exposure and removal of L. monocytogenes cocktail. The wells were washed before incubation with L. monocytogenes cocktail (30 min) to elute specific high-affinity bound phages. TTSPLSQGSSYI is quite similar to the segment PLAQSGGSSYI in the human cation-independent mannose 6-phosphate receptor (M6PR), which is within the mannose 6-phosphate binding site of the M6PR. Binding of the labeled synthetic PLAQSGGSSYI peptide by Listeria spp. was confirmed by fluorescence spectrophotometry and FACS. Further evidence for binding of the receptor by L. monocytogenes and L. innocua was provided by affinity purification. M6PR is thus indentified as a novel receptor for binding and invasion of Listeria species. 45 HHWTYWLSGTVA FHWPRSWVTWQS SHWWWWDARGYD WHWQWTPWSIQP WHHPWWYPRPGV WHWHPLSWRYST YWPSKHWWWLAP H-W-x-W 7 Phage display (common panning) 3 PMID:16312021 Prostate-specific antigen Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After three biopannings, 36 clones were randomly isolated and their DNA was sequenced. The deduced amino acid sequences of the corresponding inserts identified 22 different sequences. When tested in an ELISA test for their reactivity, seven clones gave a positive signal with PSA, PSA-ACT but not with ACT, indicating that the recognition was directed against a region of PSA not covered by ACT. 46 VRWGSFAAWL(20) RLNWAWWLSY(5) IPVKWLLRWR(3) LRLQWRAWLA(2) PVRWRWASWL(1) 5 Phage display (common panning) 4 PMID:16524885 Macrophage receptor MARCO Not determined. Not determined. Not determined. X10 phage display library The VRWGSFAAWL phage-sMARCO interaction had significantly slower dissociation kinetics than that between sMARCO and lipopolysaccharide or lipoteichoic acid. Further work with this phage, and the second most enriched phage, displaying the peptide RLNWAWWLSY, demonstrated that both peptides bind to the SRCR domain of MARCO, and that they probably bind to the same site. 47 MDDRGLLLVKKKKHG(24) KKKRHDVEKHIPHVRAS(18) FDGVRKKSRGKSLEY(15) RRGDHDHDIFHWWVH(11) FLDERNYIKKKRHKL(5) SIEVLRGAMHVAPRR(1) GGWYDRKHRRPAPLS(1) KFFRKKSHYHSRTTS(1) 8 Phage display (common panning) 3 PMID:7649995 Heat shock cognate 71 kDa protein BAG family molecular chaperone regulator 1 1HX1, Not determined. X15 phage display library The peptides containing basic sequences with at least KK, KR or RR were enriched. A similar synthetic heptamer NIVRKKK had the highest affinity for Hsc70, and substitution analyses showed that hydrophobic residues followed by basic residues play important roles in maintaining this affnity. 48 CQFDLSTRRLKC(7) CQYNLSSRALKC(2) CVWQRWQKSYVC(1) CMWDRFSRWYKC(1) 4 Phage display (common panning) 3 PMID:16288119 Cetuximab Epidermal growth factor receptor 1YY9, Not determined. CL10 phage display library (CX10C) The in vitro biologic features of mimotope-induced antibodies are similar to those of the monoclonal antibody cetuximab. 49 CQMWAPQWGPDC(51) CKLYWADGEFTC(18) CVDYHYEGAITC(8) CKLYWADGELTC(3) CVDYHYEGTITC(1) 5 Phage display (common panning) 3 PMID:15210798 Trastuzumab Receptor tyrosine-protein kinase erbB-2 1N8Z, Not determined. CL10 phage display library (CX10C) The results indicate that the selected mimotopes are suitable for formulation of a breast cancer vaccine because the resulting Abs show similar biological features as trastuzumab. 50 SSTQVQHTLLQT KVTLHHPPITRS 2 Phage display (common panning) 3 PMID:15684661 Anti-omp40 monoclonal antibody Pg-ompA1 Outer membrane protein 40, Omp40 Not determined. Not determined. Ph.D.-12 phage display library (X12) The minimal epitope requirements of the MAb and the predicted amino acid sequences were identified in the region of 96IALDQTLGIP105 in 40-kDa OMP. 51 MMELGRV QVWPSSH VNPTYGN 3 Phage display (common panning) 3 PMID:15684661 Anti-omp40 monoclonal antibody Pg-ompA1 Outer membrane protein 40, Omp40 Not determined. Not determined. Ph.D.-7 phage display library (X7) The minimal epitope requirements of the MAb and the predicted amino acid sequences were identified in the region of 96IALDQTLGIP105 in 40-kDa OMP. 52 GDCFFGFLNSPWRVC RSSYWVYSPWRFISR SPWR 2 Phage display (common panning) 3 PMID:12855711 Anti-vWF monoclonal antibody 82D6A3 von Willebrand factor (vWF) 2ADF, Not determined. f3-15mer phage display library (X15) 53 CMTSPWRC CRTSPWRC CYRSPWRC SPWR 3 Phage display (common panning) 4 PMID:12855711 Anti-vWF monoclonal antibody 82D6A3 von Willebrand factor (vWF) 2ADF, Not determined. CX6C phage display library 54 ECTRWSNRSRCF(13) QCHTWSNRRSCL(13) RCHAWSNRKSCV(8) KCGRWSNRSSCT(8) KCEPDDPWPQCI(7) QCGRWSNRSYCS(4) ACTTWSNRSKCP(4) NCGKWTNRRTCL(3) CGRWFNRSDLHC(3) KCSRWTNRHLCD(2) KCTRWTNRHLCS(2) KCTRWTNRAHCP(2) TCHRWGNRTSCQ(2) SCHAWSNRRTCR(2) ACHEWSNRSTCT(2) ACKRNHRWGACV(2) CMKWSNRSSRWC(1) QCSKWVNRSRCA(1) CSKWHNRSKRHC(1) CSKWANRLVSIC(1) QCSRWSNRTSCT(1) QCHRWANRISCS(1) RCTQWTNRAYCP(1) ACTQWSNRHMCG(1) KCGPWSNRSSCT(1) TCHPFSNRSTCT(1) ECGSHAWGRRCK(1) W-x-N-R 27 Phage display (common panning) 3 PMID:12676786 Anti-coagulation factor VIII monoclonal antibody BO2C11 Coagulation factor VIII 1IQD, Not determined. X15+XCX8CX+X9+CX10C phage display library pool 55 LDSMHFPFHSRSFWP NLSCTHPLGSPPPAP GQICYYGRDAYLCFL CESSLCLMYSLGPPA QTPPCPIEHCPSFYQ QSTCLSHPLLCLSWN PNCWVGLTGAHSCFL THSVPVAYPWPDLNA SPLDYECISHATVCF YSTPSSILDTHPLYK TLPPPCLSSPSRCVN RTMHPSDEFLPLGMP GTGLVPLFDPRYRFL SSSRQEPYPLYPLFS HPKVGEGIDFTSIVP ATDLLAAYPLYSPSL VVPLGRCVSHPAICA GFPCLSVASACYGIT 18 Phage display (common panning) 3 PMID:16630634 Anti-spike glycoprotein monoclonal antibody 80R Spike glycoprotein 2GHW, Not determined. f3-15mer and f88-15mer phage display library pool 56 RSGGCVGGQYCLTPTH NDWPCLSHTTVCNGTQ ATMPCLSHPSVCKHLY PMHECLSAPSVCADNY TELACLSEAYICDRSN ETFTCISAPWTCVTWL EKMACLSTLDVCMENP NNMSCLSHETICGRNP LPFECISKREVCDTPM SVDDCRWNLNCEPPP SEVYCPRPDRCLRAP VQRDCRWTFSCATLI TPPRCSDQMYCSLSR THQFCPDPKHCLAQP RMPPCMNAGECPTIA DTPDCXGNEKCLEYA TSNFCPAGGPCSPHG NPRVCMNKWECEQAI GPPLGCLSLSCYDVA WNDYCTMNQCDTHN KPLHCGDTFCSLNQ YLEHCTMNECLNAR NGYHCLSEFCMPHP SMEECRLWLCPPYE YKPWCEMNKCKPLA VMPECLSRLCDFDM DDMPGCYPMCTLNK YDSYCIMNFCGHAA YTAADCPGLLYLCP NDVRCKLWLCPMPD NNWPCLNETCPTKG VQWPCLSKQCNDNI YQADCLMNRCPTAE SAPECHLYYCPEQA ANPVCRLWMCPPIV RQTEPCNLWFCPQV REPPCVQVHCSTAK PKEQPWSEFRPAGM ADCTLWFCPQTSN CLSATCDCTLCGP FPELTCWTCLASS PPAYSCLCPWAHM 42 Phage display (common panning) 3 PMID:16630634 Anti-spike glycoprotein monoclonal antibody 80R Spike glycoprotein 2GHW, Not determined. f88-Cys1 phage display library (X5CXCX5) 57 TCLWSDLRAQCI REKRWIFSDLTHTCI WSD 2 Phage display (common panning) 4 PMID:16940148 Anti-gp120 monoclonal antibody b12 Surface protein gp120 (SU) 2NY7, Not determined. LX8 and X15CX phage display library pool 58 HSFGDLSISPNSLTAWPGTL WLMRAAWPWDYT NLRSTSFFELWAKWP GEFPDWDNLPLLCEG GRPQWYLQFEDYWRS DSLRVDEHHEVMVPM DWVPFFGFFFSRLQLP NWPRWWEEFVDKHSS VRFHLAGWLPAVVSFVIFSDH DGLRSDGSWLARFVFNGSGFY GNFTASTASWGDELLALY FVWEEHVGFLMRN NWPRWEEFVDKHSS FAHEGSASFRLSSKVEDWVSR CVFYANVEEEVQCWL FCVRLMCSGLIPFFVLCCFFA FSRADFFPLSYSLSSVPSTAL FSPYPPDIVHTTAFSSFVNPVD SSFFYSALTPSFPSPYSQSSR RFFAFPMVCASFFLVAIAFFP PLPPSRFFITVCLTFLFSLSFF VRPCVASLLLFFCLLFLLLPS ICFPFNTRYCIFAMMVSSLVF QQAGSYPGCIDYYYCHASAIG CLPAYGCKSFREPF RHFVPLYLSVSYDGFSRGASI PIIHPHPPRIAMRVSISPFP TPTDSTVRGSSTMDGFLKSVY TASFWRSVSFPWVLSFLAFSH GAQVNRKCAWHPRHI VAKKLWVPQVSGSNF HWGVYR 32 Phage display (common panning) 3 PMID:16940148 Anti-gp120 monoclonal antibody b12 Surface protein gp120 (SU) 2NY7, Not determined. X15 and X21 phage display library pool 59 CWVWSDEYASLMGC CRVWSDATGGYISC CRVWSDEHSAQMAC CERVWSDEHENYTC CRVWSDEWNSYGDC CILWSDINDRFISC CLWSDLLSQYTKPC CQILWSDVRGKFEC CPWMWSDILDRAIC CMWSDYLNRC CMWSDLYDELVNC CLYSDLRGTYETPC CTQIYSDDLQRWTC CTLVYSDYVGDTIC CLWDDLYGTYAEGC CKLWNDESARYTGC CTLWDDELGRSTHC CLPAFLYDDREHPC CLYNVDTGTRSEYC WSD 19 Phage display (common panning) PMID:16940148 Anti-gp120 monoclonal antibody b12 Surface protein gp120 (SU) 2NY7, Not determined. CX12C phage display library 60 HQARPPWTV(3) QLTDAIRQH(3) HHRHSPDNL(3) SAWGRNWHN(3) LGQRTPKSY(2) LTVQARPVG(1) SFQWRPWHI(1) WHQNRGGQA(1) TIQQXPAL(1) SHKQTGPG(1) HWNQPRWSS(1) HQQLPTRWD(1) FHLGTWRPG(1) VQAHGWGRP(1) YHKRPVGTP(1) LVQRYDHRT(1) DWRTPLQQH(1) RSLNLQDFN(1) RFSVETQI(1) ASAEKAHRR(1) ATGQYRGHS(1) TRHDYRQVL(1) VLSYRWDHW(1) FVGYRPAGW(1) HYRTLGDHR(1) GKVHHRNSS(1) AQGGRNQHW(1) MRNVTERHW(1) TSMLRNTSS(1) KRNWMNHGF(1) HSNSRSAQV(1) SNGHSVRQM(1) TSDRLLTIK(1) 33 Phage display (common panning) 3 PMID:7592831 Cytochrome b-245 Neutrophil cytosol factor 1, NCF-1 1WLP, Not determined. J404-3 phage display library (X9) 61 MFSPWNDHF(3) RVRQFG(2) VRHDLLF(2) VRPARHWWK(2) DLTRQG(2) SSVHYM(2) KGAPWQEHE(2) STRVRWGAY(1) ATRVPRKHW(1) TRVMRE(1) TRPRWHKHA(1) KRHRRKTWG(1) LRVWNL(1) WHKHWTRPV(1) WHKTWRVKG(1) KVLPDVRAA(1) VRAVTL(1) VRPPGG(1) SAVRIL(1) ARLAVR(1) ARSPRQIHI(1) ILRQAN(1) NRHIRRL(1) VYGGSRQWG(1) STRVRRQL(1) ALRDLLYPD(1) KFDLLSPFS(1) LGDLLKDMY(1) WHDLGF(1) WDFSDLHW(1) DVSWDLITA(1) AARVDPLVK(1) VPARFDLDN(1) PGGDLY(1) TFDLTS(1) SSGPWKEHF(1) ESGAFA(1) FRIGHF(1) GSHFSIFS(1) PALPWAEHF(1) VHIMKL(1) PWSEHF(1) 42 Phage display (common panning) 3 PMID:7624379 Neutrophil cytosol factor 1 Cytochrome b-245 heavy chain Not determined. Not determined. f3-6mer and J404-3 phage display library pool 62 GGPPRV(1) FTKKIYGPP(1) YLDGGP(1) HGMKFPGPA(1) TQQRGP(1) LMVKPREKHW(1) KWHKNHPP(1) FTWPRW(1) AHKHWRPRT(1) 9 Phage display (common panning) 3 PMID:7624379 Neutrophil cytosol factor 1 Cytochrome b-245 light chain 1WLP, Not determined. f3-6mer and J404-3 phage display library pool 63 CPSSVRHSC(7) CSLRPLEIC(5) CKATNMHMC(2) CRATPEPGC(2) CSSANHQTC(2) CESNKPWYC(2) CPYKGPSEC(2) CILTDMPHC(2) CWKTPTGKC(1) CYLAHTAKC(1) CHSVAAKWC(1) CISSGRPLC(1) CNMSSLTIC(1) CKSLNDRHC(1) CQPRSILAC(1) CSTTRLQHC(1) CGRLPQGVC(1) CTWASKLMC(1) CQNNDSSMC(1) 19 Phage display (competitive panning) 3 PMID:12639821 CD8+ T cell line Monoclonal antibody 4B4 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Peptides were eluted from a T cell line by 4B4 mAb. 64 MTAPPRN(13) NTSSQRP(3) ACPYKGP(2) GPVNPLS(2) FTRVPAN(2) FSAYRIG(2) FSPPPNT(1) AARPPQT(1) AARMPGL(1) MAPHPLI(1) MSSPNHA(1) MMNSPPA(1) MTRSPPA(1) LSRTVPA(1) TASIWRA(1) SYSPSPP(1) QLSITRL(1) 17 Phage display (competitive panning) 3 PMID:12639821 CD8+ T cell line Monoclonal antibody 4B4 Not determined. Not determined. Ph.D.-7 phage display library (X7) Peptides were eluted from a T cell line by 4B4 mAb. 65 SLSSRQSP(8) CTSSRPSG(1) SGFGRLSD(1) RSQTRKSK(1) KKQGRDST(1) RKQKRRTE(1) PPSFRRSS(1) LPTGRATT(1) NTPTKLSP(1) RRPTKKNT(1) RGEKRSKS(1) MLLIRTWE(1) VTYARLCY(1) LSYRKLRF(1) GTRRREEH(1) DRKGRQQQ(1) RYPCRYGL(1) KEENRKNN(1) FHPSRHPP(1) IAREKGQQ(1) ICPPRLLQ(1) 21 Phage display (common panning) 2 PMID:9395536 Tryptase Fibrinogen alpha chain Not determined. Not determined. X4[RK]X3 phage display library 66 SLSSRQSP(12) 1 Phage display (common panning) 4 PMID:9395536 Tryptase Fibrinogen alpha chain Not determined. Not determined. X4[RK]X3 phage display library 67 WRAFLPRWHA(1)[0.22] WNRGWRWWMG(1)[0.18] GFFWKWRIGR(1)[0.16] HIRWKGHISW(1)[0.09] [HKR](2)-[HKRFYW](2)-H 4 Phage display (common panning) 3 PMID:9858597 Cryptococcus neoformans melanin Not determined. Not determined. Not determined. L100 phage display library (X10) ELISA Absorbance at 405 nm was measured with a Ceres 900HDi (Bio-Tek Instruments Inc., Winooski, Vt.). Melanin-binding peptides were characterized by a high proportion of positively charged and aromatic residues. 68 RWPKRHLSGH(2)[0.73] LHKLVRHGRW(1)[1.94] YERKFWHGRH(1)[1.67] YLRRHTHVFW(1)[1.65] KKHSHYWVRY(1)[1.63] EFGTRHMRHR(1)[1.47] YRHHAHGGRG(1)[1.24] RKKWHGWTRW(1)[1.18] PKWRHGYTRF(1)[1.12] RHGTVKHARH(1)[0.94] RRHWHPPVQI(1)[0.90] EAYKRRWHWP(1)[0.86] SRVPFRHYHH(1)[0.62] RRPEHTKARW(1)[0.44] [HKR](2)-[HKRFYW](2)-H 14 Phage display (common panning) 4 PMID:9858597 Cryptococcus neoformans melanin Not determined. Not determined. Not determined. L100 phage display library (X10) ELISA Absorbance at 405 nm was measured with a Ceres 900HDi (Bio-Tek Instruments Inc., Winooski, Vt.). Melanin-binding peptides were characterized by a high proportion of positively charged and aromatic residues. 69 CELYENVGMYC(18)[0.49, 10-25] 1 Phage display (common panning) 3 PMID:9360978 Growth factor receptor-bound protein 2 Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. CX9C phage display library ELISA,Surface plasmon resonance (SPR) The G1 phage is attached to the plate surface and GST-Grb2 domain is allowed to bind at 3 M of protein concentration. Binding was detected using anti-GST antibodies. Absorbance at 405 nm was measured. Data shown were reproduced from the graph. Besides, the IC50 values are determined by mixing peptide with recombinant Grb2 or Grb2 SH2 domains and then measuring the amount of binding at equilibrium to the immobilized SHC phosphopeptide, biotin-DDPSpYVNVQ. Synthetic G1 peptide blocks Grb2 SH2 domain association (IC50 10–25 μM) with the SHC phosphopeptide. In fact, the recombinant GST-Grb2 protein is target. 70 RSANAI[4700] RNAQVR[3200] RKASLS[1010] RSATRD[740] SGRARQ[NT] SKSGRS[NT] SSRNAD[NT] TARLRG[NT] TARSDN[NT] TSRMGT[NT] TSRQAQ[NT] TTRRNK[NT] TTSRRS[NT] WSGRSG[NT] AIKRSA[NT] GRRGNR[NT] GRSVNN[NT] HTRRMK[NT] ISTARM[NT] KAADVT[NT] KKRTND[NT] KMSARI[NT] KRRDVA[NT] KRVSKN[NT] KSADAA[NT] RAAAMV[NT] RAGNIR[NT] RAHRDN[NT] RARDDR[NT] RARHMV[NT] RARSPR[NT] RAVGHQ[NT] RAVVDS[NT] RGGKGP[NT] RGRSAV[NT] RGVDMN[NT] RGVKMH[NT] RHRSDI[NT] RKGQGG[NT] RKLHMN[NT] RKMDMG[NT] RKMDRS[NT] RKMRMG[NT] RKNQRV[NT] RKQRDS[NT] RKRVGA[NT] RKSKVV[NT] RKSTSS[NT] RKVGSL[NT] RKVPGS[NT] RKWISG[NT] RLATKA[NT] RMRKND[NT] RNAVEP[NT] RNDRLN[NT] RNGKSR[NT] RNMPLL[NT] RNTGSH[NT] RRMTMG[NT] RRRLNM[NT] RRTLDF[NT] RRAVSN[NT] RSAKVD[NT] RSAVVK[NT] RSDQFL[NT] RSDNPN[NT] RSERSL[NT] RSGDPG[NT] RSGNTT[NT] RSGNMG[NT] RSNGVG[NT] RSPDGM[NT] RSRRLP[NT] RSRVTS[NT] RSSHSS[NT] RSSOAA[NT] RSSSSH[NT] RSSSTV[NT] RSTDLG[NT] RSTNVE[NT] RSTRHK[NT] RSYTNS[NT] RTSPST[NT] RTSVNL[NT] SKRASI[NT] SQTCVR[NT] TERRVR[NT] TQRSTG[NT] TRRDRI[NT] VARNYK[NT] VSRRNM[NT] SGRSA 90 Phage display (common panning) PMID:9252355 Urokinase-type plasminogen activator Not determined. Not determined. Not determined. X6 fAFF1-tether C phage display library Dot blot,Binding assay The dot blot analysis of the digestion of all 100 substrate phages was performed by u-PA using a wide variety of stringencies of digestion (data not shown). Although this semiquantitative assay cannot provide kinetic constants, it can provide an accurate rank ordering of the lability of the substrate phage clones. Under the most stringent conditions examined, 11 of the 100 substrate phage, containing eight distinct randomized hexamer sequences (RKASLS, RNAQVR, RRAVSN, RSAKVD, RSANAI, RSATRD, RSAVVK and RSSSSH), proved to be particularly labile u-PA substrates. Besides, four peptides (RSANAI, RNAQVR, RSATRD and RKASLS) were chosen for detailed kinetic analysis and compared with the hydrolysis of a control peptide containing the P3–P4' sequence of plasminogen. All four of the selected peptides were substantially improved substrates for u-PA, by factors of 840–5300, compared with the control, plasminogen peptide (kcat/Km, 0.88 1/Ms). Data shown were kcat/Km (1/Ms). NT denotes not tested. 71 SVTVVPFRWYSCS[43000.0 ± 12600.0] SLDFNSFRWCSAL[15900.0 ± 4070.0] SSLECSFRWGPEH[3700.0 ± 580.0] SSIISHFRWGLCD[7.6 ± 2.4] 4 Phage display (common panning) 3 PMID:9346944 Melanocyte-stimulating hormone receptor, MSH-R Melanotropin alpha, Alpha-MSH Not determined. Not determined. S-(X)5-FRW-(X)4 phage display library Binding assay All peptides were synthesized as amides, and affinities to the hMC1 receptor were characterized in standard ligand binding assays with the insect cell. Ki (nm) values for the different peptides on the MC1 receptor are measured and shown. 72 CQDPICFCGADGACYCTSRNC(12/16)[1.22 ± 0.04] CAWHYRFCGAAHSADGACREVFLVC(2/16)[0.32 ± 0.02] 2 Phage display (common panning) 2-3 PMID:16423833 Polymeric immunoglobulin receptor Not determined. Not determined. Not determined. Cys6 phage display library (CX6C) ELISA Absorbance was read at 405 nm (Tecan Sunrise Microplate Reader; Tecan Austria Gesellschaft, Salzburg, Austria) with Magellan 3.0 software. 73 CVVWMGFQQVC(6/13)[1.89 ± 0.07] CWTSGARWRLC(1/13)[0.33 ± 0.01] 2 Phage display (common panning) 2-3 PMID:16423833 Polymeric immunoglobulin receptor Not determined. Not determined. Not determined. Cys9 phage display library (CX9C) Absorbance was read at 405 nm (Tecan Sunrise Microplate Reader; Tecan Austria Gesellschaft, Salzburg, Austria) with Magellan 3.0 software. This study implied that the C9A peptide (CVVWMGFQQVC) sequence may be exploited for pIgR-mediated epithelial transport without interfering with secretory immunity. 74 CVVWMGFQQVC(13/35)[1.26 ± 0.03] CALVSEAGCLVWAA(12/35)[0.70 ± 0.06] CIIVPHAYAWC(6/35)[0.31 ± 0.03] 3 Phage display (common panning) 1-2 PMID:16423833 Polymeric immunoglobulin receptor Not determined. Not determined. Not determined. Cys9 phage display library (CX9C) ELISA Absorbance was read at 405 nm (Tecan Sunrise Microplate Reader; Tecan Austria Gesellschaft, Salzburg, Austria) with Magellan 3.0 software. This study implied that the C9A peptide (CVVWMGFQQVC) sequence may be exploited for pIgR-mediated epithelial transport without interfering with secretory immunity. 75 CPYGAALHC CYGGATLLC CQPAVANTC CSYLNVMHC CTGPLPNRC CNPTPEKRC CKPSSPPFC 7 Phage display (common panning) PMID:16928679 Protein tonB Iron(3+)-hydroxamate-binding protein fhuD Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 76 LLADTTHHRPWT HWKHPWGAWDTL KVWSLEPPGPAA YSPPSPEPPRIK QDRGILVEPPRM DFDVSFLSARMR KLWELNPPQVRT SPAPTNNYTYRL TQPLGLLPSRHL QTALITIHHSLT YGNSLPPRLGPP LWAKLWVVPERA SANLSWRESWPT 13 Phage display (common panning) PMID:16928679 Protein tonB Iron(3+)-hydroxamate-binding protein fhuD Not determined. Not determined. Ph.D.-12 phage display library (X12) 77 CPAPERPQC CHASPAHNC CVISAASQC CQSFPRQLC CNRPSSWLC CTAENSSPC CKTSPAWIC CMTARTTSC CISPAQSSC CPAVPAKAC CHLAPAARC CKALMRTSC CKPLFHNTC CHHWAPTRC CHNMPAQTC 15 Phage display (common panning) PMID:16928679 Iron(3+)-hydroxamate-binding protein fhuD Protein tonB Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 78 LHTPWHLPAPEI KSLSRHDHIHHH YHSPPHTPPAPL SFVGLVELPQNL VSRHQSWHPHDL KTLTLPLSNTSK KIMRMPRLMTRN LHFPLDYPQALG WHSPWSTPPAPS LHWPLYTPPASP 10 Phage display (common panning) PMID:16928679 Iron(3+)-hydroxamate-binding protein fhuD Protein tonB Not determined. Not determined. Ph.D.-12 phage display library (X12) 79 PQNSKIPGPTFLDPH(0.100)[72.2 ± 32.8] SMEPALPDWWWKMFK(0.088)[17.7 ± 9.4, 4.2 ± 2.3] DKPTAFVSVYLKTAL(0.013)[NA] APRPGPWLWSNADSV(0.013)[NA] GVTDSSTSNLDMPHW(0.013)[NA] PKMTLQRSNIRPSMP(0.013)[NA] PQNSKIPGPTFLDPH(0.013)[NA] LYPLHTYTPLSLPLF(0.013)[NA] 8 Phage display (common panning) 4 PMID:15528216 Galectin-3, Gal-3 Thomsen--Friedenreich glycoantigen, TFAg Not determined. Not determined. f88-15mer phage display library (X15) Fluorescence titration Dissociation constant (Kd, nM) described the binding of peptides to galectin-3. The results demonstrate that carbohydrate-mediated, metastasis-associated tumor cell adhesion could be inhibited efficiently with short synthetic peptides which do not mimic naturally occurring glycoepitopes yet bind to the galectin-3 carbohydrate recognition domain with high affinity and specificity. 80 ANTPCGPYTHDCPVKR(0.800)[72.2 ± 32.8] AYTKCSRQWRTCMTTH(0.013)[5.7 ± 2.2] LTGTCLQYQSRCGNTR(0.013)[NA] NISRCTHPFMACGKQS(0.013)[NA] PRNICSRRDPTCWTTY(0.013)[NA] 5 Phage display (common panning) 4 PMID:15528216 Galectin-3, Gal-3 Thomsen--Friedenreich glycoantigen, TFAg Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) Fluorescence titration Dissociation constant (Kd, nM) described the binding of peptides to galectin-3. The results demonstrate that carbohydrate-mediated, metastasis-associated tumor cell adhesion could be inhibited efficiently with short synthetic peptides which do not mimic naturally occurring glycoepitopes yet bind to the galectin-3 carbohydrate recognition domain with high affinity and specificity. 81 YPYSMWKNLWPS(5) PWWFNKYLFPPI(1) ANSFWVNHPTIT(1) W-x-N-x(2)-P 3 Phage display (common panning) 2 PMID:15847166 Anti-CSFV monoclonal antibody 1B5 E(rns) glycoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) 82 SVLFDKNRGQEI(17) QTCCDKNQGIFM(2) QPYDKNRGPTLN(1) SLYDKNRGQFGS(1) VLDWKTRGKDLP(1) D-K-N-[RQ]-G 5 Phage display (common panning) 2 PMID:15847166 Anti-CSFV monoclonal antibody b4-22 E(rns) glycoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) 83 QACQNNDTCPRL(14) TCRNNDTCRAPD(1) KACSNNSTCFN(1) PQPYDKNRGPTLN(1) SLYDKNRGQFGS(1) SFFDKNSAHWSP(1) HSWDKNSSIWWP(1) TWWDKNSATVAW(1) YYNKNNASYHSS(1) AMGYNKNNDSLL(1) DLPRWPGWYKNR(1) C-x-N(2)-x-T-C, YDKN 11 Phage display (common panning) 2 PMID:15847166 Anti-CSFV monoclonal antibody 24/16 E(rns) glycoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) 84 TRTNPWPAL(8) TRTQPGRFP(7) T-R-T-x-P 2 Phage display (common panning) 3 PMID:15634921 Anti-HMW-MAA monoclonal antibody 225.28S High molecular weight melanoma-mssociated antigen, HMW-MAA Not determined. Not determined. X9 phage display library The mimotope vaccine was then generated by coupling the most suitable candidate mimotope to tetanus toxoid as an immunogenic carrier. Immunization of rabbits with this vaccine induced a specific humoral immune response directed toward the epitope recognized by the mAb 225.28S on the native HMW-MAA. 85 TPFHHQHSTGFT NPLHHEHATGWT SWFDNFLYPTHD SYFDYLYPPRPA TYWEDTLYAARV QTFFDTMYPPPL 6 Phage display (common panning) 3 PMID:15908434 Anti-TRAP monoclonal antibody Signal transduction protein TRAP Not determined. Not determined. Ph.D.-12 phage display library (X12) By sequence analysis, it was found that the TA21 (SWFDNFLYPTHD) is highly homologous to the C-terminal sequence of TRAP which is conserved among S. aureus and Staphylococcus epidermidis, suggesting that peptide TA21 may be a useful broad vaccine to protect from infection caused by various staphylococcal strains. 86 CVHSPNREC(8) CNGSPNFVC(3) CYDTTNPSC(2) CLHATSPIC(2) CFPTRTNNC(1) CLAAYPHSC(1) SPN 6 Phage display (common panning) 4 PMID:15659541 Metallo-beta-lactamase L1 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 87 CRLTGGKGVGC(32) CRRTNWQGAGC(6) CADPNSVRAMC(4) CADPNSVRAHC(4) CQLTGTHGAGC(2) CAAHYRVGPWC(2) [RQ]-[LR]-T-[GN]-x(2)-G-[AV]-G, A-D-P-N-S-V-R-A-[MH], A(2)-H-Y-R-V-G-P-W 6 Phage display (subtractive panning) 3 PMID:15384175 SiHa cells Not determined. Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) To eliminate clones that bind plastic, the library was preincubated on 10 cm tissue culture dishes for 1 h at room temperature (RT) in DMEM 10% v/v FCS. SiHa cells were grown on 10 cm tissue culture dishes until they reached an 80% confluent monolayer in the conditions mentioned above. These peptides may be useful for the design of drug or gene delivery vectors for the treatment of cervical cancer. 88 GPDLPAGDS(5) GPAYSKDIR(2) GPHLPNGDA(1) GPSLKHTNM(1) GPALELESR(1) 5 Phage display (common panning) 5 PMID:15845542 Peptidyl-prolyl cis-trans isomerase A, PPIase A Not determined. Not determined. Not determined. pVIII-9aa phage display library (X9) The peptide FGPDLPAGD showed inhibition of the isomerase reaction and NMR chemical shift mapping experiments highlight the CypA interaction epitope. 89 FHLQSYH(28) 1 Phage display (common panning) 3 PMID:15528296 Anti-PSA monoclonal antibody PSA-C1 PSA-α2M complexes Not determined. Not determined. Ph.D.-7 phage display library (X7) 90 DLVRVIQ(32) 1 Phage display (common panning) 3 PMID:15528296 Anti-PSA monoclonal antibody PSA-C2 PSA-α2M complexes Not determined. Not determined. Ph.D.-7 phage display library (X7) 91 TPSASIP(10) 1 Phage display (common panning) 3 PMID:15528296 Anti-PSA monoclonal antibody PSA-C4 PSA-α2M complexes Not determined. Not determined. Ph.D.-7 phage display library (X7) 92 FPWSQSHHQASL AYPLSLISRAPP TSSLPKLPFAPE LPYFSLSPDPFA 4 Phage display (common panning) 2 PMID:15126333 DNA repair protein XRCC3 DNA repair protein RAD51 homolog 3, R51H3 Not determined. Not determined. Ph.D.-12 phage display library (X12) 93 YKPSETIRM(2)[0.384, 72.5%] RRPRLLDTA(1)[0.479, 65.7%] 2 Phage display (common panning) 1 PMID:12657725 Anti-gastric cancer monoclonal antibody MG7 Gastric cancer-associated antigen Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA The affinities of phage particles to mAb MG7 were measuered in ELISA inhibition sssay. The OD450 was measured. The inhibition rate was calculated as inhibition rate = 1 - Test OD450/MG7 OD450. 94 CPLSWPFPVFC(3)[0.288, 79.4%] CRFAPLSAVRC(2)[0.271, 80.6%] CHGPRMISFWC(1)[0.366, 73.8%] CNAIYARNAQC(1)[0.284, 79.7%] CTCHLRVYAQC(1)[0.386, 72.4%] CPLGSNHNLPC(1)[0.312, 77.7%] CGSAVRRPSRC(1)[0.381, 72.7%] CSLDLHPLRSC(1)[0.316, 77.4%] CRMCKGCYTSC(1)[0.288, 79.4%] CSWAPVYARNC(1)[0.268, 80.8%] 10 Phage display (common panning) 1 PMID:12657725 Anti-gastric cancer monoclonal antibody MG7 Gastric cancer-associated antigen Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA The affinities of phage particles to mAb MG7 were measuered in ELISA inhibition sssay. The OD450 was measured. The inhibition rate was calculated as inhibition rate = 1 - Test OD450/MG7 OD450. 95 YHWYGYTPQNVI QIQVFHRWMTGP 2 Phage display (common panning) 3 PMID:16319141 Epidermal growth factor receptor Pro-epidermal growth factor, EGF 1IVO,1NQL,3NJP, Not determined. Ph.D.-12 phage display library (X12) Competitive binding assay and Scatchard analysis revealed that GE11 peptide (YHWYGYTPQNVI) bound specifically and efficiently to EGFR with a dissociation constant of ~22 nM, but with much lower mitogenic activity than with EGF. 96 QCTGPNVATNCR(6) QCTGPNFATNCR(1) TCNGPSVYMNCL(1) 3 Phage display (common panning) PMID:15905554 Anti-HMW-MAA monoclonal antibody 763.74 High molecular weight melanoma-mssociated antigen, HMW-MAA Not determined. Not determined. LX-8 phage display library (XCX8CX) 97 TCRLPFQNVACH(2) SCLLPFQNIFCS(2) 2 Phage display (common panning) PMID:15905554 Anti-HMW-MAA monoclonal antibody GH786 High molecular weight melanoma-mssociated antigen, HMW-MAA Not determined. Not determined. LX-8 phage display library (XCX8CX) 98 NQLPQYMGPAPAYMR(6) 1 Phage display (common panning) PMID:15905554 Anti-HMW-MAA monoclonal antibody GH786 High molecular weight melanoma-mssociated antigen, HMW-MAA Not determined. Not determined. X15 phage display library 99 LLGPYELWELSH(25) HPRPYHHTLPLT(1) 2 Phage display (subtractive panning) 3 PMID:15536075 Trastuzumab Receptor tyrosine-protein kinase erbB-2 1N8Z, Not determined. Ph.D.-12 phage display library (X12) Phages were preabsorbed on beads containing immobilized human normal IgG to remove any phages that were not specifically reactive with trastuzumab. According to the paper, peptide LLGPYELWELSH has potential for being developed as a HER-2 vaccine for biotherapy of cancer with HER-2 overexpression. 100 QDQRTFT VPATLFR LVTHTPI PHLHPPR MSLHHSH HAIYPRH ALRTASS 7 Phage display (common panning) 3 PMID:15812883 Protein P Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 101 FHLPYNHNWFAL(19) 1 Phage display (subtractive panning) 3 PMID:15618169 Anti-type 8 PS monoclonal antibody NAD Type 8 capsular polysaccharide Not determined. Not determined. Ph.D.-12 phage display library (X12) Between panning rounds 1 and 2, a subtractive panning was performed on a blank, blocked polystyrene plate to remove nonspecific plastic binders. Inhibition studies showed that FHLPYNHNWFAL is a mimetic of type 8 PS. 102 HPTLLRI(5) SLPTLTL(1) PVQMPWL(1) HAIYPRHH(1) HWGMWSY(1) ITNDFKQ(1) 6 Phage display (common panning) 4 PMID:15162530 Anti-NE2 monclonal antibody 8C11 Capsid protein Not determined. Not determined. Ph.D.-7 phage display library (X7) 103 SILPYPY(3) ALWGPTS(1) LGTLPSA(1) SPKMPIL(1) FSYLPSH(1) STLFMNF(1) SPPNVR(1) SAHVLST(1) 8 Phage display (common panning) 4 PMID:15162530 Anti-NE2 monclonal antibody 8H3 Capsid protein Not determined. Not determined. Ph.D.-7 phage display library (X7) 104 MHGPTKNQMWHT(7)[0.687/0.814] 1 Phage display (common panning) 3/4 PMID:15207087 Anti-hantaan virus monoclonal antibody F3 Hantaan virus Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Phage clones F3-8A with the sequence of MHGPTKNQMWHT and F3-8B harboring the sequence of MHGPTKNQMWHT were tested by ELISA for their binding specificity to anti-hantaan virus monoclonal antibody F3. The absorbance was determined as A450/A630 nm. Firstly the mAb F3 (100 mg/L) was incubated with phages. When the titer of mAb diluted 400 times, the absorbances were measured and shown. 105 MHRHAHPMYQPP(3)[0.814] MHRHYHPWLTPS(2)[NT] FHRTPWLQFSLP(2)[0.887] WPKHFHIPQWWT(1)[1.134] 4 Phage display (common panning) 3/4 PMID:15207087 Anti-hantaan virus monoclonal antibody B11 Hantaan virus Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Selected phage clones were tested by ELISA for their binding specificity to anti-hantaan virus monoclonal antibody B11. The absorbance was determined as A450/A630 nm. Firstly the mAb B11 (100 mg/L) was incubated with phages. When the titer of mAb diluted 400 times, the absorbances were measured and shown. NT denotes not tested. 106 CLPWSDGPC(5) CLPWGTGPC(1) 2 Phage display (common panning) 3 PMID:15564686 Anti-gibberellin monoclonal antibody 8E9 Gibberellin 3-beta-dioxygenase 1 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 107 CTPEQQFTC(8) CTKSPPLQC(4) CTPSALASC(3) CSWPNTSNC(2) CHATHTNYC(1) CTSLLRGQC(1) CPPMPNTTC(1) 7 Phage display (common panning) 3 PMID:17037189 Anti-SARS-CoV spike glycoprotein monoclonal antibody 2C5 Spike glycoprotein Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The results demonstrated that TPEQQFT is a mimic epitope peptide containing neutralizing MAb 2C5. 108 ASYPPRL ISYHDLR SLSPGLP STSFTPH MADRIGT FAGVPSW LNNQFWY TPGQYWA YTRPLPT FSAPVRY 10 Phage display (competitive panning) 7 PMID:16867257 Anti-IFN-alpha-2b polyclonal antibody Interferon alpha-2 Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted for IFN- Twenty-three positive clones were obtained after seven rounds of selection. Ten clones were randomly picked from the positive clones and were sequenced. The corresponding amino acid sequences suggested 3 groups homologous to the 3 domains of IFN-α2b, defined by residues 24–41, 43–49, and 148–158 of IFN-α2b. 109 NMFGSLTSHVTA(4) QVNQWSPLVNIR(3) QLYNNRSLFPAW(2) NGNHRTLSAHAH(2) LGNQGLSLTLRL(1) VNPHHMQLPKLH(1) DENLSLRPLFPK(1) QAENKIDVHLPI(1) N-x(3,5)-L 8 Phage display (common panning) 3 PMID:16186382 Glucokinase regulatory protein Glucokinase Not determined. Not determined. Ph.D.-12 phage display library (X12) 110 CSERSMNFC(18) CYGLPHKFC(12) CPSGAARAC(9) CLQHKSMPC(3) CVKSMVTHC(3) CSQRSMNFC(2) CQPLRHHQC(2) CLPHKSMPC(1) CPSGTARAC(1) CKQRPAWLC(1) CIPMNAPWC(1) CSLPFARNC(1) CGPARLSFC(1) CMGLPLRFC(1) [RK]-S-M, L-[PQ]-H-K, P-S-G-[AT]-A-R-A 14 Phage display (common panning) 3 PMID:15506167 Human airway epithelial cell 1HAEo-cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 111 KGSHEALHRHIR[0.556, 0.953] KPHSTHLPAHQP[0.255, 0.795] HTIHSKQSHKNS[1.920, 1.206] HSKLNNRHHALL[0.803, 1.112] TPHSKPHFHKLP[0.193, 0.876] HSRHNHYLHPWA[0.477, 0.974] HSKHTNHPPVGS[0.260, 0.849] MPKHIHRPHNIN[1.184, 1.007] WTVLPHEGGLSS[0.473, 1.381] LPSRHAAHPHVR[0.351, 0.906] KSLSRHDHIHHH[0.577, 0.775] 11 Phage display (subtractive panning) 3 PMID:15041534 Anti-hTERT polyclonal antibody Telomerase reverse transcriptase Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding of anti-hTERT polyclonal antibody with selected phages was quantified by both direct ELISA and sandwich ELISA methods. The absorbance was measured at 490 nm. Data shown were reproduced from the graph. In each round, a subtractive panning was performed on normal mouse IgG. After 3 rounds of screening, 13 of 24 randomly selected phage clones were identified as positive clones that could specifically bind to anti-hTERT anibody but not to normal mouse IgG. 112 WCRGGWC(4)[++] CLCAEGWC(4)[++] CHPGTGWC(3)[++] WCKSGWC(2)[++] WCVKGWC(2)[NT] SYCREGWC(1)[+] CGCRDGWC(1)[+] CLCHGGWC(1)[+] CRDGWCYS(1)[NS] CVAILKDC(1)[-] 10 Phage display (common panning) 3 PMID:14764731 Anti-CD47 monoclonal antibody C5D5 Leukocyte surface antigen CD47 Not determined. Not determined. CL6 phage display library (CX6C) ELISA ELISA results quantifying binding of such phage to mAb C5D5 are also shown with OD values ++ (>1.0), + (<1.0), - (background), NT (not tested) and NS (not significant). 113 CGWRNSFGQSLC(32)[++] CERVIGTGWVRC(6)[++] CHRVPGHGWVRC(5)[++] CVPVCREGWCGC(3)[NT] CYKSMDGWVVPC(3)[++] CVENVDGWTVPC(2)[++] CSWQHQDGWVWC(1)[++] CRVPETGWVKC(1)[NT] CRLMLNGWVVPC(1)[NT] CCRDGWCHHDWC(1)[++] CCREGWCGDGLC(1)[NT] CGWRNALQVVC(1)[NT] CGWRNLEGGSVC(1)[NT] CGWRDDSGQSMC(1)[NT] CRRVIGRVGCGC(1)[++] 15 Phage display (common panning) 3 PMID:14764731 Anti-CD47 monoclonal antibody C5D5 Leukocyte surface antigen CD47 Not determined. Not determined. CL10 phage display library (CX10C) ELISA ELISA results quantifying binding of such phage to mAb C5D5 are also shown with OD values ++ (>1.0), + (<1.0), - (background), and NT (not tested). Studies indicated that peptide CERVIGTGWVRC specifically blocked CD47 and SIRPα binding in a dose-dependent fashion. Protein binding assays using SIRPα domain-specific recombinant proteins demonstrated that this peptide directly bound to the distal-most Ig loop of SIRPα, the same loop where CD47 binds. 114 WLHLHSRPLPSTPHD(17)[220] AGDRPLPPLPYNPKS(8)[ND] LALARPLPVPPWRQI(2)[ND] TGPRPLPLPPLRSMS(2)[180] SFRPLPPLPQFLPMY(2)[185] STLMKISNRPLPAAS(2)[>10,000] RPGDPLRTPIAGDT(2)[>10,000] HSHFHPRPLPPLPVR(1)[417] FVGDPLPYIPHMHWF(1)[>10,000] R-P-L-P-x(2)-P 9 Phage display (common panning) 4 PMID:7929055 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. X15 phage display library Binding assay Individual peptides were immobilized via their amino termini to polystyrene plates. A constant input of 32P-labeled GST/PKA/Src SH3 (typically 1-3 nM), with varying amounts of unlabeled GST/PKA/Src SH3 (0-10 μM), was incubated with the immobilized peptide. Scatchard analysis of binding data was performed using the EBDA and modified LIGAND programs (Biosoft). Kd (nM) values were determined by Scatchard analysis of equilibrium binding data. ND, not determined. In fact, GST/PKA/SrcSH3 fusion protein is the target. 115 RSLPPIPG(10) 1 Phage display (common panning) 8 PMID:7988556 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. GX6G phage display library A point mutation in a flanking codon (GGG-AGG) had occurred, converting glycine to arginine. 116 SLARPL(1) TSMRPL(1) KSERPL(1) TLGRPL(1) SIARPL(1) APRIPL(1) LHRRAL(1) YDHRSL(1) LRQRPL(1) PASRPL(1) ARDRPL(1) PRSRPL(1) FVSRPL(1) NKGRSL(1) NRLRPL(1) LANREL(1) PTRRPL(1) x(3)-R-P-L 17 Phage display (common panning) 3 PMID:7988556 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. X6PPIP phage display library 117 RLLKPL(2) IQHRLL(1) QSRRSL(1) AKRAPL(1) TGGRPL(1) HHIRPL(1) SYPRPL(1) HSTRAL(1) x(3)-R-P-L 8 Phage display (common panning) 3 PMID:7988556 Fyn SH3 domain of tyrosine-protein kinase Fyn Not determined. Not determined. Not determined. X6PPIP phage display library 118 PTHRIL(1) RNFLIL(1) RVSYNL(1) PVLHSL(1) RYLRPL(1) RSSRPL(1) VNKRPL(1) RSNRPL(1) RAKRPL(1) SHRHPL(1) R-x(2)-R-P-L 10 Phage display (common panning) 3 PMID:7988556 Lyn SH3 domain of tyrosine-protein kinase Lyn Not determined. Not determined. Not determined. X6PPIP phage display library 119 ASTRPL(2) RLFRPL(1) RPQRPL(1) RFKRPL(1) RAKRPL(1) RPYRPL(1) RIPRPL(1) RSLRPL(1) R-x(2)-R-P-L 8 Phage display (common panning) 3 PMID:7988556 PI3K SH3 domain of PI3-kinase subunit p85-alpha Not determined. Not determined. Not determined. X6PPIP phage display library 120 PPPYPP(5) APPYPP(2) PPAYPP(2) APSYPP(2) APHYPP(1) PPPYHP(1) PPSYPP(1) APNYPP(1) APSYSP(1) PPHYPP(1) P(3)-Y-P(2) 10 Phage display (common panning) 3 PMID:7988556 Abl SH3 domain of tyrosine-protein kinase ABL1 Not determined. Not determined. Not determined. X6PPIP phage display library 121 PPLPPP(3) PPLPGS(2) PPLPRP(1) PPLPSP(1) PPVPPP(1) PPLPNG(1) P(2)-L-P-x-P 6 Phage display (common panning) 3 PMID:7988556 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. RSLRPLX6 phage display library 122 PPLPFD(1) PPPPFP(1) PPLPVP(1) PPLPWT(1) PPLPPL(1) PPIPGR(1) PPIPPP(1) PPLPAP(1) PPIPLG(1) PPIPDS(1) P(2)-[IL]-P-x(2) 10 Phage display (common panning) 3 PMID:7988556 Fyn SH3 domain of tyrosine-protein kinase Fyn Not determined. Not determined. Not determined. RSLRPLX6 phage display library 123 PPLPLR(2) PPLPAP(1) PPLPSP(1) PPLPLP(1) PPLPFP(1) PPLPPP(1) PPLPLL(1) PPLPQL(1) PPLPSA(1) P(2)-L-P-x-P 9 Phage display (common panning) 3 PMID:7988556 Lyn SH3 domain of tyrosine-protein kinase Lyn Not determined. Not determined. Not determined. RSLRPLX6 phage display library 124 PPLPPP(5) MVSLVP(1) LPFGPP(1) PPTPV(1) P(2)-L-P(3) 4 Phage display (common panning) 3 PMID:7988556 PI3K SH3 domain of PI3-kinase subunit p85-alpha Not determined. Not determined. Not determined. RSLRPLX6 phage display library 125 PPIPVR(1) PPVPYA(1) PPIPIL(1) PPVPYP(1) PPVPRS(1) PPVPGA(1) PPIPRF(1) PDIPLL(1) P(2)-[IV]-P-x(2) 8 Phage display (common panning) 3 PMID:7988556 Abl SH3 domain of tyrosine-protein kinase ABL1 Not determined. Not determined. Not determined. PPPYPPX6 phage display library 126 VEPPPGWG(2) LSPPPPGW(1) SPPPPGWG(1) VEPPPGWV(1) VDPPPGWG(1) VPPPPGWQ(1) EMPPPPGW(1) MAPPPGWS(1) LEPPPGFV(1) IDPPPGFM(1) IEPPPGWS(1) GGPPPPGW(1) GGPPPPGY(1) GQPPPGWA(1) GSPPPGWY(1) GGPPPGWQ(1) GAPPPGWQ(1) TGPPPGWG(1) GGPPPGWG(1) GTPPPGWG(1) GGPPPGWE(1) GHPPPGWE(1) HLPPPGWE(1) WGPPPGWS(1) AGPPPGWS(1) GGPPPGWS(1) EGPPPGWE(1) EGPPPGYE(1) EGPPPGWW(1) AGPPPGMP(1) LGPPPGMP(1) AIPPPGAP(1) EGPPPGMV(1) SGPPPGMM(1) GGPPPGMQ(1) TGPPPGYA(1) TGPPPGFA(1) GGPPPGYA(1) SWPPPGYQ(1) P(2)-G-[WFYML] 39 Phage display (common panning) 3 PMID:16000308 GYF domain of protein CD2BP2 Not determined. Not determined. Not determined. RKRSHRXXPPPXXXVQ phage display library 127 VGPPPGGR VGPPPGYG AGPPPGWG GGPPPGWS XEPPPGWE QPPPGWE ADPPPGWS SSPPPVGW LGPPPTWV P(2)-G-[WFYML] 9 Phage display (common panning) 6 PMID:16000308 GYF domain of protein CD2BP2 Not determined. Not determined. Not determined. RKRSHRXXPPPXXXVQ phage display library 128 KYPPGFTAP PPGYKKKLT NKGHPPPGW NKGHPPPAG GPPPGELSR GPPPGWLGR RQSAPPPGL KSTAPPPGW NPPPGHRAQ GPPPELAGA GRPPGSPAR P(2)-G-[WFYML] 11 Phage display (common panning) 6 PMID:16000308 GYF domain of protein CD2BP2 Not determined. Not determined. Not determined. X9 phage display library 129 GRPPPGFS ARPPPGWE GIPPPGLT GVPPPGYE GHPPPGME GSPPPGSE GWPPPGLD ESPPPGFP HGPPPGFE VAPPPGIE MVPPPGLE SGPPPGID SGPPPGLP VGPPPGLE VGPPPGLM EGPPPGLL AAPPPGLV EGPPPPGY SGPPPGIK P(2)-G-[FILMW] 19 Phage display (common panning) 6 PMID:16120600 GYF domain of protein PERQ2 Not determined. Not determined. Not determined. RKRSHRXXPPPXXXVQ phage display library 130 XGPPPGTW(2) ELPPPGTW(1) ETPPPGXW(1) SGPPPGMW(1) ATPPPGLC(1) GAPPPGAE(1) P(2)-G-[ILMY] 6 Phage display (common panning) 6 PMID:16120600 GYF domain of protein SMY2 Not determined. Not determined. Not determined. RKRSHRXXPPPXXXVQ phage display library 131 ESPPPGMX DSPPPGML QAPPPGLM DYPPPGLY VEPPPGLW PGPPPGLG NSPPPGLT AHPPPGWE HEPPPGLA P(2)-G-[ILMY] 9 Phage display (common panning) 6 PMID:16120600 GYF domain of protein SYH1 Not determined. Not determined. Not determined. RKRSHRXXPPPXXXVQ phage display library 132 PGPPPGLE PGPPPGLA EGPPPGLM ISPPPGME LTPPPGMD VEPPPGLD GEPPPGLV MEPPPGLW LGPPPGLG XGPPPQMX P(2)-G-[ILMY] 10 Phage display (common panning) 6 PMID:16120600 GYF domain of protein PR-SYH1 Not determined. Not determined. Not determined. RKRSHRXXPPPXXXVQ phage display library 133 APPGLRLQS(2) PPGLRPEHQ(1) PPGLSPLIR(1) PPGLPIATG(1) AQPPPSLQT(1) IPPGFSSIR(1) SKPPGFNEP(1) KHPPGMREP(1) FGPPGLDRP(1) ASPPGLDEP(1) RAPPGISSA(1) NGSPPGLSR(1) RSLPPGLSP(1) KPPPPGWPR(1) KNQRPPGSS(1) P(2)-G-[FILMW] 15 Phage display (common panning) 6 PMID:16120600 GYF domain of protein PERQ2 Not determined. Not determined. Not determined. X9 phage display library 134 GIPPGYFSP(4) PPGLNHSPR(3) APPPGKWEQ(1) XXPPGKWKP(1) PPPGVWRNE(1) GQPPGIWDL(1) TRPPGIPPR(1) LKSPPGLTR(1) CPPGLCEKL(1) RPPPGNLRA(1) P(2)-G-[ILMY] 10 Phage display (common panning) 6 PMID:16120600 GYF domain of protein SMY2 Not determined. Not determined. Not determined. X9 phage display library 135 PPGLNHSPR(3) SRPPGIIPL(1) RTPPGMLRL(1) GIPPGYFSP(1) HPPPGLFPP(1) LSRPPGLHW(1) PPGLHKERK(1) AFPPGLRAP(1) VMHPPGLRS(1) PARPPGISQ(1) PPPGLSTAP(1) LPPGSQDA(1) P(2)-G-[ILMY] 12 Phage display (common panning) 6 PMID:16120600 GYF domain of protein SYH1 Not determined. Not determined. Not determined. X9 phage display library 136 HPPPGIGPR(2) GRPPGLLRA(1) NGSPPGLXR(1) IPPPGITAR(1) SPPPGLHPP(1) RPPGLHVPE(1) RSPPGLSSP(1) NHPPGIHAM(1) P(2)-G-[ILMY] 8 Phage display (common panning) 6 PMID:16120600 GYF domain of protein PR-SYH1 Not determined. Not determined. Not determined. X9 phage display library 137 RTPPGFSRP(2) KYPPGVPPQ(1) GFPPGFGRR(1) KAPPGFGSP(1) RKPPGFGVR(1) RIPPGFEGR(1) SIPPGFGPW(1) RRPPRIETR(1) PPGFGPTQL(1) PFGPPGFQK(1) RKGPPPGFG(1) PPRVRQPNR(1) P(2)-G-F 12 Phage display (common panning) 6 PMID:16120600 GYF domain of protein GYN4 Not determined. Not determined. Not determined. X9 phage display library 138 WHDDNPFFFEAP(4) SLPPWYSPFKLD(2) KHYPEVGPNGFI(2) SHMPLANQYQWA(1) NHVQAWEQFWDS(1) DHSPFYLSTAHY(1) LHAYNPFLLSWP(1) RHFPNDYNPFLL(1) WHEAQPSPGLLR(1) DHSSADLINPVA(1) GWYSPFQLGWPQ(1) 11 Phage display (common panning) 6 PMID:17301381 Immunoglobulin fractions obtained from vitiligo patients Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) SHMPLANQYQWA and NHVQAWEQFWDS showed distinct positive reactivity with the patient’s sera compared with controls. 139 PRPEWRVHW PKAGGARPTW RRPTMMKHT PPGGTRPTGIQF PPGKLPSYRPYM MGVGAKRPG PPGTPPNRNS EVPPGV PPGDTVYRPIHW LERTRQPLPGPP PPGWRAPRL ATSGDAYWS MDAYDKISK GADMDRRIS WYDAKMRHS RRQEVTYSW VRYYDDYHR QLELDAQRG GFPWQKAYM HVDWRNSWR LETPPNRNS DHRLGSRNS GHLSSRNFV SQHWLSMWS LGQVEYMRR NGYRKQDGE DYFVNSRHN SFLVQTAKQ KLPSYRPYM LPWRMSGHV WSWWRNTHE THMWRN APVAGVRHN DWERFSWPY RFDRAGSRD ARWDFLRS RAEAQYSVV 37 Phage display (common panning) 3 PMID:8663333 Neutrophil cytosol factor 2, NCF-2 Neutrophil cytosol factor 1, NCF-1 1K4U, Not determined. J404-3 phage display library (X9) 140 WGANSPL(1) FGQNXYL(1) FGSNVHF(1) WGQNSFP(1) MWGENTP(1) APHWGQN(1) [WF]-G-[EQS]-N 6 Phage display (common panning) 3 PMID:9973494 Anti-lactotransferrin monoclonal antibody mAbRWL1 Lactotransferrin Not determined. Not determined. Ph.D.-7 phage display library (X7) 141 WGSNVYNSPFHS(5) SVSVGMKPSPRP(5) WGENRSNXTXPG(1) WGQNDSLPXAXK(1) FGENRSITTWPX(1) WGENNYTSQIRP(1) WGSNISXDLAQQ(1) WGENQITXRAPI(1) FGENNSIXGHRV(1) HSISLGATPTLP(1) VSAERXSRPTSP(1) [WF]-G-[EQS]-N 11 Phage display (common panning) 3 PMID:9973494 Anti-lactotransferrin monoclonal antibody mAbRWL1 Lactotransferrin Not determined. Not determined. Ph.D.-12 phage display library (X12) 142 CWGSNHAQC(1) CFGSNQSQC(1) CFGQNAPRC(1) CWGQNIKAC(1) CWGQNTSMC(1) CWGSNNQNC(1) CFGENTGYC(1) CWGANVPSC(1) CFGENNNMC(1) CPRSENGVC(1) [WF]-G-[EQS]-N 10 Phage display (common panning) 3 PMID:9973494 Anti-lactotransferrin monoclonal antibody mAbRWL1 Lactotransferrin Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 143 WGSNPAM(2) FGENRPA(1) WGSNVTS(1) WGTNSEV(1) WGANTNL(1) PYGLLTP(1) [WF]-G-[EQS]-N 6 Phage display (common panning) 3 PMID:9973494 Anti-lactotransferrin monoclonal antibody mAbRWL2 Lactotransferrin Not determined. Not determined. Ph.D.-7 phage display library (X7) 144 SVHMPVWGQNTS(3) WGENFIQVPVSE(1) YSVPPWGSNVMY(1) FGQNRANPQLPP(1) WGSNPPSATAPP(1) [WF]-G-[EQS]-N 5 Phage display (common panning) 3 PMID:9973494 Anti-lactotransferrin monoclonal antibody mAbRWL2 Lactotransferrin Not determined. Not determined. Ph.D.-12 phage display library (X12) 145 CWGENNYMC(1) CWGQNNWTC(1) CWGANPGLC(1) CFGSNDPYC(1) CWGSNAEHC(1) CWQNVSHSC(1) [WF]-G-[EQS]-N 6 Phage display (common panning) 3 PMID:9973494 Anti-lactotransferrin monoclonal antibody mAbRWL2 Lactotransferrin Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 146 SMSIASPYIPWS SPGPMKLLKTPL TLNINRLILPRT SMSIASPYIALE SMSIGSPYITFG TLNINRLILPRT 6 Phage display (subtractive panning) 3 PMID:16188121 Human gastric cancer cells GC9811-P Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Wild type cells GC9811 were carried out subtractive panning. After three rounds of screening, 40 phage clones bond to GC9811-P cells were randomly selected. When injected into the peritoneal cavity of nude mice, 6 of the 40 clones did not bind to mouse peritoneum as examined by immunohistochemical staining. They were considered to be capable of binding specifically to GC9811-P cells. 147 CYRGVTLAGHRC CYRGARVDGLMC CFKGVRLDGTPC CAREYGTNRWVC CFRGLDVAGNVC 5 Phage display (common panning) 4 PMID:16150491 Anti-parvalbumin polyclonal antibody Parvalbumin beta Not determined. Not determined. CL10 phage display library (CX10C) 148 SPSAAE SIYAAEK SYAAVAS SIQTKFAP QTVKFAP EGSGTTK SNRTQGE NLTQTQM YLENATQ TNPTQTK NPTQGSP NMTQAAQ HLYNPTQ SLPNLTQ TNPTQMA NPNQTRM TFGNRTQ NATQAST SNPTQGH SITNPTQ NSTQMRI SSPNTTQ AVTNATQ IPPNPSQ NPTQIRR NATQERY NPTQHSY ANITQSS SGPNPTQ NPTQRPD LNLTQSR FQINPTQ NPSQGPL INPTQTT THANSTQ SGPNATQ LAANLTQ LGPNHTQ NSTQGPV GSPNFTQ NPTQSPP QPPNITQ QGRNPTQ NQTQHSV YLPNVTQ HLPNATQ STPNVTQ NITQEAF TTLDANQ FSTQKSI LNFDPPS YALHSTP STPRLPD LPSYLQE GSSLHDI YTHNETE MTSSETQ GGQMRQG ANILSRR RVLTSMD FTPQMVR LLATQPA FNVTQGP 63 Phage display (common panning) 4 PMID:15543570 Anti-Nipah virus polyclonal antibody Nucleoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) The library was used to isolate peptides that interact with swine anti-Nipah virus antibodies immobilized on protein A-agarose. 149 CTALMSASC(6) CLREQPQQC(1) 2 Phage display (common panning) 4 PMID:15863836 Pancreatic triacylglycerol lipase Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In each round, bound clones were sequentially eluted with glycine-HCl, tetrahydrolipstatin, glycine-HCl and tetrahydrolipstatin. Four rounds of biopanning were performed. With the exception of the first round of each selection protocol, only the phages collected in the last elution step were amplified for the next round of biopanning. 150 CTALMSASC(2) 1 Phage display (common panning) 4 PMID:15863836 Pancreatic triacylglycerol lipase Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In each round, bound clones were sequentially eluted with glycine-HCl and tetrahydrolipstatin. Four rounds of biopanning were performed. With the exception of the first round of each selection protocol, only the phages collected in the last elution step were amplified for the next round of biopanning. 151 CTALMSASC(3) 1 Phage display (common panning) 4 PMID:15863836 Pancreatic triacylglycerol lipase Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In each round, bound clones were sequentially eluted with glycine-HCl, porcine pancreatic lipase and tetrahydrolipstatin. Four rounds of biopanning were performed. With the exception of the first round of each selection protocol, only the phages collected in the last elution step were amplified for the next round of biopanning. 152 CPPSYNGKC(3) CPDANRINC(1) CSQAPTPAC(1) CTPTSPAMC(1) CQPHPGQTC(1) 5 Phage display (common panning) 4 PMID:15863836 Pancreatic triacylglycerol lipase Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In each round, bound clones were eluted with tetrahydrolipstatin. Four rounds of biopanning were performed. With the exception of the first round of each selection protocol, only the phages collected in the last elution step were amplified for the next round of biopanning. 153 CSQLQTTKC(1) 1 Phage display (common panning) 4 PMID:15863836 Pancreatic triacylglycerol lipase Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In each round, bound clones were eluted with porcine pancreatic lipase. Four rounds of biopanning were performed. With the exception of the first round of each selection protocol, only the phages collected in the last elution step were amplified for the next round of biopanning. 154 SLFDTLA(10) SVFDRLA(3) LYAAINL(2) LPSFRAL(1) SPQLSKL(1) ALNLGSR(1) AINLGHR(1) AVRPVTP(1) PEKQAAR(1) GPPPKSH(1) TLFDTLK(1) TLFDMLR(1) TLFDVFR(1) SVFDTLA(1) AIWQKSW(1) SFHTFLQ(1) 16 Phage display (common panning) 3 PMID:16006240 Anti-EcA polyclonal antibody L-asparaginase 2 Not determined. Not determined. Ph.D.-7 phage display library (X7) 155 WSAPVLMGTVPP(8) QLNKLQIPLSII(2) RKVPLLSGTLPQ(1) QEPLMGTVPIRA(1) THNHQVPLAIIS(1) TMPWNQSALTLI(1) TLQQNMPLALVW(1) DPEQPAPLFLVS(1) APIQAHPLGLIR(1) PSIIGGSSVDLV(1) IDIINPAQNRLR(1) 11 Phage display (common panning) 3 PMID:15560754 Anti-NPS polyclonal antibody Neutral polysaccharides, NPS Not determined. Not determined. Ph.D.-12 phage display library (X12) 156 GPEDTSRAPENQQKTFHRRW(5) 1 Phage display (common panning) 5 PMID:15585132 Langerhans cells Not determined. Not determined. Not determined. X20 fAFF1 phage display library 157 ELRWDLTDIYNL(5) SWNSVDIWHMSS(4) W-x(3)-D-I 2 Phage display (common panning) 4 PMID:15483241 Anti-lycoprotein G monoclonal antibody MnAb1 Glycoprotein G Not determined. Not determined. Ph.D.-12 phage display library (X12) Conserved sequence, WxxxDI, aligned between aa 14 and 19 of the mature glycoprotein. 158 SWNSVDIWHMSS(4) SWSTLDIWSIPH(2) W-x(3)-D-I 2 Phage display (common panning) 4 PMID:15483241 Anti-lycoprotein G monoclonal antibody MnAb2 Glycoprotein G Not determined. Not determined. Ph.D.-12 phage display library (X12) Conserved sequence, WxxxDI, aligned between aa 14 and 19 of the mature glycoprotein. 159 ESLRWDQRIG(9) EFLGWGKAV(8) EVFRWSAASV(8) EEMRWGRLLS(5) NETRRWDILI(5) GGEEWFRWA(5) KDETWRWGR(4) LLRWGVSAT(4) EILRWGAIAP(3) VGRWGDFWL(3) PELWHTPV(3) VEMMRWGQTA(3) TEAFRWDMRA(2) LEHFRWMKPI(2) DYVCWWKASV(2) DGRWGDFWL(2) EWLRWLIYP(1) FCAVRWGLPC(1) LLRWGGLAV(1) SPRRSERLGW(1) ELFRWDTTHT(1) RIENFRWQRP(1) DSLRWGKVPV(1) ELAHPFWERY(1) SPTWRWAPY(1) WFRWGTAPAP(1) GYLRWHVPHS(1) LEQMRWMKHT(1) AELFRWGSSA(1) PQWFSWWRPA(1) 30 Phage display (common panning) 3 PMID:15893405 Anti-ACTH monoclonal antibody A2H8 Corticotropin Not determined. Not determined. X10 phage display library 160 SYGHPLGKPH(13) TMRWGFPVGR(5) SSGPPRAPGG(4) YTWVAVQSSR(3) HWGWPLGRLT(3) VWYQFPVGK(2) FGTPIGKPLR(2) FSAPVGKPGK(2) SYGRPLGKRL(2) GFGWPVGLKV(1) 10 Phage display (common panning) 3 PMID:15893405 Anti-ACTH monoclonal antibody 57 Corticotropin Not determined. Not determined. X10 phage display library 161 YGGEGLVSWG(2) YLENLSLPNR(2) SYGHPLGLPQ(1) WGSPVGLGH(1) FSAPVGLPGL(1) 5 Phage display (common panning) 3 PMID:15893405 Anti-ACTH monoclonal antibody FZ10-A32 Corticotropin Not determined. Not determined. X10 phage display library 162 WIATWQDDGYMY[0.704] 1 Phage display (common panning) PMID:15047171 Geminin Not determined. Not determined. Not determined. X12 phage display library ELISA The absorbance value (A) at 410 nm was determined by subtraction of the raw A of the GST plates from that of the GST–Geminin plates. Data shown were reproduced from the graph. 163 SFDPGLF(3) VEFPVSD(2) SIEFPTS(1) IEWPVSD(1) TIEFPVA(1) YAPEFPL(1) RHTMIIP(1) TIIFPVE(1) KWHWSDS(1) RHTMINP(1) NEFPVS(1) 11 Phage display (common panning) 3 PMID:15774614 Anti-GPIIIa49-66 polyclonal antibody Integrin beta-3 Not determined. Not determined. Ph.D.-7 phage display library (X7) 164 SHWETPLLSWFF SICDSHHRYSWL NHDYWYWLFENV WHSDMEWWYLLG WHVDETWLLMLT WHDPTPWWSWEI GNWMDFHLQRTA 7 Phage display (common panning) 4 PMID:15197767 Vascular endothelial growth factor receptor 1, VEGFR-1 Vascular endothelial growth factor A, VEGF-A 1FLT,1QTY, Not determined. Ph.D.-12 phage display library (X12) Peptide WHSDMEWWYLLG almost abolished VEGF binding to receptor Flt-1 in vitro. 165 TYPIPIR(3) TYPIPFR(3) TYPVPHR(1) TYPLPIR(11) TYPPPTR(1) T-Y-P-x-P-x-R 5 Phage display (common panning) 4 PMID:14532093 Bacillus anthracis Spore Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 166 NIMRNTW(4) AVPRASF(4) LGSYNNA(2) 3 Phage display (subtractive panning) 5 PMID:16599583 Lipopolysaccharide, LPS Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage library was first incubated in a well not coated with LPS for 1 h at room temperature, upon which the unbound fraction was transferred to LPS-coated wells. 167 ASFPPAF(4) SSHTISF(2) 2 Phage display (subtractive panning) 5 PMID:16599583 Lipopolysaccharide, LPS Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The panning procedure was the same as MS00166, except that an additional incubation step with LPS-O111 was added in each panning, and the unbound fraction was used for panning with LPS-O55. 168 RDFYYN(5)[+][NT] HDFFFN(2)[+][5.1 ± 0.3] WDLWVN(1)[+][3.4 ± 0.2] PGRYIN(1)[+][5.7 ± 1.1] NNYFFV(1)[+][0.006] GSGSIN(1)[+][0.9 ± 0.1] MPSYIN(1)[+][3.5 ± 0.2] PPAYFN(1)[+][1.8 ± 0.5] PDLYIN(1)[+][4.5 ± 0.1] MPQYVN(1)[+][2.0 ± 0.6] RGFYYN(1)[+][6.6 ± 1.6] NNYFFN(1)[+][6.6 ± 1.0] ARMYIN(1)[+][4.0 ± 0.2] DDTYNS(1)[+][1.8 ± 0.2] RGLYYN(1)[+][5.6 ± 0.1] 15 Phage display (common panning) 4 PMID:7499368 Cardiac glycoside digoxin Anti-digoxin antibody 26-10 Fab Not determined. Not determined. X6 phage display library ELISA,Saturation equilibrium assay Secreted Fab was tested using an ELISA with goat anti-mouse Fab, digoxin-BSA, and the congener-BSA used for selection as coating antigens. All clones were positive for Fab. Besides the designation + or - refers to whether the clone binds to the congener used in selection. Affinities (Ka,e-9 M-1) for digoxin of Fab 26 -10 and mutants (randomly mutated at positions H30 -35 inclusive) selected from the bacteriophage library. Affinities were measured using an equilibrium saturation method with filtration through glass fiber filters for separation of bound and free ligand, except that Fab was immobilized on the filter using goat anti-mouse Fab. 169 PSFYYN(1)[+][21.7 ± 8.1] RDFYYN(18)[+][9.7 ± 3.8] 2 Phage display (common panning) 4 PMID:7499368 16-acetylgitoxin Anti-digoxin antibody 26-10 Fab Not determined. Not determined. X6 phage display library ELISA,Saturation equilibrium assay Secreted Fab was tested using an ELISA with goat anti-mouse Fab, digoxin-BSA, and the congener-BSA used for selection as coating antigens. All clones were positive for Fab. Besides the designation + or - refers to whether the clone binds to the congener used in selection. Affinities (Ka,e-9 M-1) for digoxin of Fab 26 -10 and mutants (randomly mutated at positions H30 -35 inclusive) selected from the bacteriophage library. Affinities were measured using an equilibrium saturation method with filtration through glass fiber filters for separation of bound and free ligand, except that Fab was immobilized on the filter using goat anti-mouse Fab. 170 RDFYYN(4)[+] RRGIVN(2)[+] SKRYIN(1)[+] SMQWFN(1)[+] KDMYIN(1)[+] PFRFIN(1)[+] 6 Phage display (common panning) 5 PMID:7499368 Gitoxin Anti-digoxin antibody 26-10 Fab Not determined. Not determined. X6 phage display library ELISA Secreted Fab was tested using an ELISA with goat anti-mouse Fab, digoxin-BSA, and the congener-BSA used for selection as coating antigens. All clones were positive for Fab. Besides the designation + or - refers to whether the clone binds to the congener used in selection. 171 RDFYYN(7)[+][NT] GERFFN(2)[+][4.3 ± 0.6] SKRYIN(1)[+][0.3 ± 0.03] 3 Phage display (common panning) 6 PMID:7499368 Gitoxin Anti-digoxin antibody 26-10 Fab Not determined. Not determined. X6 phage display library ELISA,Saturation equilibrium assay Secreted Fab was tested using an ELISA with goat anti-mouse Fab, digoxin-BSA, and the congener-BSA used for selection as coating antigens. All clones were positive for Fab. Besides the designation + or - refers to whether the clone binds to the congener used in selection. Affinities (Ka,e-9 M-1) for digoxin of Fab 26 -10 and mutants (randomly mutated at positions H30 -35 inclusive) selected from the bacteriophage library. Affinities were measured using an equilibrium saturation method with filtration through glass fiber filters for separation of bound and free ligand, except that Fab was immobilized on the filter using goat anti-mouse Fab. NT denoted not tested. 172 SHSYIN(3)[+] RTRYIN(1)[+] TRYWFN(1)[+] PWTLLN(1)[+] SNLEDS(1)[-] SGRPPN(1)[-] YNRSQA(1)[-] LPPTSN(1)[-] 8 Phage display (common panning) 5 PMID:7499368 16-formylgitoxin Anti-digoxin antibody 26-10 Fab Not determined. Not determined. X6 phage display library ELISA The designation + or - refers to whether the clone binds to 16-formylgitoxin used in selection. 173 SHSYIN(6)[+][5.6 ± 1.7] TRYWFN(2)[+][3.9 ± 0.6] 2 Phage display (common panning) 6 PMID:7499368 16-formylgitoxin Anti-digoxin antibody 26-10 Fab Not determined. Not determined. X6 phage display library ELISA,Saturation equilibrium assay Secreted Fab was tested using an ELISA with goat anti-mouse Fab, digoxin-BSA, and the congener-BSA used for selection as coating antigens. All clones were positive for Fab. Besides the designation + or - refers to whether the clone binds to the congener used in selection. Affinities (Ka,e-9 M-1) for digoxin of Fab 26 -10 and mutants (randomly mutated at positions H30 -35 inclusive) selected from the bacteriophage library. Affinities were measured using an equilibrium saturation method with filtration through glass fiber filters for separation of bound and free ligand, except that Fab was immobilized on the filter using goat anti-mouse Fab. 174 IRKTR(3) TRKRR(2) PRPKR(1) IRRPR(1) PRTRR(1) PRPRR(1) YRLPR(1) KRLPR(1) KRTPR(1) RRTPR(1) GRQPR(1) GRIKR(1) PRLRR(1) KRTSR(1) SRHIR(1) VRVAR(1) KRTTR(1) TRIKR(1) RKKRD(1) RNRRS(1) RIKRW(1) KGNLE(1) TSPPL(1) TVRSR(1) KLENR(1) IGDDL(1) CTFTT(1) R-x(2)-R 27 Phage display (common panning) 6 PMID:7987214 Furin Not determined. Not determined. Not determined. AA-xxxxx-AA phage display library 175 LRRFRRP(5) LERVKRY(1) LVRRKRL(1) LPRGKRR(1) PRRFKRP(1) LVRSRRW(1) SRRFRRP(1) DRRFRRP(1) WFRPRRS(1) PRRPRRG(1) IGRPRRS(1) SRRYRRP(1) [LP]-R(2)-F-[KR]-R-P 13 Phage display (common panning) 5 PMID:7987214 Furin Not determined. Not determined. Not determined. AA-xxRx(K/R)Rx-AA phage display library 176 TQRIPRD(1) HIRKPRW(1) PRRRPRA(1) LGRSPRY(1) VRRGPRS(1) VSRYPRS(1) RHRSPRH(1) DDRSPRL(1) PVRAPRS(1) KSRRPRE(1) RRRPPRA(1) LRRIPRH(1) FSRVPRH(1) SARSPRC(1) KTRIPRG(1) SRRSPRS(1) 16 Phage display (common panning) 5 PMID:7987214 Furin Not determined. Not determined. Not determined. AA-xxRxPRx-AA phage display library 177 WHWRHRIPLQLAAGR(0.67) THSHQWRHHQFPAPT(0.24) HASHFRFRHSHVYGV(0.09) 3 Phage display (common panning) 3 PMID:12486137 Fibronectin type-II 1 domain of MMP-2 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Microtiter plates were coated with the recombinant proteins, consisting of the appropriate FN2 module and an amino-terminal peptide derived from the beta-galactosidase moiety of the expression vector. Bound phage were eluted with TBS buffer containing gelatin type A from porcine skin type I collagen (Sigma). 178 WHWRHRIPLQLAAGR(0.48) THSHQWRHHQFPAPT(0.46) HASHFRFRHSHVYGV(0.06) 3 Phage display (common panning) 3 PMID:12486137 Fibronectin type-II 2 domain of MMP-2 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Microtiter plates were coated with the recombinant proteins, consisting of the appropriate FN2 module and an amino-terminal peptide derived from the beta-galactosidase moiety of the expression vector. Bound phage were eluted with TBS buffer containing gelatin type A from porcine skin type I collagen (Sigma). 179 ACGYTYHPPCARLTV(0.03) WHWRHRIPLQLAAGR(0.25) THSHQWRHHQFPAPT(0.55) HASHFRFRHSHVYGV(0.17) 4 Phage display (common panning) 3 PMID:12486137 Fibronectin type-II 3 domain of MMP-2 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Microtiter plates were coated with the recombinant proteins, consisting of the appropriate FN2 module and an amino-terminal peptide derived from the beta-galactosidase moiety of the expression vector. Bound phage were eluted with TBS buffer containing gelatin type A from porcine skin type I collagen (Sigma). 180 ACGYTYHPPCARLTV(0.71) WFPGPITFIPRPWSS(0.16) THSHQWRHHQFPAPT(0.06) HASHFRFRHSHVYGV(0.06) 4 Phage display (common panning) 3 PMID:12486137 Fibronectin type-II 1-3 domain of MMP-2 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Microtiter plates were coated with the recombinant proteins, consisting of the appropriate FN2 modules and an amino-terminal peptide derived from the beta-galactosidase moiety of the expression vector. Bound phage were eluted with TBS buffer containing gelatin type A from porcine skin type I collagen (Sigma). 181 ACGYTYHPPCARLTV(0.25) WFPGPITFIPRPWSS(0.13) THSHQWRHHQFPAPT(0.37) WHVSPRHQRLFHGLF(0.25) 4 Phage display (common panning) 3 PMID:12486137 Fibronectin type-II 1-3 domain of MMP-2 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Microtiter plates were coated with the recombinant proteins, consisting of the appropriate FN2 modules and an amino-terminal peptide derived from the beta-galactosidase moiety of the expression vector. Bound phage were eluted with buffer containing recombinant protein described previously. 182 ACGYTYHPPCARLTV(0.88) THSHQWRHHQFPAPT(0.06) HASHFRFRHSHVYGV(0.06) 3 Phage display (common panning) 3 PMID:12486137 Fibronectin type-II 1-3 domain of MMP-2 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Phage were incubated with Protein-Sepharose. The recombinant protein consists of the appropriate FN2 modules and an amino-terminal peptide derived from the beta-galactosidase moiety of the expression vector. The bound phage were eluted with TBS buffer containing gelatin. 183 ERVREYYPEP SSMRKSFPEP GRIAPEPGGG WGNARKSPEP RVNPEPGRWA HITRLSPEPR RMRIDPEPRH VPRVRWSPEP PRFIEPRVSR WVSLRQVEPG PRFVEPRKET PRFIEPEGVS KYIEPGLRVR KYIEPRFLAM KYLEPWYACG LRCPEPGCLL CRTEPGCYVR CSSRVEPRAC ACRSEPRCFP FCKAEPLCAR R-x(3)-P-E-P, R-x(2)-P-E-P, R-x(2)-E-P, K-x(2)-E-P , C-x(n)-C 20 Phage display (common panning) 2 PMID:15568679 Anti-colicin-A monoalonal antibody 1C11 Colicin-A Not determined. Not determined. X10 phage display library 184 CWEDGWLQTC CWEDAWLGLC CTLWMEDGWC CTEDGWLMTC CILWMDDGWC CWDDGIWFC CWDDAWWWC CPEELWWLC CWQEWDGWEC CWQDWDGWDC CFMWHDWLVC CWVDSWWGC ADWYMFVYGS [DE]-[DE]-[GL]-W 13 Phage display (common panning) 3 PMID:12824186 Integrin alpha-M, integrin Not determined. Not determined. Not determined. CX(7-10)C and X(9-10) phage display library pool Such negatively charged sequences are present in many known β2 integrin ligands and also in the catalytic domain of matrix metalloproteinases (MMPs). 185 CSGLRNETFLRC CEFFQQHMLRVPRC CNMKLKLREMTQRC CMTRPTSLTQLTGC CLHIRVNETAYRVC CDFLREHGMKNPRC CRSRPTNMTTLRDC CAAYNATRGTVSAC CQLLHTWEDKMRKC CRNGELWLRRPGLC CMVRPSNWDALTRC 11 Phage display (common panning) PMID:14596802 Anti-gp120 monoclonal antibody 17b Surface protein gp120 (SU) 1GC1,1G9M,1G9N,1RZJ,1RZK,2NXY,2NXZ,2NY0,2NY1,2NY2,2NY3,2NY4,2NY5,2NY6, Not determined. Three CX12C phage display library pool 186 CAHFPPRSQMIADC CAHFAPGTAMYSDC CRQFPHSSSMYTDC CRESRAALERGWWC CEARTHNEARRRRC CAAARSTGETSAHY CYYRMGANYTVGEC CSVSPLYAYDDPLC CTQMHEMDPNFPPC CVTALGPNYTGQEC CYVQQPWWVLEREC CADVMGPLVTAAEC CADVMGPLVTAGEC CVVFLDVSEAFRDC VWRCNWF AASWNGR 16 Phage display (common panning) PMID:14596802 Anti-P24 monoclonal antibody 13b5 Capsid protein p24 1E6J, Not determined. CX12C phage display library 187 CAKEGDLNKYKPWC CMRLPIDNRYRPWC CRYAVTVPQKNRYC CQFDPEIDWIPPKC CRSDTTVGWRPPRC CVWETGSRWRMPKC CKGVHVEDQYRPWC CQWETNGGWTRPKC CWSPDSAWEKPKCC CWEGPVSPAGLPRC CEYDNQSHWARPKC CGMTLRGWRDPRMC CRVAQAEGWAPPRC CIRNYHVFPPGKVC CAKNYLLYPPIRQC CRTGTVSWGQYKGC CKLDYRIGENFAQC CKYEVSQEARERWC CGTHPYGSPQFMTC CLVGETRLGIWLGC CSSSCAKWAWSHCC CRGGGTDAQGWYTC CRRPVVDAGWNMSC CSGASAWRGEMQHC CKDLAEGGPAIGSC CYKPLYAQGWGWYC CSPRTRTGTARSRC CDPKRNPDRIMHPC 28 Phage display (common panning) PMID:14596802 Anti-gp120 monoclonal antibody CG10 Surface protein gp120 (SU) Not determined. Not determined. CX12C phage display library 188 CLFNLPWLC(12)[0.263 ± 0.149] CLFDLPWLC(3)[0.123 ± 0.003] CMFNLPWLC(2)[0.299] CPFKHLPWC(2)[0.381 ± 0.113] CSFTWLPWC(1)[0.173 ± 0.181] CPFQYLPWC(1)[0.247 ± 0.101] CPFQFLPWC(1)[0.411 ± 0.080] CPFSFLPWC(1)[0.410 ± 0.128] C-x-F-x-L-P-W-L-C, C-x-F-x(2)-L-P-W-C 8 Phage display (common panning) 3 PMID:14978012 T-cell surface glycoprotein YE1/48 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance at 492 nm from a triplicate study were determined. Data were reproduced from the graph and shown as means ± SD. 189 CGAVIDDC(5) CGVVVLQC(1) CGELGVDC(1) 3 Phage display (common panning) 2 PMID:15049781 Anti-LPS monoclonal monoclonal MoAb 9-2-L3,7,9 L3,7,9 lipopolysaccharides (LPS) Not determined. Not determined. Cys6 phage display library (CX6C) 190 CRGDDFPAWYC(3) CKLSQGAPDSC(3) CAQQWPEWYPC(1) CKPIDAERGLC(1) CADSWGLDLRC(1) P-x-W-Y 5 Phage display (common panning) 2 PMID:15049781 Anti-LPS monoclonal monoclonal MoAb 9-2-L3,7,9 L3,7,9 lipopolysaccharides (LPS) Not determined. Not determined. Cys9 phage display library (CX9C) 191 CPWWEDMC(4) CWWWEDEC(2) CRWWEDKC(1) CAWWEDVC(1) CDGFGWDC(1) CMWWEDGC(1) CLWWEDMC(1) CNFWWEDC(1) CRWWDDFC(1) CYWWSDRC(1) CYWWSDWC(1) CWWWSDYC(1) CWWWSERC(1) CWWWMDDC(1) CWWWVDRC(1) CYWWLDSC(1) CAWWADVC(1) W(2)-E-D 17 Phage display (common panning) 2 PMID:15049781 Anti-LPS monoclonal monoclonal MoAb 1-6-C L3,7,9 lipopolysaccharides (LPS) Not determined. Not determined. Cys6 phage display library (CX6C) 192 CPWWADNYWEC(6) CPWWEDNWYSC(1) CSAPLRWWDDC(1) CAVMWWDDGWC(1) CCTAWWCDQWC(1) P(0,1)-W(2)-x-D-N(0,1) 5 Phage display (common panning) 2 PMID:15049781 Anti-LPS monoclonal monoclonal MoAb 1-6-C L3,7,9 lipopolysaccharides (LPS) Not determined. Not determined. Cys9 phage display library (CX9C) 193 CDIFGRDC(3) CMGRTMTC(3) CDVWGRDC(2) CDVWGLDC(1) CLWGLDSC(1) CYWGLDNC(1) CWWALDGC(1) D-x-[FW]-G-x-D 7 Phage display (common panning) 2 PMID:15049781 Anti-LPS monoclonal monoclonal MoAb 4-2-C L3,7,9 lipopolysaccharides (LPS) Not determined. Not determined. Cys6 phage display library (CX6C) 194 CMGDAFGRDGC(3) CDLWGWDLAYC(1) CWFDRWGRDYC(1) CEDEWGGSLVC(1) CLEPLFGLDTC(1) CMEARLWGLDC(1) CYDPWGLVYAC(1) D-x-[FW]-G-x-D 7 Phage display (common panning) 2 PMID:15049781 Anti-LPS monoclonal monoclonal MoAb 4-2-C L3,7,9 lipopolysaccharides (LPS) Not determined. Not determined. Cys9 phage display library (CX9C) 195 CWGRTMTC(7) CWGRTMRC(1) CRGGPHTC(1) G-R-T-M-T(0,1) 3 Phage display (common panning) 2 PMID:15049781 Anti-LPS monoclonal monoclonal MoAbR3 L3,7,9 lipopolysaccharides (LPS) Not determined. Not determined. Cys6 phage display library (CX6C) 196 CGTSMTTRHLC(3) CGQTMRWSRGC(2) CGQSFRWRLRC(2) CGVTMTRGRLC(2) CGASMTLGRLC(2) CGAVYSMRVMC(2) CGQTMQLGLNC(1) CGQSMRGGRFC(1) CGQSMLWRAFC(1) CGQSFTMVRFC(1) CGSTMLMRRQC(1) CGASMQWGRWC(1) CGTSMTYMMRC(1) CGRTMSNRVAC(1) CGRSMVSGRWC(1) CGITLARGREC(1) CGDSFIFRNAC(1) CGWRWDLRASC(1) CGRVWSQRGYC(1) G-[xQ]-[ST]-M 19 Phage display (common panning) 2 PMID:15049781 Anti-LPS monoclonal monoclonal MoAbR3 L3,7,9 lipopolysaccharides (LPS) Not determined. Not determined. Cys9 phage display library (CX9C) 197 CGYGRFSPPC(6) CRWYGPILWC(2) CGWGRYSPPC(1) CGFGRWQPPC(1) CRVYGPYLLC(1) CRWYGPWALC(1) CRWYGPWVWC(1) CRFYGAWLLC(1) CRHYGPFSIC(1) CRRYGPFMVC(1) CRTYGWWVVC(1) CRYYGWLTVC(1) CKWYGLFQLC(1) CHSYGPFVVC(1) CNWYGWFRVC(1) C-G-[FYW]-G-R-[FYW]-[SQ]-P(2)-C, C-R-x-Y-G-P-x(3)-C 15 Phage display (common panning) 3 PMID:15123665 Matrix metalloproteinase-9, MMP-9 Integrin beta-5 Not determined. Not determined. CX(7-10)C and X(9-10) phage display library pool 198 WPRLLFDASANH(2)[0.301/0.376] TREFMKTASRCP(2)[0.407/0.415] NVQFLEIARRYS(2)[0.493/0.285] 3 Phage display (common panning) 3 PMID:15202505 Anti-ferritin polyclonal antibody Ferritin Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured at 490 nm. Data shown were reproduced from the graph. 199 AKIHFYS DKIHFKS VNKPHFH AYIHFLP SEIHLMG AKIHFHS HKPHFPS ALQIHTM AQIHFLP DNPHYTT GKPHFTT SFHIYFL SKIHFHT RKPHFTG LKLHFNT RKIHFHP x-K-I-H-F-x-S 16 Phage display (common panning) 4 PMID:18642701 Anti-FASLG receptor monoclonal antibody SA-8 Tumor necrosis factor receptor superfamily member 6 Not determined. Not determined. Ph.D.-7 phage display library (X7) 200 GKIHFFR(4) GYKPTFS(2) SKIHFHP(2) NKIHFYP(1) SKIHFLS(1) GKIHFHP(1) SKIHFPE(1) CKIHFFR(1) DKIHFYP(1) RYYPYFP(1) AKIHFPP(1) EKPHFLV(1) AKIHFLP(1) HKIHFLP(1) RKLHFLP(1) SKIHFYH(1) C-K-x(2)-F(2)-x 16 Phage display (common panning) 4 PMID:18642701 Anti-FASLG receptor monoclonal antibody SA-7 Tumor necrosis factor receptor superfamily member 6 Not determined. Not determined. Ph.D.-7 phage display library (X7) 201 YPRYDYHDW WSHYSSEPL PSMSYYTTW YQYIETSMQ CDWELCYAE YYYSTPSVA NYYDRYAIE VADLTYYAW AHNYYPREW 9 Phage display (common panning) PMID:15265865 Synapsin-1 Ras-related protein Rab-3A Not determined. Not determined. pVIII-9aa phage display library (X9) Antibodies raised against the peptide YQYIETSMQ (syn21) specifically recognized Rab3A, a synaptic vesicle-specific small G protein implicated in multiple steps of exocytosis. 202 EDPSHSLGLDVALFM(9)[0.428 ± 0.043][5.0 ± 0.4] QTPETLTLFPPNWMN(7)[0.399 ± 0.017][15.4 ± 2.2] EANAPLDMGVFFEHN(3)[0.371 ± 0.074][16.2 ± 1.5] NNNQLDCLLFVRQCK(1)[0.150 ± 0.060][112.0 ± 21.0] RLTSPADMGSFLFNL(1)[0.250 ± 0.051][227.0 ± 18.0] RTTLGHHLDFTLWTA(1)[0.147 ± 0.024][60.0 ± 5.4] 6 Phage display (common panning) 4 PMID:15833876 Anti-GD2 monoclonal antibody 14G2a GD2 ganglioside Not determined. Not determined. X15 phage display library ELISA,Flow Cytometry For reverse ELISA, 14G2a mAb-coated wells were incubated with positive phage clones (0.5e12 transforming units), washed and incubated with HRP-conjugated anti-M13 mAb. After a final wash, the reaction was developed with the HRP substrate and quantified by reading the absorbance at 450 nm in a plate reader. The wells incubated with negative clones remained nearly colorless. The absorbance was shown. Besides, synthetic peptides representing the individual phage families were analyzed for the ability to inhibit the binding of 14G2a mAb to GD2-positive IMR-32 neuroblastoma cell. Varying amounts of the competitors were incubated overnight with 14G2a antibody (1 μg/mL). The mixtures were transferred to IMR-32 cells, and the inhibition of 14G2a binding to cellular GD2 was measured after staining with FITC-conjugated secondary antibody by flow cytometry analysis. The IC50 (μmol/L) was shown. 203 FRMDFDYLYPSLP LNFMIFYLSLNPW FSYSVSYAHPEGL SVAFYDYLPTDLP LSFSDFYFSEGSE FAPMKSYGVSLPP LFGPIEYTQFLAN LFDAYWYSDTAMS PASLELYENLVAG GENFCPYSFFGCG YLSLHAYESFGGS FFGFDVYDMSNAL FYMPFGPTWWQHV LPHLIQYRVLLVS GFAWSSYLGTTVH FLSFVFPASAWGG FFPSSWYSHLGVL FFSFFFPASAWGS 18 Phage display (common panning) PMID:16873017 96-well immunoassay plate (polystyrene, PS) Not determined. Not determined. Not determined. X6PX6 and X6YX6 phage display library pool 204 FAQDLQG(16) WNTKASY(3) SEHSVRL(3) SAHPLYT(3) FNQDIMG(3) AHPQLAT(3) EVTSTAF(2) YTFTQDN(2) LNSKGPI(2) RPLRNWP(2) FSQDLHP(2) FNQDLHS(2) TNAPRMA(1) TSLQHLM(1) SSTPALL(1) MGHKYSQ(1) SPTSGHT(1) YNAPPQL(1) LPRPFTL(1) AVVPKSL(1) SLLNSPV(1) EPWYYSP(1) TFVRINP(1) AWYSLTW(1) MPFGDHH(1) ITNGPPM(1) SLAMYQY(1) DSRNSAT(1) LTLPAAL(1) TSLQHLV(1) SHTPALL(1) IPLQSLS(1) LHAGMPP(1) LAQRFST(1) TIANLSL(1) AEPVAML(1) SGFVFQQ(1) SISNLLL(1) YTFRQDM(1) 39 Phage display (common panning) 3 PMID:16054642 Anti-M. hyopneumoniae polyclonal antibody Mycoplasma hyopneumoniae Not determined. Not determined. Ph.D.-7 phage display library (X7) 205 CQTALNRFC(9) CSPNQLKLC(7) CLPSLSSGC(6) CKNLGVKEC(6) CSVRTDTHC(5) CPKPEQGRC(5) CNPFDHASC(4) CPVQNPRYC(4) CSPGDSPEC(4) CPQFLTNIC(3) CLHPWHKNC(3) CPQSLVRIC(2) CKQFLNMRC(2) CTLPSLRHC(2) CLFDRNNSC(2) CPQSLRHIC(1) CPEKMLAWC(1) CNPFNHSAC(1) CLQRLQQHC(1) CPKPEQLLC(1) CPQFLSNIC(1) CNSWLNMTC(1) CPQQLTRIC(1) CPQSLSRLC(1) CPQMLSRIC(1) CEGPAPEEC(1) CWLNMSKDC(1) CPQHLHRVC(1) CNPFNHQLC(1) CTLDRLKQC(1) CNPFNHSGC(1) 31 Phage display (common panning) 3 PMID:16054642 Anti-M. hyopneumoniae polyclonal antibody Mycoplasma hyopneumoniae Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 206 CPTSLLWWC CQRSPHSTC 2 Phage display (common panning) 5 PMID:15598577 Galectin-1, Gal-1 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 207 CQSPSARSC(6) CGAHRLHQC(3) CYSFPHTTC(1) 3 Phage display (common panning) 5 PMID:15598577 16 kDa beta-galactoside-binding lectin Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 208 FDLSEALVPPHR(7) ELDLADALPVLA(5) LPFNLTDALPWW(1) WHVTPFTLYDAM(1) SPGNFSLADALL(1) VVPFSLRDALAI(1) SSSYPLLLEALP(1) NTLGTFSLYDALG(1) PKDFSLFEALLG(1) CSAYALDALLPR(1) [AL]-x-[DE]-A-[LM] 10 Phage display (common panning) 4 PMID:15979564 Calpain-1 catalytic subunit Calpastatin Not determined. Not determined. Ph.D.-12 phage display library (X12) 209 ANDLAHWEPMSF(1) MCRLCHWPSASR(1) HSPTFHGLPPGL(1) AHLRPAPGDPWF(1) KLLPNSLWARTP(1) KLLPPPTLGRST(1) KMMPELNPSVVH(1) QIVKQNPNAVLS(1) QLPHRIPLIPDQ(1) 9 Phage display (common panning) 1 PMID:15927323 Anti-P25-LHRH polyclonal antibody Luteinizing hormone-releasing hormone I Not determined. Not determined. Ph.D.-12 phage display library (X12) 210 MYTQQRHWSEGL(1) TKIQEELAHWSP(1) DSSSMLHWTYSL(1) HWEYRVRTYPQT(1) APSLLHWGPSFW(1) GKLVWSDSFSTA(1) KLIPNSSLHSDS(1) SVPPKTIPNHSL(1) KLTPSQSLLGAT(1) TSKLTPGHSILN(1) SKLLPNYALTLN(1) STLKMLPHSRFF(1) MKVTPNRSLSTT(1) SKLMPNNSLWLY(1) KAIPGSWWQLWS(1) AKLPANWHLMLP(1) 16 Phage display (common panning) 2 PMID:15927323 Anti-P25-LHRH polyclonal antibody Luteinizing hormone-releasing hormone I Not determined. Not determined. Ph.D.-12 phage display library (X12) 211 KGIPNSMLLELR(2) KMSPTINSLLWQ(2) QYEHWSEGLT(1) GELDHWSGLPPH(1) DAPHMRHWSWAL(1) RPPLSELSHWHP(1) KLIPNSSLLMGA(1) HKMIPNHVLWRP(1) KILPNQSLFHVG(1) GKWLPNSSFMQH(1) INAKMLPNHFLT(1) HKLTPNWSLSFT(1) KITPNYSLVSGP(1) KLSPNAMLLEQP(1) KLSPNMTLLGPP(1) YAKQVPNTSLVM(1) NAPKLVSNWFLA(1) 17 Phage display (common panning) 3 PMID:15927323 Anti-P25-LHRH polyclonal antibody Luteinizing hormone-releasing hormone I Not determined. Not determined. Ph.D.-12 phage display library (X12) 212 TNTHWSYMLPSA(1) NVVSVGHWSYYV(1) TLHEHWSRALLE(1) HWSSMVHSPMP(1) SFGTHWSDLLSH(1) ATTMHWSDQLEA(1) HWSTRVHHTPNS(1) QTITTMHWSSSL(1) HWSDRVHNRLTL(1) HWNYYKDTLPIQ(1) KSLPLESFLAPW(1) FNDKHIPNSRLL(1) STFPKLPPGASL(1) QILVETQVKQTP(1) TQETTISGRVLP(1) 15 Phage display (common panning) 2 PMID:15927323 Anti-P25-LHRH polyclonal antibody Luteinizing hormone-releasing hormone I Not determined. Not determined. Ph.D.-12 phage display library (X12) Two rounds of panning experiments were conducted: the first with acid elution and the second eluted by LHRH peptide. 213 ASYSDLLYSSFL(15) ASYSDVLYSSFL(2) SDKLLHSVRPPV(2) 3 Phage display (common panning) 2 PMID:15927323 Anti-P27-LHRH polyclonal antibody Luteinizing hormone-releasing hormone I Not determined. Not determined. Ph.D.-12 phage display library (X12) The panning experiments were conducted using the generic acid elution procedure. 214 KLLNSVPLIHGS(8) NLLKSSNTLPFD(1) DSLHWSYMLLPS(1) 3 Phage display (common panning) 3 PMID:15927323 Anti-P27-LHRH polyclonal antibody Luteinizing hormone-releasing hormone I Not determined. Not determined. Ph.D.-12 phage display library (X12) The panning experiments were conducted using the generic acid elution procedure. 215 KLLNSDALPVVA(13) KQLNSVPQELSF(2) GVKQLNSVIKPG(1) 3 Phage display (common panning) 3 PMID:15927323 Anti-P27-LHRH polyclonal antibody Luteinizing hormone-releasing hormone I Not determined. Not determined. Ph.D.-12 phage display library (X12) A total of three rounds of panning experiments were conducted: the first two with acid elution and the third eluted by incubation in TBS buffer. 216 WPAALFESNRPD(18) DIHWTSMMVRPT(1) 2 Phage display (common panning) 2 PMID:15927323 Anti-P35-LHRH polyclonal antibody Luteinizing hormone-releasing hormone I Not determined. Not determined. Ph.D.-12 phage display library (X12) The panning experiments were conducted using the generic acid elution procedure. 217 CDRASPYC(4) CDLASPWC(4) CDRATPYC(3) CDRASPWC(3) D-[RL]-A-[ST]-P-[YW] 4 Phage display (common panning) 3 PMID:16923020 NeutrAvidin Biotin Not determined. Not determined. CX6C phage display library 218 CDRASPYC(12) CDRATPYC(7) CDLASPWC(3) CDRASPWC(2) D-[RL]-A-[ST]-P-[YW] 4 Phage display (common panning) 4 PMID:16923020 NeutrAvidin Biotin Not determined. Not determined. CX6C phage display library 219 CDRASPYC(20) CDLASPWC(7) CDRATPYC(6) CDRASPWC(2) D-[RL]-A-[ST]-P-[YW] 4 Phage display (common panning) 5 PMID:16923020 NeutrAvidin Biotin Not determined. Not determined. CX6C phage display library 220 CSSKSYRPC(5) CNKPKNASC(1) CPHLPNSTC(1) 3 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody P1 (2C3D4) Leptospira Not determined. Not determined. CX7C T7 phage display library 221 CTPKKSGRC(2) LTPCDSNKSGRC(2) TPCSKKSTRNC(1) TPCSKKDPRNC(1) CRKKNTNNC(1) CRKSKSASC(1) CTTNSKRKC(1) 7 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody P2 (8C6C4A12) Leptospira Not determined. Not determined. CX7C T7 phage display library 222 LTPCDPNTNSFC(5) CGIPGTPDC(3) LTPCDPNTNNFC(1) LTPCDINTNSFC(1) 4 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody P4 (2D2A12G3) Leptospira Not determined. Not determined. CX7C T7 phage display library 223 CKSKKSSSC(5) CNKKDPSSC(1) TPCSKKNPGNC(1) CKSKLVRC(1) CESNKSSC(1) 5 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody P5 (15BC2G1) Leptospira Not determined. Not determined. CX7C T7 phage display library 224 LTPCDAPPSGIC(9) CGRTRVTRC(1) 2 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody P6 (1B4D3F10) Leptospira Not determined. Not determined. CX7C T7 phage display library 225 CFATVERAC(7) CLLTVERAC(1) LTPCDSDWPVSC(1) 3 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody P7 (16B1C9H7) Leptospira Not determined. Not determined. CX7C T7 phage display library 226 CPPPKKGNC(1) LTPCDT(1) CKSKKSSAC(1) CERILWVVC(1) CNIRNAYTC(1) LTPCDRNTMVIC(1) LTPCDPNTSNFC(1) CCTNILVVC(1) 8 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody P9 (4A2D5D6) Leptospira Not determined. Not determined. CX7C T7 phage display library 227 CTKKKSSSC(1) CRKKISTIC(1) CRKKNPSNC(1) LTPCDKKYYGCC(1) LTPCDTNTYYCC(1) CRPNKKNAC(1) LTPCDT(1) TPCSPKKKGSC(1) CRKS(1) CCDSKYYLRC(1) 10 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody P10 (1A4B10E1) Leptospira Not determined. Not determined. CX7C T7 phage display library 228 TPCSTIYNDDC(3) CNNIYNDDC(3) CRTITNDKC(1) CPSIYNDDC(1) CTKKGPRNC(1) LTPCDTRDLVVC(1) 6 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody P17 (8C1E11) Leptospira Not determined. Not determined. CX7C T7 phage display library 229 CFNSTNDPC(2) CFNATNDPC(2) CPKSKSSRC(1) CTPKKNRAC(1) TPCSKKRSISC(1) CLTPLNDPC(1) 6 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody F11 (9B4C1D8) Leptospira Not determined. Not determined. CX7C T7 phage display library 230 TPCSTLINIFC(3) CFK(2) CRTKKTGSC(1) 3 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody F20 (9A4G1A8) Leptospira Not determined. Not determined. CX7C T7 phage display library 231 TPCSPKRKANC(1) 1 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody F21 (9A5A11A11) Leptospira Not determined. Not determined. CX7C T7 phage display library 232 LTPCDNY(5) 1 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody LD5 Leptospira Not determined. Not determined. CX7C T7 phage display library 233 CVLKKNRPC(3) CLP(2) 2 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody LF9 Leptospira Not determined. Not determined. CX7C T7 phage display library 234 LTPCDAERSGYC(1) LTPCDAPTFGSC(1) LTPCDPHFLDDC(1) LTPCDLRTSGYC(1) CRSKRIRNC(1) TPCILLSIGFC(1) CTTKRTPRC(1) LTPCDLRPFWFC(1) CRNSKSPNC(1) 9 Phage display (common panning) 4 PMID:16581206 Anti-leptospira monoclonal antibody P20 (7A5F12) Leptospira Not determined. Not determined. CX7C T7 phage display library 235 CVLSRSRSC(1) CRIK(1) CRTK(1) LTPCDLRRLGYC(1) TPCSSKRSTSC(1) TPCSPKASRSC(1) TPCI(1) LTPCDLRSLGYC(1) 8 Phage display (common panning) 4 PMID:16581206 Anti-leptospira polyclonal antibody 1 Leptospira Not determined. Not determined. CX7C T7 phage display library 236 TPCSKK(1) CRNPQHTNC(1) CVTNNTPDC(1) LTPCDLRRSGYC(1) CTTKKNVTC(1) CFNPLNDNC(1) CAPNSNRRC(1) LTPCDTKKFGYC(1) 8 Phage display (common panning) 4 PMID:16581206 Anti-leptospira polyclonal antibody 2 Leptospira Not determined. Not determined. CX7C T7 phage display library 237 TPCSTKRSASC(2) CVPKKKGNC(1) CPYPIITFC(1) CKTKRSASC(1) CGSSRLAIC(1) LTPCDNN(1) CLNPLKVYC(1) CASSSIVDC(1) 8 Phage display (common panning) 4 PMID:16581206 Anti-leptospira polyclonal antibody 3 Leptospira Not determined. Not determined. CX7C T7 phage display library 238 TPCILSGSASC(1) TPCSNKSKRNC(1) TPCSLTVTTIC(1) CRKKNLGNC(1) PCNTKRSASC(1) CTL(1) CKTKRSASC(1) CRSKNPANC(1) PCNTKRTASC(1) 9 Phage display (common panning) 4 PMID:16581206 Anti-leptospira polyclonal antibody 4 Leptospira Not determined. Not determined. CX7C T7 phage display library 239 WEADDKNQHGEG ISLTEWSMWYRH EEGPWSTHVGRT WGNEGGDHLQPV SLKIRWELKMYQE AVERWEKHTWSE 6 Phage display (common panning) PMID:17446277 Anti-ManLAM monoclonal antibody CS40 ManLAM Not determined. Not determined. X12 phage display library 240 GGSGTSRTPILG(7) AVGLSPDGSRGV(2) TLPIISTSLMRH(2) GHFSYPYARQTP(1) QLCIFHNEWNPV(1) SHLVYLTRHCVD(1) 6 Phage display (common panning) 3 PMID:16359760 Anti-RAPs polyclonal antibody 50S ribosomal protein L2 Not determined. Not determined. Ph.D.-12 phage display library (X12) 241 FVDILFAGMSGA(7) MHNPSMT(4) MHSPLPI(2) FVEIYSP(1) FVEHTRW(1) FVLPWRI(1) MHTPAIG(1) MHLNTPP(1) FIDWQSR(1) FIEVPWK(1) MHSPAAT(1) MHTPNSR(1) MHLNTAS(1) MHSPSNA(1) MHLTQSP(1) MHLTTNI(1) FVLVLFAGMSGA(1) 17 Phage display (common panning) 3 PMID:16360019 Anti-PDC-E2 polyclonal antibody Dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex, mitochondrial Not determined. Not determined. Ph.D.-12, Ph.D.-7 and Ph.D.-C7C phage display library pool Two independent biopannings were performed using the PhD-7 random heptapeptide library (New England Biolabs, Beverly, MA) and IgG from two PBC sera, and three independent biopannings using a mixture of libraries comprising the PhD-7 library, a cysteineconstrained heptapeptide library PhD-C7C, and a 12mer library PhD-12 (New England Biolabs, MA, USA), and IgG from a further three PBC sera. Thus, in all, phage clones were available from five separate biopannings with sera from five patients. 242 QDKLTQWPKWLE(8) NMSIQWHRDFNK(4) HEENDLRWWFIH(2) DDNTKQWPSWLE(2) DRAQLTWPAWLE(2) SPYQYSWPTWLE(1) STNMGAWPWWLE(1) 7 Phage display (common panning) 3 PMID:16814270 Anti-CD20 monoclonal antibody rituximab B-lymphocyte antigen CD20 2OSL, Not determined. Ph.D.-12 phage display library (X12) 243 WHWTWLSEYPPP(3) GGPIPTIPRPTL(2) WHWTWLSDYPPP(1) WQWTWLCEHPPP(1) WHRPPWWPSSSL(1) HELRSQIISTTS(1) FQWSWYTPPRPS(1) 7 Phage display (common panning) 3 PMID:16713681 Anti-Gb3 polyclonal antibody Globotriaosylceramide (Gb3Cer) Not determined. Not determined. Ph.D.-12 phage display library (X12) 244 WHWTWLSEYPPP(3) EQWPWSLLPIHY(2) NALTSHTASPAP(1) HPPGHRLTLIPH(1) VPSLHQTTHPTP(1) HPSCNSATGIPH(1) 6 Phage display (common panning) 4 PMID:16713681 Anti-Gb3 polyclonal antibody Globotriaosylceramide (Gb3Cer) Not determined. Not determined. Ph.D.-12 phage display library (X12) 245 KHMQNPF(7) GHMSNPF(3) SHLANPF(2) SHLSNPF(2) DHLRNPF(1) FHQPNPF(1) LHLRNPF(1) KHLINPF(1) QHLQNPF(1) SHNPNPF(1) THLTNPF(1) QHLLNPF(1) WHNSNPF(1) KHLSDPF(1) H-x(2)-[ND]-P-F 14 Phage display (common panning) 3 PMID:16904109 Recombinant protein N36(L8)C34 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 246 NHLHNPFTVDSG(1) SHLHNPFAFPAQ(1) DHTTNPFGTPLP(1) SHTENPFTVPQQ(1) FHLDNPFLADRS(1) AHTTDPFQPWAL(1) SHLSDPFGELRP(1) GHNQFNPFTFVM(1) GHNHVLNPFKVY(1) H-x(2)-[ND]-P-F 9 Phage display (common panning) 3 PMID:16904109 Recombinant protein N36(L8)C34 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 247 SNWKWWPGIFD HWWRSWYSDSV HEWHWWHQEAA WGLEHFAGNKR WWRWNWATPVD WHNYFHWWQDT 6 Phage display (common panning) 4 PMID:17046401 Polyglutamine containing 62 consecutive glutamines Not determined. Not determined. Not determined. X5-Fixed-X5 phage display library 248 CIPSHTPRC(4) CHPQYHARC(1) CGPMGISTC(1) CSGPLQRSC(1) CYSAPVSAC(1) CQATPHPXC(1) CNKERPWVC(1) 7 Phage display (common panning) 3 PMID:16517013 UDP-N-acetylmuramoylalanine--D-glutamate ligase ATP, D-glutamate Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Phage encoding peptides were eluted in the each round of biopanning using glycine-HCl. 249 HLPTSSLFDTTH(2) NPHWSSLYAPRN(2) TSYIPSFPATKY(1) ERPQPLDPQTFN(1) DAKNSNSRILDQ(1) DHQRMNDAMLVL(1) HVTTTFAPPPPR(1) 7 Phage display (common panning) 3 PMID:16517013 UDP-N-acetylmuramoylalanine--D-glutamate ligase ATP, D-glutamate Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage encoding peptides were eluted in the each round of biopanning using glycine-HCl. 250 CIPSHTPRC(1) CHPQYHARC(1) CTPHLPLRC(1) CSPTLRTIC(1) CNTSSSPRC(1) CPHKNNSTC(1) CPMLDNHKC(1) CNQRLTGEC(1) CSFRLSLHC(1) CYSAPVSAC(1) 10 Phage display (competitive panning) 3 PMID:16517013 UDP-N-acetylmuramoylalanine--D-glutamate ligase ATP Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Phage encoding peptides were eluted in the each round of biopanning using glycine-HCl, glycine-HCl and ATP. 251 HLPTSSLFDTTH(1) NPHWSSLYAPRN(1) HTQNMRMYEPWF(1) GQSPHSYQPRTY(1) TPSVLSTALHSS(1) SLTHAWQQTHFL(1) WSVTNLVLLSPP(1) FAKNSNSRILDQ(1) YQLRPNAESLRF(1) SNWYNGLEFLET(1) 10 Phage display (competitive panning) 3 PMID:16517013 UDP-N-acetylmuramoylalanine--D-glutamate ligase ATP Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage encoding peptides were eluted in the each round of biopanning using glycine-HCl, glycine-HCl and ATP. 252 CIPSHTPRC(5) CGPMGISTC(1) CSSMPLPAC(1) CSSSPMRTC(1) CRDTLFSQC(1) CLRSAGPSC(1) 6 Phage display (competitive panning) 3 PMID:16517013 UDP-N-acetylmuramoylalanine--D-glutamate ligase D-glutamate Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Phage encoding peptides were eluted in the each round of biopanning using glycine-HCl, glycine-HCl and D-Glu. 253 HLPTSSLFDTTH(3) LNRTPSLPSVHA(1) LQGSFSIHGNPP(1) TAGKVTASLIGR(1) VLGTKWPPMPLS(1) DHASTWMVKRGV(1) SSLPTPSESPSR(1) 7 Phage display (competitive panning) 3 PMID:16517013 UDP-N-acetylmuramoylalanine--D-glutamate ligase D-glutamate Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage encoding peptides were eluted in the each round of biopanning using glycine-HCl, glycine-HCl and D-Glu. 254 CKGTINPFC(45) CNVHRGLHC(16) CKLTANPTC(2) CXGAINPFC(1) 4 Phage display (common panning) 7 PMID:16882546 Trisialogangliosides (GT1b), NGF-differentiated PC12 cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In the present study, we have designed a biopanning strategy using two tiers of selection: the first on GT1b, and the second on NGF-differentiated PC12 cells. We have also combined different strategies to recover bound phage. In the first tier of biopanning, acidic elution (Glycine-HCl-pH 2.2) was used to capture all GT1b bound phage, then rTTC elution was used to select for phage with binding at the clostridial toxin receptor. In the second tier of cellular biopanning, PC12 cell lysis was used to recover all phage binding to or taken up by the neuronal cell line. Phage was amplified and titered after each round. 255 GRRTRSRRLRRS(7) GRRIAGPYIALE(5) TPRNLRTSNTHR(4) SMPINSPYIPWS(4) SMSIASPQIPWS(3) GRRINRLILPRN(3) GRRTRSSRLRNS(2) GRRPMKLNKTP(2) 8 Phage display (subtractive panning) 4 PMID:16458253 XGC9811-L cell line Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) High liver-metastatic cell variant XGC9811-L was chosen as a tester and its parental cells XGC9811 cell as a depletory. With a subtraction/selection protocol, multirounds of selection were carried out in our experiment. Before each round of selection, XGC9811 cells were used for preincubation with the library in order to remove the non-specific phages. Specific phages binding to liver-metastatic XGC9811-L cells were obtained from the phage peptide library and further enriched round by round. 256 CGRWSGWPADLC[56] 1 Phage display (common panning) 4 PMID:16252253 Integrin alpha-X-beta-2, integrin αxβ2 Intercellular adhesion molecule 1, ICAM-1 Not determined. Not determined. CL10 phage display library (CX10C) Surface plasmon resonance (SPR) The bonding phage was eluted with low pH. 257 CHKGHDRGKKRC(5) 1 Phage display (common panning) PMID:16252253 Integrin alpha-X-beta-2, integrin αxβ2 Intercellular adhesion molecule 1, ICAM-1 Not determined. Not determined. CL10 phage display library (CX10C) The bonding phage was eluted with EDTA-containing buffer. 258 MDKTHFVNE 1 Phage display (common panning) PMID:16252253 Integrin alpha-M-beta-2, integrin αmβ2 Intercellular adhesion molecule 1, ICAM-1 Not determined. Not determined. LL9 phage display library (X9) The bonding phage was eluted with low pH. 259 CPGGEWRSKAKC 1 Phage display (common panning) PMID:16252253 Integrin alpha-M-beta-2, integrin αmβ2 Intercellular adhesion molecule 1, ICAM-1 Not determined. Not determined. CL10 phage display library (CX10C) Bound phage were eluted with anti-CD11b/CD18 antibody CBRM1/29. 260 AHKSARKTE WSYWETVAK 2 Phage display (common panning) PMID:16252253 Integrin alpha-M-beta-2, integrin αmβ2 Intercellular adhesion molecule 1, ICAM-1 Not determined. Not determined. LL9 phage display library (X9) Bound phage were eluted with EDTA-containing buffer. 261 SSQVVGVPQLMQSSP(1) SAYAATVRGPLSSAS(1) DRVPLVHVIFNSFGY(1) RNQGPVKMVFPIAPS(1) EGQFTFPRGASE(1) 5 Phage display (common panning) 3 PMID:16546989 DNA topoisomerase 1 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) 262 AWPLSQLDHSYN YHLSSQQLDHSL ATWGHPRSSQGM QLDSH 3 Phage display (common panning) 3 PMID:16938891 Anthrax toxin receptor 1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 263 CPSSTLFAC 1 Phage display (common panning) 3 PMID:16938891 Anthrax toxin receptor 1 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 264 SPHGSTDHSTTA 1 Phage display (common panning) 3 PMID:16938891 Anthrax toxin receptor 2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 265 STDHSLY STDSGWV 2 Phage display (common panning) 3 PMID:16938891 Anthrax toxin receptor 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 266 CTSTDATYC 1 Phage display (common panning) 3 PMID:16938891 Anthrax toxin receptor 2 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 267 KSLSRHDHIHHH(5) 1 Phage display (common panning) 3 PMID:16953561 Hepatoma cell line SMMC-7721 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The KD (μM) was calculated and shown. 268 CLVDAAALC(8) CPIALGLKC(3) CGGPLKGLC(2) CINLGLTMC(1) CFSLGLIKC(1) CPAYKLYSC(1) CGSRSKGTC(1) CNSVGGRSC(1) 8 Phage display (common panning) 6 PMID:15306709 Plasmodium falciparum-infected red blood cells (iRBCs) Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library 269 KQRTSIRATEGCLPS(5) GRHRTSVPTDEVFIT(1) KKSHHPSSEWGLNLT(1) KQRDSRSGYTAPTLV(1) RNHGTDRATTIPPLS(1) 5 Phage display (subtractive panning) 3 PMID:15980377 Nontypeable Haemophilus influenzae, strain R2866 Not determined. Not determined. Not determined. f88-15mer phage display library (X15) To isolate peptides binding to strain-specific epitopes, we chose to preadsorb the starting library against a nonvirulent NTHi (Rd KW20) prior to affinity selecting for peptides binding to R2866. 270 PSALGRFTRGPL SLIFVTISSEWG LSLSLDLLTFRT PDIRHYFIQNRG GCRIVYRRPLHL RTAGFDIKLIDT RIQYQAISTVSL 7 Bacterial display (common panning) 5 PMID:15895468 Anti-sperm polyclonal antibody Sperm Not determined. Not determined. FliTrx bacterial display library (X12) 271 CEVSHPKVGC(4) CRARGQGWC(3) CRPYTGWKEC(2) CVGVGGTIPC(2) CIRGVARDSC(2) CEPEIRSNNC(2) CRVCRTWVLC(2) CWVTTSNQWC(1) CSGGSNRSPC(1) CKTIPSAATC(1) CTE*RKRRIC(1) 11 Phage display (common panning) 3 PMID:16397382 Newcastle disease virus Not determined. Not determined. Not determined. pSKAN8-HyA phage display library 272 TSNYSILYTEFA WDLTYELDRLWT ANLINEFDDLAS ESRLTSEFDGIH 4 Phage display (common panning) 3 PMID:16183141 Anti-NS4A monoclonal antibody 2E3C2 Non-structural protein 4A, NS4A Not determined. Not determined. Ph.D.-12 phage display library (X12) 273 TVLTNPSTNHLS(2) AANMNMSRVSHT(2) TACHQHVRMVRP(1) STAELHFPMVFP(1) ASPPSYALPVTP(1) APHHTNITEIRI(1) 6 Phage display (subtractive panning) 3 PMID:16598852 Hepatocellular carcinoma cell lines BEL-7402 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After the first round of panning, the phages were treated with normal liver cell line HL-7702 in the same way for reverse absorption, and the unbound phages were collected to eliminate the phages which can bind to liver cells. The residual phages were amplified and tittered for next round panning. After three rounds of panning and two rounds of reverse absorption the peptide sequences of randomly picked phage clones were analyzed by DNA sequencing. 274 NGNNVNGNRNNN 1 Phage display (subtractive panning) 4 PMID:16455333 Monoclonal antibody CM22 IgM Antigens exposed in ischemic tissue Not determined. Not determined. Ph.D.-12 phage display library (X12) The library was biopanned 4 times with the pathogenic CM22 IgM clone and then negatively selected against the inactive CM31 IgM clone. 275 GPGVSSAPPFSK LKSSGSAPPGPF AGKASKIPDPGF VPLSAGAPPLMA 4 Phage display (common panning) 3 PMID:16934463 Silver nanowires Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The generated binding peptide sequences indicate that both serine and proline residues are important for binding to AgNW, and the obtained sequences bear strong resemblance to the sequences previously obtained against AgNP. Taken together, these observations suggest that amino acid residues may be binding to the Ag atoms, and is indiscriminate of their structural morphologies. 276 NLLPTYTPLKLM HSNLPTKRPTSL CPNGLLGPCPSL KVWSLEPPGPAA TQPLGLLPSRHL IPHHHTLNMESH KPPSTWPQNALH LETRTTPPAKSQ KLVDESSTSPLS AGMATSRSTSPL KHVPVHSALSVN LEPQINSVGMVR 12 Phage display (common panning) 3 PMID:16414071 Protein tonB Ferrichrome-iron receptor 2GRX, Not determined. Ph.D.-12 phage display library (X12) 277 CILAALSAC CLPTGGLLC CLTRSPAAC CKSAFLPWC CTKPLTQQC CLAALKSTC 6 Phage display (common panning) 4 PMID:16414071 Protein tonB Ferrichrome-iron receptor 2GRX, Not determined. Ph.D.-C7C phage display library (CX7C) 278 TMGFTAPRFPHY KLVDESSTSPLS 2 Phage display (common panning) 3 PMID:16414071 Protein tonB Vitamin B12 transporter btuB 2GSK, Not determined. Ph.D.-12 phage display library (X12) 279 CTKPLTQQC CTGPLPNRC CSPRTTPFC CMLEKPRLC CNQPRGPQC CLTQTPTRC 6 Phage display (common panning) 4 PMID:16414071 Protein tonB Vitamin B12 transporter btuB 2GSK, Not determined. Ph.D.-C7C phage display library (CX7C) 280 KVWSLEPPGPAA AALGTYSTHTPT SSMKVWSLPPAP NLLPTYTPLKLM NNSQKPAPVSPF GNSVNKTWTHDY KHVPVHSALSVN 7 Phage display (common panning) 3 PMID:16414071 Protein tonB Fe(3+) dicitrate transport protein fecA Not determined. Not determined. Ph.D.-12 phage display library (X12) 281 CLTQTPTRC CHATLPPTC CSWDPAPLC CTLSPKLHC 4 Phage display (common panning) 4 PMID:16414071 Protein tonB Fe(3+) dicitrate transport protein fecA Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 282 SLKNYPVSWKNT NLLPTYTPLKLM TQPLGLLPSRHL HSNLPTKRPTSL QSPVNHHYHYHI KLVDESSTSPLS SHSNTTQTRPSD 7 Phage display (common panning) 3 PMID:16414071 Protein tonB Ferrienterobactin receptor Not determined. Not determined. Ph.D.-12 phage display library (X12) 283 CTGPLPNRC CSWDPAPLC CSHFAPHQC CSQVPLKSC CHMSPLGAC CMHMAPLSC 6 Phage display (common panning) 4 PMID:16414071 Protein tonB Ferrienterobactin receptor Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 284 INVPMMVDGITM(2) RGYPVVDWIFME(1) INIDGVAMDIGM(1) 3 Phage display (common panning) 2 PMID:16414054 Anti-tenascin-C monoclonal antibody ST2146 Tenascin,TN Not determined. Not determined. X12 phage display library 285 WHWLQWPQTPQP WHWQWTPWSIQP WHWSLWRPPYTL WHWTNWGRTSPA WPWWKNPNPNPT VHWWWNPPPPPI FHWPYYPAYWAL YPWHWWHSVSPW HNYRIWNHWWLS W-H-W-x(0,2)-W 9 Phage display (common panning) 3 PMID:15788194 Anti-MC-LR monoclonal antibody MC10E7 Microcystin-LR, MC-LR Not determined. Not determined. Ph.D.-12 phage display library (X12) 286 CQFSAQSRC CLATSGQPC CTNQPRSFC CQPHPYYMC CSLSLTRHC CLTNSYNTC CSEMRLALC CPEHNLRFC 18 Phage display (common panning) 3 PMID:15788194 Anti-MC-LR monoclonal antibody MC10E7 Microcystin-LR, MC-LR Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 287 ASPVTARVLWKASHV VSSLVLLSHGGPHSS IMVLCPLWLWLGTTS AVAHVTSRVPRWSAA 4 Phage display (common panning) 3 PMID:16290070 Anti-dsDNA polyclonal antibody Deoxyribonucleic acid, DNA Not determined. Not determined. f3-15mer phage display library (X15) 288 MSTRPPT(2) LAHTTQP(1) LTLQPLS(1) MQPWSLA(1) LATIPAS(1) LAKFPVA(1) LGKFPYP(1) LAKQPPR(1) LATLPKN(1) LTVLPQN(1) LASLPRV(1) LSLPSLY(1) MSLPLLK(1) MSLPFSG(1) MSTPPVT(1) MSKLPFN(1) MSALPSA(1) MQILPAS(1) MAVPTSI(1) LARTLTT(1) LQTFQNV(1) MASSVST(1) MAISPSS(1) MQIPTGY(1) MAAPTGY(1) MAAPIEH(1) MAIPRAA(1) MAHPGVS(1) MASPLFS(1) LASPPFY(1) FSMRIPI(1) MTIPITP(1) MGQPISP(1) MTTPQIR(1) MTMPVMA(1) MTLPPEA(1) LSRPPAA(1) MTLPMDH(1) FSRPAIA(1) LSHNPAP(1) LSHIPET(1) MQLLPPP(1) MTLTPPP(1) LTTPPRL(1) LTVPPAL(1) LTTPPFI(1) LTPPPRV(1) LTLPPRT(1) LTRPPMP(1) MASYGPG(1) LASRGPM(1) MASRTHP(1) MANRPHP(1) GTLLSRT(1) MTALPLR(1) MTSIPRN(1) LTWRMRS(1) MSTAWQL(1) MTTALSL(1) MTSATLS(1) YTSARLF(1) MTSTAPL(1) LTPVSVF(1) LTPMRTY(1) LQPPWLT(1) LTPSWLP(1) VPRLLPL(1) FAAMPAL(1) SNYNTQY(1) MYAIRTT(1) LSGPVKH(1) SGQAAVQ(1) LSRPPSQ(1) MTVPPSV(1) 74 Phage display (subtractive panning) 5 PMID:15949645 Endothelial protein C receptor Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated on four successive cultures of untransfected HepG2 cells in 1ml of biopanning media. Incubation of the library on untransfected HepG2s should effectively remove from the phage population those bearing peptides capable of binding receptors normally expressed on HepG2, thus clearing the phage library of phage able to bind to non-target receptors. Cleared phage library was then panned immediately in duplicate on HepG2-EPCR. Cells were extensively washed, weakly associated phage removed by acid wash prior to recovery of high affinity phage isolated by cell lysis. Four additional rounds of panning were performed including a clearing step on HepG2-beta-gal prior to biopanning on HepG2-EPCR. 289 WHWSLWRPPYTL(4) FHWRWPPSLQIM(2) FHRTPWLQFSLP(1) LHRPHLYHVWQR(1) WHWSWIQNAAPN(1) WHWSYASQLLQI(1) 6 Phage display (common panning) 4 PMID:15963509 Pesticidal crystal protein cry11Aa Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 290 CDGGWLSKGSWC[0.876 ± 0.078] CSGAWLSSGRWC[1.177 ± 0.054] CDPRALDYWLRC[1.045 ± 0.050] CVGDARLWWRDC[1.012 ± 0.022] CTSWLNEGRASC[0.887 ± 0.045] CGRLKMVPDLEC[1.219 ± 0.035] CGRLRMVPDLEC[1.166 ± 0.066] CIRESGMSAGEC[0.788 ± 0.032] CIRGFAMSAGDC[1.094 ± 0.038] CIRSSPLLPGDC[1.445 ± 0.030] CLRGDSLTPGDC[1.324 ± 0.128] CVRTSSMVPGDC[0.981 ± 0.087] CDPGWLSKGTWC[0.989] 13 Phage display (common panning) 3 PMID:15589320 Anti-GD2 monoclonal antibody ch14.18 Disialoganglioside GD2 Not determined. Not determined. CL10 phage display library (CX10C) ELISA Optical density was quantified in an ELISA reader (Dynatech, Denkendorf, Germany) at 405–409 nm. Data shown were reproduced from the graph. 291 PRRIKPRKIMLQ(53) 1 Phage display (common panning) 6 PMID:15919139 Anti-tumor monoclonal antibody BAT Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 292 QRILQQINLPRI(31) 1 Phage display (common panning) 3 PMID:15919139 Anti-tumor monoclonal antibody BAT Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the screening, the phage display peptide library was subjected to three rounds of affinity selection. Each affinity selection was done by biopanning on Petri dishes coated with streptavidin. Petri dishes coated with streptavidin were blocked with phosphate buffered saline (PBS) containing 3% bovine serum albumin (BSA) and 0.1 mg/ml streptavidin. 293 SHGFSRHSMTLI SVSVGMKPSPRP AFSNTHRMMPLK NHTTELGSLMPG 4 Phage display (common panning) 5 PMID:16029812 Human pancreatic adenocarcinoma cell line Capan-2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 294 PKWRHRY FNENWPR WDGNYTL RWRSNEP 4 Phage display (common panning) 3 PMID:15757647 C-C motif chemokine 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 295 HTRMHWHHVHTL HNWKHTHLKTPA SLWHGHKHHSPL HNWHWHSIRGRR SWKHHNFHSRYS WPHHHRSQRVAS KRFHPMHHTIRH HRPHYHTDYLTA SWPHHHRMPRLA 9 Phage display (common panning) 3 PMID:15757647 C-C motif chemokine 2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 296 AWLPWAK(4)[0.384 ± 0.010][272] NLQEFLF(2)[0.376 ± 0.022][NT] WMKPIW(1)[0.214 ± 0.010][NT] TYFGDWF(1)[0.225 ± 0.036][NT] MPPLWWS(1)[0.192 ± 0.010][NT] FVRPFAL(1)[0.186 ± 0.024][NT] 6 Phage display (common panning) 5 PMID:15721308 Lipopolysaccharide, LPS Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA,Surface plasmon resonance (SPR) Phage clones were allowed to react with beads conjugated with LPS extracted from S. enteritidis. The beads were exposed to horseradish peroxidase conjugated anti-M13 antibody and then ABTS substrate solution sequentially with an intermittent washing step. M13KO7 helper phage was used as a negative control. Unconjugated epoxy beads were employed to determine the background signal level. The optical densities were measured at 405 nm. Data shown are mean values of triplicate experiments and their standard deviations, and reproduced from the graph. Besides, the binding characteristics of the phage displaying AWLPWAK for the LPS of S. enteritidis were examined by surface plasmon resonance (SPR) analysis. The calculated kon, koff, and KD (KD=koff/kon) values were 1.68e6 1/Ms, 0.0457 1/s and 272 μM, respectively. NT, not tested. 297 HLNILSTLWKYR(35) TTVYPARWGAHP(3) SYQLSTHRWPLH(2) NHVHRMHATPAY(2) 3 Phage display (common panning) 4 PMID:16023583 Trisialogangliosides (GT1b) Tetanus toxin Not determined. Not determined. X12 phage display library Peptides specific for GT1b were isolated using a four-round biopanning strategy. The initial round of biopanning was carried out in triplicate, eluting bound phage with a nonspecific acidic elution. This stringent elution was performed to prevent the loss of strong-binding phage clones in early rounds. Also, to optimize the capture of GT1b binding phage clones, eluates from the three separate biopanned wells were combined and amplified for use in the second round of biopanning. In the second through fourth rounds of panning, the bound phage was eluted with the tetanus toxin C fragment (rTTC) to specifically isolate phage peptides that mimic tetanus toxin binding properties. 298 CEHGAMEIC CGGGRPASC CHLGVEHLC CEEKEPERC 4 Phage display (common panning) 1 PMID:15707681 Anti-EGF monoclonal antibody EGF-10 Pro-epidermal growth factor, EGF Not determined. Not determined. CX7C phage display library 299 DHYIYEEIY(9) DDFEYSYHN(1) DPDIYDYWY(1) DLGYYYFGR(1) NPDIYEEIY(1) 5 Phage display (common panning) 3 PMID:14715298 Src SH2-GST fused protein Not determined. Not determined. Not determined. X4YX4 phage display library 300 ELDEYMEYE(5) KLAAYFEYL(2) SQVCYSGYP(2) PIRCYARGS(1) PQSSYVAIS(1) VDYVYVSSE(1) 6 Phage display (common panning) 3 PMID:14715298 PTP-alpha-GST fused protein Not determined. Not determined. Not determined. X4YX4 phage display library 301 ELDEYMEYE(5) PVRCYVRGS(2) RIPVYVDAR(2) SIDDYVYGL(2) RIPVYCDAS(1) 5 Phage display (common panning) 3 PMID:14715298 PTP1B-GST fused protein Not determined. Not determined. Not determined. X4YX4 phage display library 302 QPSYTVPIPRHA(10) QPSYTVPIARHA(1) QPSYTVRIARHA(1) SPSYDIRVPRGW(1) DSSPDYTFTSAR(1) 5 Phage display (common panning) 4 PMID:15530735 Anti-gE monoclonal antibody 14B11 Envelope glycoprotein E, gE Not determined. Not determined. Ph.D.-12 phage display library (X12) 303 TVFPPWHLLNNA(5) SVSVGMKPSPRP(4) SVSVGMKPRPRP(2) SVSAGRKPRPRP(1) TVFPAWHLLNNP(1) 5 Phage display (common panning) 4 PMID:15530735 Anti-gE monoclonal antibody 13E8 Envelope glycoprotein E, gE Not determined. Not determined. Ph.D.-12 phage display library (X12) 304 YEPWWAGKVRHL(8) SVSVGMKPSPRP(5) SVPVGMKPRPRP(1) SGWSHLYWWTHD(1) 4 Phage display (common panning) 4 PMID:15530735 Anti-gE monoclonal antibody 17C9 Envelope glycoprotein E, gE Not determined. Not determined. Ph.D.-12 phage display library (X12) 305 SVSVGMKPSPRP(2) SHHMQHGPWMFV(2) APTHWCYYPPLI(2) SVSVGKKPSPRP(1) SVSVGLKPSPRP(1) SDQLLLLSSANW(1) SVSVGMKPRPRP(1) ACDQMECYPPLV(1) ASDQMECYPPLV(1) APTHWFYYPPLI(1) APTHWYYYPPLI(1) 11 Phage display (common panning) 4 PMID:15530735 Anti-gE monoclonal antibody 8G11 Envelope glycoprotein E, gE Not determined. Not determined. Ph.D.-12 phage display library (X12) 306 SGFDPLITHA(3) SDWDPLFTHK(2) MAEVLENSVG(2) MAEVLQKESG(1) MAEVKLNSSG(1) TLKFVDSWGL(1) 6 Phage display (common panning) 4 PMID:15033456 Amino acid 27-135 of SLAM Hemagglutinin glycoprotein Not determined. Not determined. X10 phage display library 307 CSRLGRSSAWVC(9) CPFWVRGTTWC(2) CRLTENTEPLLC(2) CTHWQLGERPDC(1) CPSTPGERVRHC(1) CRGGPDDLTALC(1) CPSTPGSRQNMC(1) CPSTPGDNPLVC(1) CKFVVNGRWIDC(1) CKFLVNGRWIDC(1) CFTWGGLRDKSC(1) 11 Phage display (common panning) 3 PMID:15577826 Anti-phl p 5a monoclonal antibody IgE Pollen allergen Phl p 5a Not determined. Not determined. CL10 phage display library (CX10C) Bound phages were eluted with glycine buffer. 308 CERAGAMERANC(22) CRSVSKEEPGMC(1) CKLGKFGAARVC(1) CVQDLMKSSGVC(1) 4 Phage display (competitive panning) 3 PMID:15577826 Anti-phl p 5a monoclonal antibody IgE Pollen allergen Phl p 5a Not determined. Not determined. CL10 phage display library (CX10C) Bound phages were eluted competitively by using recombinant Phl p 5a in Hepes/NaCl. 309 YSGSWWPPTYNNEVPL HTWWYNPPSYMGLEAS GTWTWWPPTYAGMDHL TLWGLFPPVYEDSFGL PWTSWWPPVYEGSTTN 6 Phage display (common panning) 3 PMID:15135114 Long neurotoxin 1 Not determined. Not determined. Not determined. X6PPX(Y/F)X6 phage display library 310 CSISTHSWC[3.32] CYGNWFSNC[2.65] 2 Phage display (common panning) 3 PMID:14736416 Anti-MBP monoclonal antibody MK16 Fab Myelin basic protein, MBP Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance was measured at 490 nm using an immunoreader (Emax). Data shown were reproduced from the graph. 311 CNYKTHSWC[3.76] CNYSVAHLC[3.76] CKWSVAHLC[2.12] CSIKTHSWC[NT] 4 Phage display (common panning) 6 PMID:14736416 Anti-MBP monoclonal antibody MK16 Fab Myelin basic protein, MBP Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance was measured at 490 nm using an immunoreader (Emax). Data shown were reproduced from the graph. NT denotes not tested. 312 SLNDWIDWSEPH SLNDWIDWAEPH LTVRSDTTKYSL SHSLMTSSSVWT EFIKSGRTLDRL SHTTPGKNNDPF SMHSFGGESWSQ LPG(M/L)KHTPSPTD SSNLPTAQALLK TLSHQNMDQRSN FPGVVVRPALGQ DTRPMTHLLEPV NYARDPLMSLPQ NNSYSPVLAYRV STAQKNVPTGLQ HLFKSNTLL(H/D)PP SLNDWIDWADPH SPTMPSSFALTE NTHHLLWKLARA SHHTQSFMQKTQ TNAPPPLLSSWA NWEKQIGWPSAS NSPWPSWSLPFR TGAFSNPYMFRN SPMKGTPTWSPN TSRAEASITRVF SVSVGMKPSPRP HNMWIPMLRASH SVSVWMKPSPRP SQAIQNLPGPG AYPFQVTRQPAG AEPFPSPKGSLK TQWNMYNPWTRQ SVSGGMKPSPRP SVSVGMKPSHRP GGIACTKWSPQG IANKGPPSHAAP TYAQLQPTPSLR ANPMKYLLSRPL GPTMRWTIPTPF GSFASTARMPGL SHKPPTNVIARL 42 Phage display (common panning) 5 PMID:15325109 Cerebellar granule neurons Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 313 WAPPLFRSSLFY(4)[32] NFMESLPRLGMH(2)[19] FYSHSFHENWPS(2)[9.6] FHENWPSGGGSA(2)[3.4] 4 Phage display (common panning) 3 PMID:15279884 Self-assembled monolayer (SAM) of a Gb3 mimic Shiga toxin Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR) The dissociation constant, KD (μM), was calculated based on the SPR evaluation software. 314 LRRDYIRSP(2) QRRAATARS(1) ARRRAEPML(1) RKRSSADPL(1) RRHKSAPSS(1) KKYRNMAGP(1) FRAARHPQL(1) IRRLSGGPP(1) TPRLPGALE(1) WKKKLGASP(1) CTRGYLERRLC(1) 11 Phage display (common panning) 3 PMID:15163538 Anti-collagen alpha-1(II) chain monoclonal antibody CB268 Collagen alpha-1(II) chain Not determined. Not determined. pVIII-9aa and pVIII-9aa.Cys phage display library pool 315 RRLPFGSQM(4) RYAFGSQIA(2) HRLAFGQNT(1) HRLAFGQYT(1) HEHTFGRQW(1) RAAPFGNQW(1) TRAFGNEAT(1) RRLPFGSSL(1) TRSFGIQAT(1) CIAPKRHNSAC(1) CESAQRPFGCC(1) 11 Phage display (common panning) 3 PMID:15163538 Anti-collagen alpha-1(II) chain monoclonal antibody CII-C1 Collagen alpha-1(II) chain Not determined. Not determined. pVIII-9aa and pVIII-9aa.Cys phage display library pool 316 NRKDWIERTL(10) 1 Phage display (common panning) 3 PMID:15680153 Anti-nucleosome monoclonal antibody #32 Nucleosome Not determined. Not determined. L100 phage display library (X10) 317 EYRARGWGGWCR(2) 1 Phage display (common panning) 4 PMID:16399397 GST A1-1 fused protein Not determined. Not determined. Not determined. CX9C and CX15C phage display library pool 318 CGGGVWAVSGC(6) 1 Phage display (common panning) 3 PMID:16399397 GST P1-1 fused protein Not determined. Not determined. Not determined. CX9C and CX15C phage display library pool 319 CGVVRGPSRGC(5) 1 Phage display (common panning) 3 PMID:16399397 GST M2-2 fused protein Not determined. Not determined. Not determined. CX9C and CX15C phage display library pool 320 AQAFDYVPAWVV GFSYYRPPWIL LSPYRPSWVNLVGTS AVAYPVSAYVPAWVS AVVAVPSYIPTWIG YHACPPWVGIHRCEL VGLGDYGLDFAYVS GVMLCPQYFPSCRSL SVYALPDLSVPVWVR VHMIKPPWVGRLSSG ESFVPPWVVTVPPPR APGRVPALLIPPWAW GLFVPPFWGTTSGDL 13 Phage display (common panning) 3 PMID:14871530 Anti-MCPS monoclonal antibody 1E4 Meningococcal serogroup C capsular polysaccharide, MCPS Not determined. Not determined. f3-15mer phage display library (X15) 321 SYQWWWHSPQTL(10) WHQTYTSSLWES(6) HSSWYIQHFPPL(5) HSSPYTDYLWAV(3) 4 Phage display (common panning) 5 PMID:15522450 psi RNA Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 322 HPPMNASHPHMH HTKHSHTSPPPL HVSHFHASRHER HLASGHSIHYRT HQAHNHTHPSSL HGSKANHPHIRA HTPSNHRHTHNW HAPHTHMRSWSA HVSHHATGHTHT HKLPSASRHHFH HTTPSHLHPHSR DPSTHQHPPHKH SPQHTHHARIKN HINHHHDTPSYR HPGVHSHPSPTP TVVQTYSMVTRA FSYRQSPPPPLY IMQNSISSPEML 18 Phage display (common panning) 3 PMID:16257264 Silicon dioxide, SiO2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 323 WHRTMWWWP(8)[0.75 ± 0.03] KPWWRTNTA(6)[0.99 ± 0.01] QNTDWLGNL(3)[1.06 ± 0.11] GMKAVRIQG(1)[1.44 ± 0.14] GKAMLDRAS(1)[1.60 ± 0.01] SHATVKAAV(1)[NT] PWWWMTRHW(1)[NT] HARSHYPWY(1)[NT] KWKWPDRPK(1)[NT] SLEHRAFRN(1)[NT] GKLNLGGYK(1)[NT] KMNKGVVNP(1)[NT] 12 Phage display (common panning) 6 PMID:10772861 Laminin-1 Not determined. Not determined. Not determined. J404 phage display library (X9) ELISA ELISA measurements of the binding to laminin-1 expressed as A405. Wild-type M13 phage was used as a control. Data shown were reproduced from the graph and expressed as the average of replicate wells ± SD. NT denotes not tested. For the first three round of panning, Bound phage were eluted using HCl-glycine. In rounds 4, 5 and 6, we applied a consecutive specific elution at neutral pH, which consisted of an elution with peptide 11 (CDPGYIGSR) in TBS. 324 DRTAMQVAA(2)[1.18 ± 0.01] VVSKISEAG(1)[NT] GGSVAFRAG(1)[NT] GPGAWWGSA(1)[NT] SKMHRNSWF(1)[NT] AKIPAGRDR(1)[NT] KMNKGVVNP(1)[NT] 7 Phage display (common panning) 6 PMID:10772861 Laminin-1 Not determined. Not determined. Not determined. J404 phage display library (X9) ELISA ELISA measurements of the binding to laminin-1 expressed as A405. Wild-type M13 phage was used as a control. Data shown were reproduced from the graph and expressed as the average of replicate wells ± SD. NT denotes not tested. For the first three round of panning, Bound phage were eluted using HCl-glycine. In rounds 4, 5 and 6, we applied a consecutive specific elution at neutral pH, which consisted of an elution with mixture of heparan sulfates from bovine kidney and bovine lens capsule in TBS. 325 STLSGFDLLGAI YMTSSSGPSRFS 2 Phage display (common panning) 3 PMID:15001356 Heat shock protein HSP 90-beta, HSP 90 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 326 TQLATDFGSRLP AYNMSTLARDFH HTNMSQMLHSGD HNNMSQMLHSGD HVNTSVGMLRHP 5 Phage display (common panning) 3 PMID:15001356 E3 ubiquitin-protein ligase NEDD4 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 327 WSTSEHFKYYAM AHRSDFWRPFPT AVATDLWATESE IPRNQDSYLLRTH 4 Phage display (common panning) 3 PMID:15001356 Death-associated protein kinase 3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 328 SHPMHEFNRTHI(2) VSFQELWAHLAL(1) TKNFLSYWEELG(1) SHDPTHWHSSRR(1) TSFAEYWNLLSG(1) LPFSDYWRALCE(1) KVWTLNPDHPPV(1) TWLDNIWTTLGP(1) 7 Phage display (common panning) 3 PMID:15001357 E3 ubiquitin-protein ligase Mdm2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 329 QHWLAQR(1) QHWAIHN(1) QHWAHSM(1) EHWTWPV(1) WHWTSAT(1) WHLTKPT(1) APWAWYP(1) LAPWNSD(1) FHFTRLD(1) HHWVMSD(1) HHWASSN(1) HHWTSSN(1) THWASLR(1) THWAHDS(1) THWTGDP(1) SHIYRTS(1) NHWASAEPLDVV(1) 17 Phage display (common panning) 3 PMID:15914189 Anti-CCR5 monoclonal antibody 3A9 C-C chemokine receptor type 5 Not determined. Not determined. Ph.D.-12, Ph.D.-7 and Ph.D.-C7C phage display library pool 330 CHASIYDFGSC(18) CVYALIMPPLC(1) 2 Phage display (common panning) 5 PMID:15914189 Anti-CCR5 monoclonal antibody 3A9 C-C chemokine receptor type 5 Not determined. Not determined. pVIII-9aa and pVIII-9aa.Cys phage display library pool 331 STALPGL(6) TATTWTG(2) TWHDFPL(1) GKWAPLF(1) SALFMTP(1) IPLNTTM(1) VQGPWPM(1) WVQTPLT(1) SAQDWWQ(1) RPPAIYP(1) TVEHTPF(1) NISNEVY(1) SHMFATP(1) ELARQNP(1) VDLLPRE(1) TMLLPTK(1) FPWHTQS(1) NHSKAFW(1) APERSPQ(1) KLVTPSS(1) NGLRLVT(1) 21 Phage display (common panning) 3 PMID:15914189 Anti-collagen alpha-1(II) chain monoclonal antibody CII-C1 Collagen alpha-1(II) chain Not determined. Not determined. Ph.D.-12, Ph.D.-7 and Ph.D.-C7C phage display library pool 332 RRLPFGSQM(4) RYAFGSQIA(2) RRLPFGSSL(1) RAGRFGYQR(1) TRSFGIQAT(1) SRLAFGDQL(1) HEHTFGRQW(1) TRAFGNEAT(1) RAAPFGNQW(1) HRLAFGQNT(1) HRLAFGQYT(1) CIAPKRHNSAC(1) CESAQRPFGCC(1) 13 Phage display (common panning) 5 PMID:15914189 Anti-collagen alpha-1(II) chain monoclonal antibody CII-C1 Collagen alpha-1(II) chain Not determined. Not determined. pVIII-9aa and pVIII-9aa.Cys phage display library pool 333 WTDMFTAWWSTP(4) GLWRFWFGDFLT(3) GGWTQWWWTAFY(1) NNWWYWWDTLVN(1) QMMLTLLWAFWY(1) MSEALWTAWTQW(1) MTGRLISWWWSL(1) WVEYMYSWIPTA(1) 8 Phage display (common panning) 5 PMID:17192308 PreS domain Not determined. Not determined. Not determined. X12 phage display library pThioHis-PreS and pTXB1-PreS were constructed for the production of thioredoxin-fused PreS (Thio-PreS) and chitin binding domain-fused PreS (CBD-PreS) in Escherichia coli, respectively. In the first two rounds and the last round, microwells were coated with Thio-PreS overnight in bicarbonate and saturated with 10% nonfat milk in phosphate-buffered saline (PBS). CBD-PreScoupled chitin resins were used in the third and the fourth rounds of screening. 334 MSPRATI(6) KLMTHWP(1) GLSLPPG(1) LPDTLSS(1) QPPLTLN(1) 5 Phage display (common panning) 3 PMID:16916959 Anti-LAM monoclonal antibody CS-35 ManLAM Not determined. Not determined. Ph.D.-7 phage display library (X7) 335 SHRLLQTYWSSA(8) YMDTQTTLPIMW(1) 2 Phage display (common panning) 3 PMID:16916959 Anti-LAM monoclonal antibody CS-35 ManLAM Not determined. Not determined. Ph.D.-12 phage display library (X12) 336 EQPYLHV(6) MSPRATI(4) EQPYIEN(2) SMITDLL(2) SMMTELL(2) MPFVTHN(2) SMIRDLL(1) EQPYRSM(1) NLTDINL(1) NLTDILP(1) TMDLGRF(1) YMDLGMK(1) LSQNASV(1) PAPNASL(1) AVQGFNW(1) ATHFMRI(1) KPSDFPP(1) QPLHSPL(1) 18 Phage display (common panning) 3 PMID:16916959 Anti-BCG polyclonal antibody Mycobacterium bovis BCG Not determined. Not determined. Ph.D.-7 phage display library (X7) 337 GWRSLDDSG(7) GPHSFDDSG(4) GRQSYDDSG(1) GRWSIDDSG(1) GTLSMDDSG(1) GHRSIDDSG(1) G-x(2)-S-x-D(2)-S-G 6 Phage display (common panning) PMID:15585859 Anti-p22 monoclonal antibody NS1 Cytochrome b-245 light chain Not determined. Not determined. J404 phage display library (X9) 338 WSPPEPKRR(20) SDWSPPNPR(1) FWTPVEPRR(1) YPIWTPITV(1) ELWTPVQVP(1) 5 Phage display (common panning) PMID:15585859 Anti-p22 monoclonal antibody NS2 Cytochrome b-245 light chain Not determined. Not determined. J404 phage display library (X9) 339 KIFGPFDFG RVHPFEMTSG GGKGNPFDF KTGPFDFDH NKLGPFDLT LGHPFDLIW KKVGPFDFG KGHPFDLIW KGYIFDIHL KHHPFELEN 10 Phage display (common panning) PMID:15585859 Anti-p22 monoclonal antibody NS5 Cytochrome b-245 light chain Not determined. Not determined. J404 phage display library (X9) 340 RGWISPIPG YPVWVNPIP RWINPIPWV INPIPRGLW RDNWINPIP RVGWISPIP VNPIPFNAW DTGWVNP 8 Phage display (common panning) PMID:15585859 Anti-p22 monoclonal antibody CS6 Cytochrome b-245 light chain Not determined. Not determined. J404 phage display library (X9) 341 AGGRSVNPS AGTYWVNPI GRFAVNPYM SEVWRMNPV FDLSRMNPE QWRDLMNPI SLLYEINPG VCIVCMNPE GVLLEMNPY ASWDVNPVP QWWDIMNPN 11 Phage display (common panning) PMID:15585859 Anti-p22 monoclonal antibody CS8 Cytochrome b-245 light chain Not determined. Not determined. J404 phage display library (X9) 342 KEAPKKLTE(2) SFMPKKNSE(2) TEARKKISE(1) PFMMKKPSE(1) KMTYKKPSE(1) AFMMKKPSE(1) KLMQRKPSE(1) KMRAIKPTE(1) KENTWFKPS(1) ANFMKKKEP(1) 10 Phage display (common panning) PMID:15585859 Anti-p22 monoclonal antibody CS9 Cytochrome b-245 light chain Not determined. Not determined. J404 phage display library (X9) 343 ALLRYNPFYYLSFSP(20) 1 Phage display (subtractive panning) 4 PMID:16280164 C-C chemokine receptor type 5, CCR5 Not determined. Not determined. Not determined. CDX3KPCALLRYX10 phage display library The library was subjected to cycles of selection on CHO cells stably expressing the CCR5 receptor. To decrease the background of phages binding to other cell surface proteins, the phage library was pre-adsorbed on wild-type (CCR5-negative) CHO cells before each of selection cycle. 344 CVPRRWDVC(2) CVSNPRWKC(1) CQHTSGRGC(1) CRARGWLLC(1) CGGVHALRC(1) CFNRTWIGC(1) CSRGPAWGC(1) 7 Phage display (in vivo) 3 PMID:15681844 Mouse pancreas Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Phage recovery from laser pressure catapult microdissection (LPCM) islet sections was achieved by bacterial infection. 345 CWSRGQGGC(21) CHVLWSTRC(11) CLGLLMAGC(11) CMSSPGVAC(9) CLASGMDAC(8) CHDERTGRC(6) CAHHALMEC(5) CMQGAATSC(4) CVRDLLTGC(3) CMQGARTSC(1) CGGLVAVGC(1) 11 Phage display (in vivo) 3 PMID:15681844 Mouse pancreas Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Phage recovery from laser pressure catapult microdissection (LPCM) islet sections was achieved by subcloning nested PCR-amplified peptide insert sequences into the fUSE5 phage genome. 346 SWKLPPS QPLLKLP HSTSPTA VPPQAPL AQIPPAS ANPPLPR FLPPLPS AAPLWAS 8 Phage display (in vivo) 5 PMID:16984380 Human gastric cancer cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 347 CDHMGIGSC CFRSGAPAC CGAIGSRGC CGRSGIRGC CHGRSGGRC CHRSGGRWC CISLGSFPC CMFSGASRC CRSAGSFMC CRSGFSDVC CSGAGVFAC CSGASAVPC CSGAWLGLC CSGRSGLIC CSRRSGEAC CTYRSGTRC CVPSGAKLC CVRRSGLNC CYAIGSFDC 19 Phage display (in vivo) 3 PMID:16581960 C57Bl/6 female mice muscle Not determined. Not determined. Not determined. CX7C fUSE5 phage display library 348 CDSRDLVSC CDVGLVSTC CEWSGLLYC CFGGRVSSC CFLVSAVRC CGNLSWSGC CHCCGWSGC CLVSLYGVC CPVSSGYXC CRTLVSSRC CSGFLWSGC CVLVSAAAC CVSSIAGCC CWSGPGPLC CWSGSAPRC CXXVSSXLC 16 Phage display (in vivo) 3 PMID:16581960 C57Bl/6 female mice pancreas Not determined. Not determined. Not determined. CX7C fUSE5 phage display library 349 CELGGNPDC CFLGGVRAC CKVERGFSC CLGSGRGFC CLRGFSDGC CQITTLGGC CSHWMLGGC CSLGGHIGC CWSLGGWRC 9 Phage display (in vivo) 3 PMID:16581960 C57Bl/6 female mice brain Not determined. Not determined. Not determined. CX7C fUSE5 phage display library 350 CAQLGSSSC CDRGMTNLC CERRVVSEC CFLGSCGRC CFLGSPFMC CGWGSDRGC CLLGSNWLC CLSLSKVPC CLSLSLRTC CNGGILGSC CQSLSISLC CSAVHLGSC CSLSGRRVC CSLSSSCGC CVDRGGGRC 15 Phage display (in vivo) 3 PMID:16581960 C57Bl/6 female mice kidney Not determined. Not determined. Not determined. CX7C fUSE5 phage display library 351 CARGSSQSC CASPGSSFC CCAIGGAAC CFYGAAELC CGAASVKIC CGAAYSALC CGDRLLGPC CGRWLLGGC CGSSLRSLC CGVGSSDRC CIGGGGSSC CIHGAAVYC CLLGGQLSC CLLGVGILC CRGSSAWAC CSVRSLLGC 16 Phage display (in vivo) 3 PMID:16581960 C57Bl/6 female mice uterus Not determined. Not determined. Not determined. CX7C fUSE5 phage display library 352 CAAGVGVNC CFAGVFVLC CTRAGVVTC CVAWRDSLC CLWRDRAGC CARDWRDYC CFSWRDARC CTSTFGGRC CACFGGEGC CVVLRFGGC CERGWFGGC 11 Phage display (in vivo) 3 PMID:16581960 C57Bl/6 female mice bowel Not determined. Not determined. Not determined. CX7C fUSE5 phage display library 353 VHPKQHR TASNNNS TISNKSQ TYSNSYP 4 Phage display (in vivo) 3 PMID:17000904 Vascular cell adhesion protein 1, V-CAM 1, VCAM-1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) An M13 phage library was injected into the tail vein of cholesterol-fed apoE-/- mice. Internalized phages were retrieved from dissected aortic plaques and then reinjected into subsequent mice for a total of 3 rounds of positive selection. 354 CVHSPNKKC[1.000] CDHASPMHC[0.574] CPTRIGQMC[0.141] CMHRAHQMC[0.370] CISHQMPAC[0.154] CPGHHTSQC[0.272] CNNAHHKNC[0.190] 7 Phage display (subtractive panning) 4 PMID:15653572 Vascular cell adhesion protein 1, V-CAM 1, VCAM-1 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance at 650 nm was measured (Emax; Molecular Devices). Data, relative absorbance, were reproduced from the graph and shown. Phage selection and negative depletion were performed using strain-matched murine cardiac endothelial cells (MCECs) and murine lung endothelial cells (MLECs). 355 CSSSPSKHC(8) 1 Phage display (in vivo) 2 PMID:16565728 Mouse abdominal skin Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 356 SWLELRP(7) EGWHAHT(5) KLWTIKP(2) 3 Phage display (common panning) 3 PMID:12649290 V-type proton ATPase 116 kDa subunit a isoform 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 357 WHLPFKC(6) WHKPFRF(3) HSLRTTF(1) LPKRSPI(1) W-H-x-P-F-x(2) 4 Phage display (common panning) 4 PMID:17401149 Vascular endothelial growth factor A, VEGF-A Vascular endothelial growth factor receptor 1, VEGFR-1 1FLT,1QTY, Not determined. Ph.D.-7 phage display library (X7) The recombinant human VEGF165 contained in bovine serum were used for screening. The template is likely to be VEGFR1; however, other possiblities exist. These peptides inhibit VEGF dependent human umbilical vein endothelial cell proliferation. Two selected peptides with sequences WHLPFKC and WHKPFRF bind the VEGF homodimer in a concentration-dependent manner, with micromolar affinity, and with a 2:1 peptide:VEGF stoichiometry. They inhibited HUVEC proliferation in vitro by 77 and 55%, respectively. 358 CAGALCY(28) CXRCTVLLACKL(2) CAGALCYYRAKC(1) CAGXAVTXGGK(1) CAGXPGXAG(1) CTGRMTXQXXXA(1) CQGNKTVQNSVS(1) CAATDHQPEAKCKLA(1) CATQVQHEAMHC(1) CTGXCDLXQR(1) YDPXKQL(1) CPAEXRAGPVTT(1) CEKNPPTVXNY(1) VTPGTVSLRPHS(1) FTPRTIGHINWC(1) CMVERKRDEARV(1) LYMIKWPGSEDC(1) CSXCMDXFWAFL(1) CAGALCY 18 Phage display (in vivo) 5 PMID:17377744 Mouse brain microvasculature Not determined. Not determined. Not determined. CX10C phage display library For each round, purified phages in PBS were injected into mouse via the tail vein. One hour later, brains were harvested, rinsed with cold PBS and homogenized for titering and amplification. This way, phages binding to brain microvasculature were enriched. The template of the brain-specific peptide CAGALCYA is surpposed to be a receptor or ligand involved in cell adhesion to endothelium of the brain microvasculature. Screening in vivo has inherent issues of high background, all the more apparent when a high complexity library is administered. The authors suggest there is an upper limit of library diversity of one in 10e5 pfu, if a hit is to be reliably identified by in vivo phage display. X in the mimotope sequence denotes unknown residue. The brain-specific peptide CAGALCY inhibits platelet adhesion to endothelium in vivo. 359 CFNGKDWLYC(9) 1 Phage display (common panning) 3 PMID:17973501 Phenoxibenzoic acid-polyclonal antibody complex Not determined. Not determined. Not determined. pAFF/MBP vector-based phage display library Antibodies specific for PBA, were purified on 3-((2-oxoethoxy)ethoxy)phenoxybenzoic acid coupled Sepharose, from serum of rabbits immunized with a hapten-KLH conjugate. The phage borne peptide did not bind to the analyte-KLH conjugate, which demonstrated that the peptide is specific for the PBA-antibody immuncomplex. 360 QPWLEQAYYSTF YPHIDSLGHWRR LLADTTHHRPWT SAHGTSTGVPWP VPWMEPAYQRFL TLPWLEESYWRP 6 Phage display (common panning) 4 PMID:17187899 Umbilical endothelial cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The biopanning is performed in parallel on three different groups of endothelial cells: (a) cells incubated under normal oxygen conditions; (b) after 3h of hypoxia; (c) after 24h of hypoxia. Peptides obtained did not induce VEGF receptor gene expression,indicating the template is not VEGF. The authors have isolated 40 individual phage clones from each group of endothelial cell screenings. A total of 15 non-identical sequences were obtained. However, only 6 unique sequences were given in the published paper. QPWLEQAYYSTF, YPHIDSLGHWRR and LLADTTHHRPWT originated respectively from endothelial cells under normoxia, 3 and 24h of hypoxia. SAHGTSTGVPWP was identified on cells under 3 and 24h of hypoxia. VPWMEPAYQRFL was found in both normoxia and 24h hypoxia cell groups. TLPWLEESYWRP is common to endothelial cells under normoxic and hypoxic conditions. These peptides induced angiogenesis as demonstrated by endothelial cell proliferation, migration and tube formation. Injection of peptides into the ears of mice resulted in increased numbers of blood vessels. They did not induce VEGF receptor gene expression indicating a possible VEGF unrelated mechanism. 361 VLIMPVLLGIPLLC(0.7) VXNCERITISRILN(0.1) VXNCERITISQNSKL(0.1) RSLNHASSFGYSVIM(0.1) 4 Phage display (common panning) 4 PMID:17959143 Proteinase K Protease inhibitors Not determined. Not determined. pComb3 X15 phage display library VLIMPVLLGIPLL inhibits proteinase-K activity with an inhibition constant of 4 μM. VLIMPVLLGIPLL revealed a considerable homology with a short peptide stretch of the Kuniz-type serine-protease inhibitor from Caenorhabditis elegans. 362 VWNCERITISRLIN(0.5) 1 Phage display (common panning) 4 PMID:17959143 Cathepsin S Protease inhibitors Not determined. Not determined. pComb3 X15 phage display library VWNCERITISRLIN showed functional inhibition of cathepsin-S induced sprouting of endothelial cells.Its IC50 is 13 μM. VWNCERITISRLIN showed homology with serine-protease inhibitor 8 from Rattus norvegicus. 363 CLRNSPRKQADRILNC(0.6) 1 Phage display (common panning) 4 PMID:17959143 Cathepsin S Protease inhibitors Not determined. Not determined. pComb8 CX15C phage display library The IC50 0f this peptide for cathepsin-S inhibition is 26 μM. 364 THALWHT(4) FDFWITP(3) SHALWHT(1) VPAVPLD(1) THALWHT 4 Phage display (common panning) 4 PMID:18040053 Hronchial epithelial cells (16HBE14o-) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 365 FDFWITP(7) FDFWLTP(1) FYALEFP(1) FDFWITP 3 Phage display (common panning) 5 PMID:18040053 EGTA-treated 16HBE14o- cell monolayers Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Before they were used to screen the library, monolayers of human bronchial epithelial cells (16HBE14o-) were treated with a calcium chelator, EGTA, to increase accessibility to the junctional complex/paracellular space. FDFWITP might be binding to the tight junction complex or other cellular components that are more available as a result of EGTA treatment.FDFWITP, identified as a potential tight junction modulator. 366 MMFKGQRVERVLT HNEFRQRETYMVF NWQEFQAKRSVAY LELKSNSVIMRWP DYNEVRRQMSMQM ALEMRAADVEYHF VEHLMEVQRKTTW GVEVKRQLSYHYM QELVGANIETYML QQMEVSRYVQYKW LQSFRQAPVDIWW QELRGKISIQPFK QQEYMSGQYDIIF SMEFAATVTSTFE EQQLKGRQTHIII MELKGQTDMFYII GAYAVGRWSYVDA GQFATSPKITIHK DVQEFRGVTAVIR HEARTVSTTYLML YMEMRGSTTVFFN QELIGSYSVMPTN HYYMEATRDIEMV NEAHSSGITIMLR DHPMEFRSKITMK TFAEMKGTVSYAL GVHMESMRRYTVI FQEYTGTYDIMDP FQAVEASKTLHFW YLETSRTYTTVWP TDYLEVRSQPIIY TFEQEVRAPNISW PQEVQGIAVEWV AEAKASTLHVYLM DYMEVVGNKISYI VIMEAVGRKTILQ FQAEAARAVTYSS EDYVYVKDVGTTN QEYKAHHSYKLMS YNEYRATPTFAVV EYFHANTTRIVQS ALEASRFISWDIN WEAVAAPIMHTWV FQELKAAETFWM NTLYAVAPPVIYV FQPYEVQRITTVM KPMESGRRTTVYY MEFKGALQYRLQP PQEVKQARKWIIE YRQQEVKRHIQIV E-[AFVLMI]-x(0,1)-[RK]-x(2,3)-[ST]-[VYIFWMLA] 50 Phage display (common panning) 3-4 PMID:17311924 A disintegrin and metalloproteinase with thrombospondin motifs 4 Not determined. Not determined. Not determined. X13 phage libray ADAMTS-4 was cloned and expressed in Drosophila Sf9 cells. The purified preparation was composed almost solely of the 70-kDa active form. The authors have identified the ADAMTS-4 cleavage motif E-[AFVLMI]-X(0,1)-[RK]-X(2,3)-[ST]-[VYIFWMLA], with Glu representing P1. Several 13-mer peptides such as DVQEFRGVTAVIR and HNEFRQRETYMVF, were shown to be substrates for ADAMTS-4. 367 CRKRLDRNC(8) CMNPKKQRC(4) CRSTKSSAC(2) 3 Phage display (common panning) 3-4 PMID:19012727 Human primary atherosclerotic tissues Interleukin-4, IL-4 Not determined. Not determined. CX7C T7 phage display library Primary human atherosclerotic tissues were obtained by endarterectomy of three patients with coronary atheroma (50 ± 12 year-old). Cell suspensions were prepared by homogenizing the tissues. Twenty-seven phage clones from coronary atheroma tissues were randomly picked and sequenced. However, only 3 unique sequences were given in the published paper. It has been demonstrated that the CRKRLDRNC peptide homes to atherosclerotic plaques through binding to IL-4R. CRKRLDRNC is a mimic of IL-4(P05112). According to the BLAST results, CMNPKKQRC may be a mimic of integrin β2 precursor(P05107); and CRSTKSSAC, Thrombospondin-4 precursor(P35443). 368 CPSNGKRDC(2) CFSEVD(2) CRQGGKRDC(1) CRTTRSKTC(1) CRTTQSNTC(1) CMHSEVD(1) CLIGEVD(1) 7 Phage display (common panning) 3-4 PMID:19012727 Human primary atherosclerotic tissues Interleukin-4, IL-4 Not determined. Not determined. CX7C T7 phage display library Primary human atherosclerotic tissues were obtained by endarterectomy of three patients with femoral atheroma (63 ± 11year-old). Cell suspensions were prepared by homogenizing the tissues. Twenty-seven phage clones from femoral atheroma tissues were randomly picked and sequenced. However, only 7 unique sequences were given in the published paper. The BLAST results indicate that the templates of the obtained peptides might include MMP-16(P51512), Scavenger receptor class A member 3(P35443) and Toll-like receptor 4 precursor(O00206). 369 THFATMF LQPTMHK FKPPASL AEFAFNF NAPSRNQ NPRPLSI SHYVNSQ EGLHQRK HPQHAGG HTSHLSC WPSFPIP TPALQFP SFTWAPD APPYTAY SWPSHVP HPNLLSP NEYPPNP NPNWGPR TRRETPA HTTHHRN VETWRPN ERFIDTQ TTHQFPF 23 Phage display (common panning) 3 PMID:17907769 Melanin Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Melanin was purified from MNT-1 human melanoma cells. The final eluate yielded approximately 50 individual clones; 24 clones from the plate were chosen at random and sequenced. Not 24, but 23 unique sequences were given in the published paper. Among them, 8 best tumor melanin-binding phages were identified by ELISA. Of the 8 phages, 3 best binding phages were identified as HTTHHRN, NPNWGPR and TTHQFPF. 370 HPMHFPS(11) YPRWQIP(6) SVFWMIP(6) YPQLPFT(4) HPMHFPC(1) HHYAFSV(1) SPNVNAN(1) 7 Phage display (common panning) 3 PMID:17711505 HBJ127 Natural killer cells antigen CD94 Not determined. Not determined. Ph.D.-7 phage display library (X7) Thirty clone were randomly picked and sequenced after the third round of panning. Seven unique sequences were obtained. 371 PSRALL(1) GRRALC(1) HRALSR(1) HRALRG(1) RAL 4 Phage display (common panning) 3-4 PMID:17252013 Kasumi-1 acute myeloid leukemia cells Not determined. Not determined. Not determined. X6 phage display library After the third and fouth rounds of selection, 11 and 12 clones were randomly picked and sequenced respectively. PSRALL and GRRALC were from the 3rd round; HRALSR and HRALRG, the 4th round. 372 CPLDIDFYC(7) CRGLGRALC(1) 2 Phage display (common panning) 3-4 PMID:17252013 Kasumi-1 acute myeloid leukemia cells Not determined. Not determined. Not determined. CX7C phage display library The template of CPLDIDFYC might be Vascular Cell Adhesion Molecule 1 (VCAM-1), since the α4β1 integrin (VLA-4) has been identified as the target of CPLDIDFYC. After the third and fouth rounds of selection, 11 and 19 clones were randomly picked and sequenced respectively. CRGLGRALC was from the 3rd round; CPLDIDFYC, the 4th round. 373 AWYPLPP(37) VQKHVHP(2) LGVCPPS(1) HAIYPRH(1) TQAKPKL(1) AATNLGI(1) QLHRHHH(1) APNLRPQ(1) KPVQLDH(1) KMLQSSV(1) WSYHRLP(1) 11 Phage display (subtractive panning) 3 PMID:16806673 Hepatocellular carcinoma cell line HCCLM3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) HCCLM3 has a high metastatic potential. The phage-displayed random peptide library was first screened with MHCC97L which is an HCC cell line with a low metastatic potential. The unbound phages remaining in supernatant were then screened HCCLM3. AWYPLPP was identified as a specific peptide ligand that binds to the cell surface of highly metastatic human hepatocellular carcinoma (HCC). This peptide was able to promote in vitro invasion of highly metastatic HCC cells by activating matrix metalloproteinase-9 and in vivo lung metastasis of HCC tumors. 374 GSSSWWQRWWPPW(14) GPMESLQAFWPPW(1) GVFLLKQVPQPSH(1) GRLWWLQLFEPGH(1) GLRKVPQSVPPDM(1) GHFLKPQVLRPTR(1) GQFMMRQYWPPVH(1) GLLKYQQWASPLC(1) GYFWYDQPWQPEQ(1) GRNHYIQRDNPVS(1) GVFHVLQNAIPQY(1) GTMPNMQHHDPAR(1) GTRYLVQYLFPHL(1) GRPATQQGLTPAR(1) GYIGTYQQWNPPP(1) 14 Phage display (common panning) 4 PMID:17268054 Human epidermoid carcinoma A431 cells Not determined. Not determined. Not determined. Tat-based pCANTAB-5E library library Cells on culture plates were incubated with the peptide-phage library for 2h at 37°C with shaking every 15min during the round of panning. Following this, the cells were washed 20 times with PBS at room temperature. After washing, the cells were lysed. 100ml lysate was used to infect E. coli and then used in the next round of selection. The GSSSWWQRWWPPW peptide did not exhibit cytotoxicity when recombined with PSIF. This result indicated that this tryptophan-rich peptide binds to the cell membrane but does not penetrate through to the cytoplasm. 375 IQSPHFF(19) HHSHRHH(4) HHHHLSM(2) VHPPLLT(1) KNVHVPL(1) GGHSTPE(1) DVTTPSP(1) 7 Phage display (subtractive panning) 4 PMID:17653285 Genhance 680, GH680 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) GH680 was covalently coupled to BSA and then immobilized on Nunc Maxisorp plates for selection. A subtraction well containing a BSA-only sample was prepared. The subtraction step was designed to filter BSA-and/or plastic-binding clones. IQSPHFF has nanomolar affinity for near infrare fluorochromes. The developed peptide sequence allows detection of NIR dyes in a wide range of assays including ELISA, flow cytometry, high throughput screens, microscopy, and optical in vivo imaging. 376 CSNRDARRC CANKDVRRC CANLDTRRC CPNQDSRRC CPNGDERNC CVNNDGRLC C-x-N-x-D-x-R(2)-C 6 Phage display (subtractive panning) 2-4 PMID:17259343 HT-1376 human bladder tumor cell line Not determined. Not determined. Not determined. CX7C T7 phage display library The tumor cells were isolated from tumor xenografts in nude mice prepared using the HT-1376 human bladder tumor cell line. Phage that bound to normal bladder cells isolated from mouse bladder tissues, normal rat kidney cells, or human umbilical vascular endothelial cells were subtracted during the biopanning procedure. After the second round of screening, the numbers of phage were enriched approximately by 1,000-fold and did not increase in further steps. Ninety-six phage clones were picked from the enriched phage library after the second and third rounds of screening. Twenty of the 96 clones showed selective binding to HT-1376 cells. These clones were sequenced. Six sequences were given in the published paper. It has been shown that CSNRDARRC selectively targeted and bound to primary bladder tumor tissues and can be used to detect tumor cells in urine. 377 AFNMFDFGV VTKSLRVFG SESHRVKSA QLGPPV 4 Phage display (common panning) 1-2 PMID:17129616 Anti-HAV polyclonal antibody IgG Hepatitis A virus, HAV Not determined. Not determined. J404 phage display library (X9) Mouse anti-human IgG coupled magnetic microbeads were blocked overnight at 4 ◦C in PBS/0.1% BSA (w/v). These beads were then incubated overnight at 4 ◦C with anti-HAV serum and washed four times with PBS/0.1% BSA (w/v)/0.1% Tween 20 (v/v) (PBSB-T). The beads were further blocked with excess of UV-killed wild-type phage M13K07 particles for 4 h at 4 ◦C and then used to screen the library. Since there was no enrichment after a second round of affinity selection, the 75 selected clones from the first round of panning, were tested. After a series of immunoscreenings, 4 clones are found to be disease-specific. Among them, VTKSLRVFG was recognised by 92% of the positive sera. Three of the four cloned peptides showed similarity in their amino acid sequences with at least one of the VP3 and VP1 antigenic proteins of HAV. 378 SVSVGMKPSPRP(25) WPLHTSVYPPSP(11) NTLPPFSPPSPP(5) SFPDSNIAPSSP(3) QHAPSNSKSVLT(2) WPTYLNPSSLKA(1) GPSGNLHIRPAS(1) SPLLSTRAVQLS(1) SPMFTMIQGDAQ(1) VNSHQALWSPAQ(1) STLPPPLRFANV(1) SFNQPYLYKTAF(1) YHTRIALPDNLP(1) AQSTAFQKPLLM(1) 14 Phage display (in vivo) 5 PMID:18006841 Non-small cell lung cancer (CL1-5) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) CL1-5 cells (lung cancer cell line) were injected s.c. into SCID mice to produce lung cancer xenografts. The paper suggested that PSP motif was crucial for peptide binding to the tumor neovasculature. The most frequent peptide SVSVGMKPSPRP was shown to be a good candidate for targeted drug delivery to solid tumors. 379 PPWDFDAGEGIH(1) DYAWDDFYAMGD(1) 2 Phage display (common panning) 3 PMID:17371999 Seam 3 monoclonal antibody Neisseria meningitidis serogroup B capsular polysaccharide (MenB CP) Not determined. Not determined. X9 phage display library 380 DYAWDQTHQ(3) DAGEGGPRV(2) DAGDSGYLT(1) DAGDHSHPQ(1) DAGEVYPGP(1) DAGDSAYSQ(1) DAGDHRAAA(1) EFDAGDVLL(1) D-A-G-[ED] 8 Phage display (common panning) 3 PMID:17371999 Seam 3 monoclonal antibody Neisseria meningitidis serogroup B capsular polysaccharide (MenB CP) Not determined. Not determined. X12 phage display library DYAWDQTHQ has been shown to be a promising lead candidate for the development of an effective and affordable anti-MenB vaccine. 381 ISYEHPLLTGLLLEQRHVD LDGDVSKLHPLLVNFLKGD VESSHPLLVGLLGKQQATV SEGLHPLLTALLAPQVTQW ALHDQQSLHPLLAEALLRD PMVQHPWLVGLLVSAEGVS DLAEPVKLHPLLSMYLSNL VAQMHPGLSELLLASHFHQ VHVTMPFLTSLLTGQPTSV MPLHSPYLLSLLTGSGQSL VDLEYPMLVHLLQYGRELS PVETHPRLLKLLTRAEYPM TEVTHPLLASLLQYGVQVS ATTPPTQLHPLLTQFLRTD SSLEHPWLTMLLGKPHNMW PVTAYPMLAELLLPTGQLR MSQSVSDLHPLLGLLLTAD VATNHPWLTMLLNGQASGE PLALHPLLASLLDPGRQLV TQMEYPLLTQLLGGRTMEE LLEAHPLLTGLLGASSLEL ILEEHPLLASLLTETSRGM LSQEYPWLTRLLDSGQLGS TWLEYPLLTDLLLPSWRQD SAADHPSLLRLLTGDFPYE ISYEHPLLTGLLLDSVMW LELEATSLHPLLLQLIKQV PVFEHPRLVGLLVGTDIAS LGESHPLLMQLLTENVGTH VEENPLLLEMLLSASKASS MVAQNPLLGGLLLGSMDTM PTSSPLLLDLLLGRPVETV PFYTHPMLSSLLLGPPSMT PTVEHPMLSSLLRGAANEF TVVQHPWLTGLLIGSGQDA SLEAHPLLSALLQDQMGGD PLSDHPMLLELLNHNSMRN HQEHPLLLHMLLGGSQEVE NHETHPLLSKLLLSPLEQT MSDHPLLLSYLLETTAVLP GVLDHPLLFQLLSSETRVL IAGEHPMLWGLLRAGGLPE PADNMPLLRTLLLGSGIEV TIDSHPMLHSLLSQRGLVA VAHTHPLLVGLLKGSITGP ALSDSPLLLSLLLGHPDWQ TYVDHPLLAELLLGEVGMN ADGSPMELHPLLSRLITAP PVEGHPWLLGLLVGAEVDV ALSSPSDLNPLLTSLLSQS MTLEHPGLTALLASQSLMG SVDQHPMLMDLLTGLKRAE MVEHPRLLSLLLDRSDVPM H-P-L(2)-x(2)-L(2) 53 Phage display (common panning) 3 PMID:17595321 Peroxisome proliferator-activated receptor gamma, PPAR-gamma PPAR-gamma coactivators Not determined. Not determined. X7-LXXLL-X7 M13 phage display library PPAR-gamma was expressed in Sf9 insect cells and biotinylated. 382 SLLHPLLVDLLQMGGAS SSVLNPLLAELLASGSS SYLLDLLLGTSDVTTNV L-x(2)-L(2) 3 Phage display (common panning) 3 PMID:17595321 Peroxisome proliferator-activated receptor gamma, PPAR-gamma PPAR-gamma coactivators Not determined. Not determined. SS-X16-S M13 phage display library PPAR-gamma was expressed in Sf9 insect cells and biotinylated. Of the 130 peptides sequenced, 53 exhibited unique sequences. 383 AVGLSPDGSRGV(10) GGSGDSRPPILG(1) TDTRPAHTGDAQ(1) PHIQADTRPHA(1) PDRPVPSLPITW(1) TPDSRGIHGAPS(1) PDGRPSPGHLPA(1) LLADTTHHRPWT(1) P-D-[TSG]-R-P 8 Phage display (common panning) 4-5 PMID:17725384 Anti-MUC1 monoclonal antibody PR81 Mucin-1, MUC-1 Not determined. Not determined. Ph.D.-12 phage display library (X12) The library was first screened against an irrelevant monoclonal anti-digoxin antibody that would not be expected to bind specifically to MUC1-specific epitopes. Additionally, in order to omit those phages that bind to BSA, the library was screened against BSA-coated wells. Fourty clones were randomly selected from each of the fourth and fifth rounds of biopanning. Among the 80 clones, 17 phage clones that showed higher binding activities (7 clones from the fourth round and 10 clones from the fifth round) were sequenced. 384 GHLSDWVYVPMR(5) VSMDDGWVFVQP(4) SHKSDWIFLPNA(3) AHKSDNWVFLPE(1) THINEKWVFLPQ(1) 5 Phage display (common panning) 4 PMID:17916189 Gamma-aminobutyric acid receptor-associated protein Calreticulin 3DOW, Not determined. Ph.D.-12 phage display library (X12) In fact, recombinant GST-GABARAP fusion protein was used for screening. Seventy randomly chosen true positive clones were sequenced. Only those with high occurrence and/or high affinity to GABARAP were given in the original paper. Calreticulin and GABARAP interact with a dissociation constant Kd=64 nm and a mean lifetime of the complex of 20 min. Thus, the interaction between GABARAP and calreticulin is the strongest so far reported for each protein. 385 KSLSRHDIHHHH(4) 1 Phage display (common panning) 4 PMID:17214965 Anti-HGF-cMET monoclonal antibody SFN68 Hepatocyte growth factor-hepatocyte growth factor receptor complex (HGF-cMET) Not determined. Not determined. Ph.D.-12 phage display library (X12) Ph.D.-7, C7C and 12 libraries were used in panning. Of 14 clones with specific reactivity, four shared the amino acid sequence KSLSRHDHIHHH. This is the only mimotope given in the original paper. 386 RVPPRYHAKISPMVN(20) 1 Phage display (common panning) 4 PMID:17005302 Anti-GMDP monoclonal antibody E6/1.2 N-acetylglucosaminyl-beta1-4-N-acetylmuramyl-alanyl-d-isoglutamine (GMDP) Not determined. Not determined. f3-15mer phage display library (X15) Twenty clones were selected and sequenced. All of them has the identical sequence RVPPRYHAKISPMVN. 387 YQDSAKT(17) NDRGLLA(5) VSVGMLW(4) QDTGLLA(2) SNPGWLI(2) 5 Phage display (common panning) 4 PMID:17761669 Beta amyloid peptide (12-28) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Only peptides found in two or more clones are shown. 388 SVLDRQR(15) SARTFLP(3) VDANKFL(2) SHREPLQ(2) 4 Phage display (common panning) 4 PMID:17761669 Beta amyloid peptide (25-35) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Only peptides found in two or more clones are shown. SVLDRQR was found to be effective blocker of the toxic effect of β-amyloid on cultured neuronal cells. 389 LGSYKPS(7) IQLHPRL(4) YGAAKSG(3) QATGLLA(3) SFHPPSM(2) QDTGLLA(1) EWGGAQF(1) 7 Phage display (common panning) 4 PMID:17761669 Beta-amyloid protein 42 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) LGSYKPS was found to be effective blocker of the toxic effect of β-amyloid on cultured neuronal cells. 390 NIPTPKP(3) SARTFLP(2) QATGLLA(2) IQSPHFF(2) SMPTYNK(2) GTHVQAT(2) VSVGMLW(2) NDRGLLA(1) 8 Phage display (common panning) 4 PMID:17761669 Beta amyloid peptide (1-20) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) VSVGMLW was found to be effective blocker of the toxic effect of β-amyloid on cultured neuronal cells. 391 CPYANHLLC(11) CPYANPSMC(4) CPYHNQFLC(2) CPKSAHKHC(2) CLYTNPALC(1) P-Y-x-N-x(2)-L 5 Phage display (subtractive panning) 3 PMID:18025245 Anti-PYANPSL polyclonal antibody IgG Peptide PYANPSL Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 392 CWAANPSMC(6) CRALNPSMC(5) CPINPSMAC(4) CPFNPSMAC(3) CLNPSSSQC(2) CPYNPSAHC(1) CYNPSSSWC(1) NPSM 7 Phage display (subtractive panning) 3 PMID:18025245 Anti-WAANPSM polyclonal antibody IgG Peptide WAANPSM Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 393 NPGTCKDKWIECLLNG(9) LKNYCRKCSNRCTPTG(4) NLIWCRKEFARCTSDM(3) 3 Phage display (subtractive panning) 3 PMID:17803452 Tumor-associated glycoprotein (TAG-72) Not determined. Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) Twenty-three clones were selected for sequencing. However, only above squences were given in the original paper. NPGTCKDKWIECLLNG showed evidence of significant specific binding when presented as the radiolabeled phage to tumor in vivo and especially in vitro with cells and solid tumor. 394 EDIKPKTSLAFR(6) TQPADLQTHNHN(1) FDHSSKWTRTSP(1) YSHNTITNLYFS(1) WPRYAESTLQLR(1) 5 Phage display (in vivo) 3 PMID:17687498 Nasopharyngeal carcinoma cell CEN-1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After the fifth round, all sequenced clones have the same sequence EDIKPKTSLAFR, which might be a candidate as diagnostics and therapeutics for nasopharyngeal carcinoma. 395 SVSVGMKPSPRP(2) IHPPTPPLTMLR(1) HLWPVNVASAQS(1) GSSRAQMTSHRP(1) SVGSSPHPGRHT(1) SDSPHGAVPPLA(1) HWTPWQHVSSFW(1) HPPDNARRIGCR(1) WSKNNKSPQLSP(1) GSMCPYVRWYTP(1) LNAAYSATFMAR(1) S-V-S-x(2)-M-x-P-S-P-R-P 11 Phage display (common panning) 6 PMID:18053412 l,2-Dioleoyl-sn-glycero-3-phosphatidylserine Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Ligands that bind to PS can be used for noninvasive imaging of therapy-induced cell death, particularly apoptosis. 396 CFHNHNPLC(20) CTVNTQSKC(10) CNHMPHPLC(5) CVGQKQESC(5) CLKAFHKEC(3) CHNQPQATC(3) CGPTPTKTC(2) CPLFSSKTC(2) CLTKSYTSC(2) CVTQHRGDC(2) CAPNSHWTC(1) CRLDHALQC(1) CILNNTPWC(1) CKDANSPHC(1) CPTRAALTC(1) CQKHNTRFC(1) CMPHTHRIC(1) CSQSHPRHC(1) CTQPRSTGC(1) CFPNTRATC(1) CSPVHQQSC(1) CPKSDHKFC(1) CAPKQTQHC(1) 23 Phage display (subtractive panning) 1 PMID:17324139 Mouse inflammatory small bowel Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) SQSHPRH has a specific high affinity for inflammatory bowel. 397 HPAIVHISPQWA(1) ISPAPHLLTSRF(1) HHVDSLPTLDWK(1) MPTSSTAPPPLI(1) QMVYGPLRSTEQ(1) HGTNQALSLLTP(1) KLPHQPPSAAVH(1) ANYFSSPIKHAT(1) NSLTSEPLRYGG(1) VLNPQTTVMPPL(1) SPWTSFLQWARG(1) HPPHNMHLPAFS(1) APFAHSGPLAFS(1) ATTSLTPTMANH(1) QFPPKLTNNSML(1) MPTLTRAPHTAC(1) TPLHPKSLMVWH(1) QATGPTTPTTSG(1) YTDNSLGTSVGK(1) LPRKTPDYLQTR(1) SNSMHLMTMTGL(1) TPSLEQRTVYAK(1) TSLRELPAEWSR(1) SVPPRYTLTLQW(1) LYSASTPPDPGG(1) SHGKPPSRSPWT(1) TLDWTKPPLRSG(1) FPMSSYKTYATP(1) YNLGQLEAQITS(1) ASQGYPEHRHAS(1) INTPANRNPVLG(1) ITLSATKGAAPS(1) HKSYLPVPSLYG(1) WDTNRNAASTPG(1) SPPPSNAGSHHV(1) QTTKLHIMDTGF(1) SYQTLKQHLPYG(1) THPPNPSVSIGG(1) S-P-T-P-S-[PL]-P(2)-S-A-G(2) 38 Phage display (subtractive panning) 3 PMID:17657770 Human blood outgrowth endothelial cells, HBOEC Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Before HBOEC screening, human umbilical vein endothelial cells (HUVEC) were used to remove non-specific binding phages. Fourty clones were sequenced. 38 different peptide sequences were deduced and 2 phage clones contained no inserts. 398 AMSFYNTPDAKE(1) TLCNNSMSPGCK(1) IEHTRYNSRTAY(1) QATKYHSANRHP(1) NYLHNHPYGTVG(1) GQSPHSYQPRTY(1) NNALLNRQLHFS(1) HQVPSHNNSLGP(1) 8 Phage display (common panning) 3 PMID:16958043 Beta-nerve growth factor, Beta-NGF Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) These peptide sequences were obtained when the pH of elution buffer was at 6.0. 399 RSTSPDPLNYVR(1) ALRTPLSHAPTK(1) NMTAVTKLPAPW(1) QMHPSIALPLWY(1) QMHPSIALPHWY(1) LDLALGQRPPRH(1) DARHLPPVTLST(1) SVSVGMKPSPRP(1) 8 Phage display (common panning) 3 PMID:16958043 Beta-nerve growth factor, Beta-NGF Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) These peptide sequences were obtained when the pH of elution buffer was at 4.5. 400 SVSVGMKPSPRP(2) DSAVDYTGWLRR(1) SVWYYXIHYDMX(1) SLTGAALGPSGW(1) VPHSKYPRGINA(1) QFSLPVAKLVNR(1) 6 Phage display (common panning) 3 PMID:16958043 Beta-nerve growth factor, Beta-NGF Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) These peptide sequences were obtained when the pH of elution buffer was at 2.8. 401 GQPTPRNAGLPL(3) SRLNVEPLTTYS(2) TTLHWASLTTGR(2) 3 Phage display (common panning) 4 PMID:17136792 Gonadotropin-releasing hormone promoter (GP) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) GP (from -813 to -1081) was immobilized on the surface of Streptavidin MagneSphere Paramagnetic Particles. The beads-DNA complex were then incubated with the library. Twenty clones were sequenced and 16 unique sequences were got. However, sequences appeared only once were not given in the original paper. 402 QSNPIHIITNTRNHP(3) KSNPIHIIQNRRNIP(2) KSNPIHIIKNRRNIP(2) SPKTTQPPNHIHSIP(2) 4 Phage display (common panning) 3 PMID:17577983 Human rotavirus Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Human rotavirus was cultured in MA104 cells. The purified rotavirus particles immobilized on solid phase were used for panning. These 4 peptides could specifically bind with rotavirus particles. The first 3 peptides can inhibit rotavirus infecting in vitro. QSNPIHIITNTRNHP showed the best efficiency-93% neutralization infectivity. 403 FQHPSFI(14) PLPTLPL(13) LPPQSFH(8) ATYQHAT(3) WAESKTF(2) HSALPKW(2) VSFPFGF(1) 7 Phage display (subtractive panning) 4 PMID:17622312 Hepatocellular carcinoma cell line HepG2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The normal liver cell line (L-02) was used firstly to carry out subtractive screening. FQHPSFI was shown to bind to the hepatocellular carcinoma cell lines (HepG2 and BEL-7402) and biopsy specimens, but not to normal hepatocytes, other different cancer cells, or nontumor liver tissues. 404 KTYQGPL(3) FSPEIRN(3) VHLGYAT(1) PHLNYSR(1) ANHTSPV(1) PHPLYQS(1) 6 Phage display (subtractive panning) 3 PMID:17332091 Colon cancer cell line SW480 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The phages bound to SW480 were treated with normal human intestinal epithelial cells for 2 rounds to eliminate the nonspecific phages. About 50 clone were sequenced. Only these sequences were given in the original paper. VHLGYAT was shown to be the most effective peptide in targeting SW480, HT29 cells and tumor tissues but not the normal human intestinal epithelial cells and control colon tissue. 405 QHKTSITGHHLEP TLPSPLALLTVH YLFSVHWPPLKA 3 Phage display (subtractive panning) 4 PMID:17947467 Hepatocyte growth factor receptor Hepatocyte growth factor 1SHY, 4K3J,4O3T,4O3U, Not determined. Ph.D.-12 phage display library (X12) S114 (NIH 3T3, transfected with the human genes for HGF/SF and Met) and SK-LMS-1/HGF (human leiomyosarcoma cell line autocrine for human Met and human HGF/SF) cells were used alternately to screen the library. NIH 3T3 cells were used for negative selection to remove nonspecific binders. Thus, the target molecule was thought to be Met. Since the three sequenced clones were able to compete with HGF in a dose-dependent manner, their template should be HGF. Thirty phage clones were picked randomly, all of which were found reactive against Met recombinant protein. They were then tested by competitive ELISA with or without HGF for Met binding. Three clones with the ability to compete with HGF in a dose-dependent manner for Met binding were sequenced. YLFSVHWPPLKA displayed the highest competitive ability (59.3% inhibition). 406 EDYELMDLLAYL(0.95) 1 Phage display (subtractive panning) 5 PMID:17504878 Thyroid follicular carcinoma cell line FRO82-2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The human embryonic kidney cell line 293 cells were used for a negative selection. Among 20 clones sequenced, 95% of all peptides showed the same sequence: EDYELMDLLAYL. 407 WRQTRKD(18) WHWRLPS(7) HAIYPRH(6) LRQTRKD(4) LPWHKHR(4) FHRHHNH(4) HFFHRHT(3) HWRHNHS(3) WPFHHHR(3) WHHKHRS(3) WPWHKHS(2) HKRPRNN(2) HFTTRQR(2) MPHIHRH(2) PRRESRF(2) KPAPRVH(2) KHTVTRR(2) HKTEHPA(1) HRSPHTP(1) SWRHHHH(1) SYSNWTP(1) QPPIYSA(1) QPPNYNA(1) KPRAGSF(1) KGPHTQV(1) IASGNSL(1) IPTLPSS(1) PTHRHRT(1) TAPGVST(1) LGPHTQV(1) 30 Phage display (common panning) 4 PMID:18221017 Arabidopsis Fip1 ortholog encoded by At5g58040 (AtFip1[V]) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Selection were carried out using the N-terminal 137 amino acids of AtFip1[V] as a target. A search of the Arabidopsis proteome using these Fip1-binding peptides as queries resulted in the identification of a number of putative Fip1-interacting proteins. One of these was the polyadenylation factor subunit, CstF77. This purported interaction was confirmed by yeast two-hybrid and in vitro assays. Mutation of the motif identified in the phage display screen eliminated the interaction, corroborating the results of the phage display screen. Eighty-two clones were sequenced. 408 WTITKHP(8) WRQTRKD(5) HAHTIST(5) KPVQLDH(3) HKRPRNN(2) ERYMGLP(2) IQLHPQH(1) SLDALLS(1) HAIYPRH(1) LVGNYTP(1) SILPYPY(1) TAPHPVL(1) HWRHNHS(1) 13 Phage display (common panning) 4 PMID:18221017 Arabidopsis Fip1 ortholog encoded by At5g58040 (AtFip1[V]) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Selection were carried out using the N-terminal 137 amino acids of AtFip1[V] as a target. A search of the Arabidopsis proteome using these Fip1-binding peptides as queries resulted in the identification of a number of putative Fip1-interacting proteins. One of these was the polyadenylation factor subunit, CstF77. This purported interaction was confirmed by yeast two-hybrid and in vitro assays. Mutation of the motif identified in the phage display screen eliminated the interaction, corroborating the results of the phage display screen. Thirty-two clones were sequenced. 409 WSYSWDGFW(6) SHLTWDGTW(6) YTVAWDGTW(3) LTTEWSGTW(2) GRLQWDGAW(2) KSYEWPGTW(2) MRTEWTGVW(2) RSIEWSGLW(1) KSASWDGLW(1) VVIQWDGAW(1) RTVEWSGNW(1) AKTVWDGRW(1) KSLLTTMAS(1) GWGMEALGA(1) RSPN(1) W-x-G-x-W 15 Phage display (subtractive panning) 6 PMID:18200546 Glioma cell line 9L Not determined. Not determined. Not determined. LL9 phage display library (X9) Rat plasma was used as adsorbent before the first round. Before the the second and each subsequent round of panning, phages were preadsorbed on rat blood cells. After the fourth and fifth panning rounds, 15 randomly selected phage clones were propagated and sequenced. RSIEWSGLW-coupled liposomal nanocarriers enhanced drug uptake by 9L cells by 500% compared with conventional liposomal nanocarriers, and significantly increased cytotoxicity. 410 CRGDNFQC(9) CDVFTYWMRRDC(9) CDVFEVWMGRVC(4) CRGTQNTMRARC(1) D-V-F-x(2)-W-M-x-R-x 4 Phage display (subtractive panning) 6 PMID:18200546 Glioma cell line 9L Not determined. Not determined. Not determined. CL6 phage display library (CX6C) Rat plasma was used as adsorbent before the first round. Before the the second and each subsequent round of panning, phages were preadsorbed on rat blood cells. After the fourth and fifth panning rounds, 15 randomly selected phage clones were propagated and sequenced. 411 QMAFYSHHRSSP VPQGSFYSSWYT QSLPQMRFYDS 3 Phage display (common panning) 3 PMID:18700874 Anti-ACE monoclonal antibody CG1 Angiotensin-converting enzyme, ACE Not determined. Not determined. Ph.D.-12 phage display library (X12) CG1 was selected from hybridomas, which were developed from mice immunized with soluble recombinant human tACE purified from culture medium of CHO cells. Its isotype: IgG1, Seven phage clones were isolated and sequenced. However, only the above three squences were given. Their occurrences were not clear. 412 LPQQLGKLLACC WCCAGKHTVTHS FGCLGKLVCDPPY MCQGKNICTTIQ LPCHSKYPCIAS IGMCKMKAPCAT YQCERKAPCSTY LLACTMKLPCSV ALQCQYKVPCLV HLSICDSKLICH ISMCKEKEVCQT STCSAKGMCTTW TPCTYKMTCTTK C-x(2)-K-x(2)-C 13 Phage display (common panning) 3 PMID:17046088 Anti-B-subtype HIV-1 polyclonal antibody pre-HAART IgG from Patient 1 Envelope glycoprotein gp160 Not determined. Not determined. Ph.D.-12 phage display library (X12) The template should be gp41 since the mimotopes are similar to CSGKLIC of gp41. Among 33 sequenced clones, 26 clones have CXXKXXC motif with 13 unique sequences. Seven sequences without CXXKXXC motif were not shown in the original articles. 413 HACTGKLRCTTT THQCLGKLQCGV LPQQLGKLLACC WCCAGKHTVTHS HLSICDSKLICH ACCTFKNVTTKP SMCSLKTACTTA C-x(2)-K-x(2)-C 7 Phage display (common panning) 3 PMID:17046088 Anti-B-subtype HIV-1 polyclonal antibody post-HAART IgG from Patient 1 Envelope glycoprotein gp160 Not determined. Not determined. Ph.D.-12 phage display library (X12) The template should be gp41 since the mimotopes are similar to CSGKLIC of gp41. Among 30 sequenced clones, 13 clones have CXXKXXC motif with 7 unique sequences. Seventeen sequences without CXXKXXC motif were not shown in the original articles. 414 HACTGKLRCTTT LPQQLGKLLACC DVCMGKLVCTML LLACTMKLPCSV HLSICDSKLICH YGCHSKITCTTF TICSMKSACTTW SPNCEGKIICGS C-x(2)-K-x(2)-C 8 Phage display (common panning) 3 PMID:17046088 Anti-B-subtype HIV-1 polyclonal antibody pre-HAART IgG from Patient 3 Envelope glycoprotein gp160 Not determined. Not determined. Ph.D.-12 phage display library (X12) The template should be gp41 since the mimotopes are similar to CSGKLIC of gp41. Among 61 sequenced clones, 16 clones have CXXKXXC motif with 8 unique sequences. Forty-five sequences without CXXKXXC motif were not shown in the original articles. 415 CSDSWHYWC(10) CSDWQHPWC(8) CSDYNHHWC(4) CSDGQHYWC(2) CYDSWHYWC(1) CFDGNHIWC(1) CTDFPRSFC(1) CTQDRQHPC(1) CLSRYLDQC(1) C-S-D-x(2)-H-x-W-C 9 Phage display (subtractive panning) 3 PMID:17517791 Vascular endothelial growth factor receptor 3, VEGFR-3 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Human IgG1 Fc was used firstly to remove Fc-binding phages. The phage displaying peptide CSDSWHYWC exhibited the highest affinity to VEGFR-3 in phage ELISA and the chemically synthesized CSDSWHYWC could bind to VEGFR-3 specifically in a dose-dependent manner. In addition, the flow cytometry assay and immunoflourescence showed that the FITC labelled CSDSWHYWC could bind to VEGFR-3 positive carcinoma cells with specificity. 416 CIGSPSTNC(14) CSFHYQNRC(3) CRQSRKRPC(1) CSRTSSRTC(1) CELSPSSRC(1) CTRWAGRPC(1) CLKRSKLRC(1) 7 Phage display (common panning) 5 PMID:17097662 Anti-mycobacterial HSP60 polyclonal antibody Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 417 CNPNNLSHC(1) CKNFTHTDC(1) CTTASGARC(1) CTDLLPRHC(1) CPTAPLHMC(1) 5 Phage display (common panning) 4 PMID:17482566 Paracetamol(4-acetamidophenol, 4AAP) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The identified NPNNLSH-phage clone displayed functional binding properties against paracetamol in solution, able in a peptide sequence-dependant manner to prevent the in vitro hepatotoxicity of paracetamol and reduce (~20%) the permeability of paracetamol across a semi-permeable membrane. 418 CNPNNLSHC(1) CRAPSQTVC(1) CDGNSRTQC(1) CTTLTKTFC(1) CTLRSATAC(1) CSHLHSPLC(1) CENTQKNSC(1) CSQGRLGQC(1) CDRNGSNAC(1) CSQHSSRSC(1) CLNSHLQTC(1) CRTTSDALC(1) CTSDWRLHC(1) 13 Phage display (subtractive panning) 4 PMID:17482566 Paracetamol(4-acetamidophenol, 4AAP) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) After 4 rounds of 4-AAP selection, 2-AAP and 3-AAP were used sequentially to delete those phages also binding to the structural isomers 2-AAP and 3-AAP from the 4-AAP binding pool. After 4 rounds of 4-AAP selection, 2-AAP and 3-AAP were used sequentially to delete those phages also binding to the structural isomers 2-AAP and 3-AAP from the 4-AAP binding pool. The identified NPNNLSH-phage clone displayed functional binding properties against paracetamol in solution, able in a peptide sequence-dependant manner to prevent the in vitro hepatotoxicity of paracetamol and reduce (~20%) the permeability of paracetamol across a semi-permeable membrane. 419 CGPSEPIGAWDC CGPAEPRGAWVC CGPREPDGSWHC CGPVEPVGAWVC CGPWEPSGIWSC CGPVEPKGVWFC CGPYEPRGDWTC G-P-x-E-P-x(3)-W 7 Phage display (common panning) 3 PMID:18184085 Anti-CD4 monoclonal antibody 5145A T-cell surface glycoprotein CD4 Not determined. Not determined. CL10 phage display library (CX10C) The human monoclonal antibody 5145A against HIV-1 gp120 was isolated from an asymptomatic, seropositive hemophiliac. The epitope of this HuMAb overlaps the CD4-binding site of gp120. The antisera from mimotope fusion protein immunized rabbit did not compete with CD4 for binding to gp120, although 5145A is specific for the conserved CD4 binding site region of the HIV envelope protein gp120. Perhaps, the template is not CD4. GPYEPRGDWT and GPAEPRGAWV bearing phage clones had the highest apparent affinity, whereas GPSEPIGAWD-phage 2 had the lowest. Rabbits immunized with peptide-protein fusions produced antisera that bound to recombinant HIV-1 gp120, but did not bind to HIV-infected cells nor neutralize HIV. The antisera also did not compete with CD4 or antibodies to the CD4 binding site for binding to gp120. 420 YGLTALDPLFKP(4) AFAKPLASDPLF(2) KVLTAVDPLYYA(1) KPLAMDPLHVST(1) KPLASDPLGAMP(1) KLSATDPLAHYK(1) NLYAVDMLVLAR(1) VTADMASDMLHH(1) HIRLQAIDPMPP(1) HLGLMAMDHLPW(1) QTILAHDPVKMR(1) HPLASSPLFAAP(1) A-x-D-[PM]-L 12 Phage display (common panning) 3 PMID:17504878 Anti-A. paragallinarum strain 0083 polyclonal antibody Avibacterium paragallinarum strain 0083 (serovar A) Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage clones containing the peptide motif reacted with the target antibody and this interaction could be blocked, in a dose-dependent manner, by A. paragallinarum. Immunization with YGLLAVDPLFKP-expressing recombinant bacteria induced a specific serological response to serovar A. A. paragallinarum. The chickens given the recombinant E. coli showed significant protection against challenge with A. paragallinarum 0083. 421 CRDALGMIGQC(2) CRNRLGMIQQC(2) CRDGRGMMTQC(1) CRDSTGMMAQC(1) CRDSTGMVKQC(1) CRDSAGMVGMC(1) CRRRDGLIAQC(1) CRGLEGDIVYC(1) CRIRGGMGFAC(1) CVRMSDTMLYC(1) R-[DN]-x(2)-G-[ML]-[VI]-x-Q 10 Phage display (common panning) 3 PMID:18716770 Matrix metalloproteinase-9, MMP-9 Not determined. Not determined. Not determined. CL10 phage display library (CX10C) Recombinant human MMP-9 was immoblized through MMP-9 monoclonal antibody. 0.01% arabinose was used during phage production. 422 CRNRLGMIQQC(2) CRDSEGNVTSC(1) CRDSRGLVTQC(1) CRDSYGSVAQC(1) CRDNAGLVRQC(1) CRDKEGNIASC(1) CRNKAGLVTQC(1) CMRNGQVFMVC(1) CEPHSMRPC(1) R-[DN]-x(2)-G-[ML]-[VI]-x-Q 9 Phage display (common panning) 4 PMID:18716770 Matrix metalloproteinase-9, MMP-9 Not determined. Not determined. Not determined. CL10 phage display library (CX10C) Recombinant human MMP-9 was immoblized through MMP-9 monoclonal antibody. 0.01% arabinose was used during phage production. 423 CRNDSGLVVQC(1) CRNQFGTRVIC(1) CRQRNGSVIQC(1) CRTTTGEVAQC(1) CMRLPSGFIHC(1) CNYALKHKWFC(1) CRDSTGMVKQC(1) R-[DN]-x(2)-G-[ML]-[VI]-x-Q 7 Phage display (common panning) 3 PMID:18716770 Matrix metalloproteinase-9, MMP-9 Not determined. Not determined. Not determined. CL10 phage display library (CX10C) Recombinant human MMP-9 was immoblized through MMP-9 monoclonal antibody. 0.001% arabinose was used during phage production. 424 CRDREGNVMRC(2) CDVSQWGEALC(1) CDVEQWGKAVC(1) CRDASGMVDLC(1) CRNELGMVAQC(1) CRDREGLVRQC(1) CRSREGDVMKC(1) CRNGVGLVIQC(1) R-[DN]-x(2)-G-[ML]-[VI]-x-Q 8 Phage display (common panning) 4 PMID:18716770 Matrix metalloproteinase-9, MMP-9 Not determined. Not determined. Not determined. CL10 phage display library (CX10C) Recombinant human MMP-9 was immoblized through MMP-9 monoclonal antibody. 0.001% arabinose was used during phage production. 425 HCWHYKFC(16) 1 Phage display (common panning) 6 PMID:18591232 serine repeat antigen 5, SERA5 Not determined. Not determined. Not determined. X8 and CX8C phage display library pool Recombinant SERA5 enzyme domain was used for panning. Following six rounds of panning, 48 individual clones from the pool were propagated and tested for the ability to bind to SERA5. All of the clones from the 8-residue library were positive and displayed essentially no binding to the blocking protein. From these positive clones, 16 from the library pool were selected and sequenced. 426 SRAVCPVEVCRWVV(3) FRAVCPPAVCYWHS(3) EGWCEMHSRWCVVS(3) SVYCEKFRWVCEFR(3) LVCHPAVPALLCAR(1) RCCAPYVPAFFCEC(1) 6 Phage display (common panning) 6 PMID:18591232 serine repeat antigen 5, SERA5 Not determined. Not determined. Not determined. X14 and CX14C phage display library pool Recombinant SERA5 enzyme domain was used for panning. Following six rounds of panning, 48 individual clones from the pool were propagated and tested for the ability to bind to SERA5. Only approximately one-third of the clones from the 14-residue library pool were positive and displayed essentially no binding to the blocking protein. From these positive clones, 16 from the library pool were selected and sequenced. Addition of LVCHPAVPALLCAR to parasite cultures compromised development of late-stage parasites. 427 CVSEDIYDAC(22) CEFQQWSGKC(8) CNHVCSRLGC(7) CNETTVREYC(4) CIEETARKGC(2) CNELHMKQHC(2) CNNATFEDGC(1) CNNATVEDEC(1) CEFLQWSGKC(1) CETGERIVLC(1) CDEKRGPNEC(1) CHSWKPDKLC(1) 12 Phage display (common panning) 5 PMID:18616802 Acute myeloid leukemia cells Not determined. Not determined. Not determined. Display PHAGE system library (CX6C) The peptides bound to leukemia cells, were internalized and could induce proliferation and/or differentiation in the target patient cells. 428 CTNPESLTC CKHHNWWTC CSLRTYAAC CFSSARLSC CQRTTEANC CTGYHNNTC CSQWLQGS CNSMWVRNC CHTLPHTKC CDLLLPGRC CSTTNATWC CHGSTKWAC CYRTQFTQC CTPLSTLQC CTPGRSATC CNPMHSRTC CKPSTSGQC CNKQFSAAC CSPYAKHNC CHSLRNAFC CQGSPYRHC CSAGAPEFC CTQSGLLSC CQVLNGNHC CKSFTTTRC CTYPYPKFC CKSTFSPNC CTSAAVHMC CQPHLPWHC CQTTNWNTC CSASTESLC CLVRNLAWC 32 Phage display (common panning) 6 PMID:18391034 Sin Nombre virus (SNV) strain SN77734 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) SNV was propagated and the titer was determined in Vero E6 cells. The preparation of UV-inactivated SNV was used for panning. The phages were eluted with anti-SNV IgG antibodies from human convalescent plasma (HCP). In all cases, the isolated peptides were less effective at blocking infection than were the same peptides presented by phage. Sin Nombre virus infection involves the interaction between viral membrane surface glycoproteins and the human integrin 429 CTTHISQTC CHPSKALQC CERLNLPQC CTDNALKAC CYKMDNHTC CHVLDPHLC CSRNHILTC CVMSKHQHC CLMGSSHSC CPSHYTQAC CHADQLPMC CLPTPHHVC CQLSLAPYC CTFHSPRFC CYATTLGAC CGPSLRGVC CFNTHTANC CPGHHLSHC CNSPKGKPC CQWPGQSGC CNSSSPTAC CHQLMQNLC CQATTARNC 23 Phage display (common panning) 6 PMID:18391034 Sin Nombre virus (SNV) strain SN77734 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) SNV was propagated and the titer was determined in Vero E6 cells. The preparation of UV-inactivated SNV was used for panning. The phages were eluted with polyclonal anti-Gn-BSA IgG. They were obtained sera from a rabbit immunized with the bovine serum albumin (BSA)-conjugated peptide LKIESSCNFDLHVPATTTQKYNQVDWTKKSS, which corresponds to residues 58 to 88 of Gn from SNV strain SN77734. In all cases, the isolated peptides were less effective at blocking infection than were the same peptides presented by phage. Sin Nombre virus infection involves the interaction between viral membrane surface glycoproteins and the human integrin αvβ3. Some peptides selected are similar to human integrin beta-3 (CD61: P05106). 430 CLKDNHRSC(4) CTPRQSPIC(4) CYDPIWRTC(3) CCYTAALAC(3) CMLHAYAQC(2) CFLGFSQQC(2) CSVPINDSC(1) CDHRQGSSC(1) CAPYNTLAC(1) CSPHIIASC(1) CFSTNMKTC(1) CRTTGAQTC(1) CPLFKGMSC(1) CLPAYSTYC(1) CRDSSAHQC(1) CHANFLHMC(1) CSLNTRSQC(1) 17 Phage display (common panning) 4 PMID:18670594 Cysteine-rich protein 1, CRP-1 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) CRIP1 was digested with enterokinase to remove the His tag and then used as target. LKDNHRS and YDPIWRT have the highest affinities for CRIP1. With peptide structure information and the NMR structure of CRIP1, the higher-affinity LKDNHRS was computationally redesigned, yielding a novel peptide, LDGGGKG, whose affinity was predicted to be much improved. Synthesis of this peptide and saturation and competitive binding studies demonstrated approximately a 10-28-fold improvement in the affinity of LDGGGKG compared to that of either LKDNHRS or YDPIWRTA1 peptide. VRPMPLQ does have partial homology (6 of 7 amino acids, V/LRPMPLQ) to the laminin-G domain of contactin-associated protein 1 (Caspr-1), which is present in human intestine and might be involved in contactin-mediated cell signaling and in tumor metastasis and invasion. 431 VVFSTSV ISARSSP GDSMQQQ LQVHTQN WARSESP AALVRHT VRPMPLQ NQPPDSY APPWIAV LPSSYPP YTPLLPS 11 Phage display (subtractive panning) 3 PMID:18345013 Fresh human colonic adenomas Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Intact colon polyps freshly resected were used as the target. The binding of VRPMPLQ to HT-29 human adenocarcinoma derived cells was around 20-fold greater than its binding to Hs738.st/int nonmalignant human intestinal cells. The bindings of APPWIAV, LPSSYPP and YTPLLPS to HT-29 were similar to wild-type insertless M13 phages. The fluorescein-conjugated VRPMPLQ bound more strongly to dysplastic colonocytes than to adjacent normal cells with 81% sensitivity and 82% specificity. 432 NAVWLPQHPLRT SFHHLPIHPTAH NPISSFYLPQHP NSPWPLHPLRVF YFPYPQHPTTSN SLWLPWQPRHPP KLIIGSPYPMHP ANILAIHHPRHP SLNLHLPLHPTF VPHLPRHPLSSY NPFLPRHPSPML RLTLSPYPLHPL SFHHLPIHPTAD HPAHTMPRHPYT TNYPWLPKHPTS L(0,1)-P-[QR]-H-P 15 Phage display (common panning) 3 PMID:17847085 Anti-ErbB2 monoclonal antibody A21 Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) Fifteen phage clones which showed the strongest reactivity to mAb A21 (ELISA signals >1.0) and no crossreaction to another anti-ErbB2 mAb A18 were selected and sequenced. 433 KLLRIAP(4) ATWRIGP(4) KTLRIAP(1) KDVRIAP(1) KVVRIEP(1) NILSTLL(1) NVFNWKW(1) EFRWAWA(1) DWTWSWN(1) WSWGWMA(1) 10 Phage display (subtractive panning) 3 PMID:19081789 Anti-SHIV polyclonal antibody IgG Envelope glycoprotein gp160 Not determined. Not determined. Ph.D.-7 phage display library (X7) The rhesus monkey was infected with SHIV-1157ip, an R5 SHIV strain encoding env of a recently transmitted Zambian HIV-C. Its polyclonal IgG was immobilized by a rabbit anti-monkey IgG and used as target. Eluted phages were subjected to negative selection using pooled sera from non-infected control monkeys. 434 CSHGKLLAC(12) CTGKLQCIC(3) 2 Phage display (subtractive panning) 3 PMID:19081789 Anti-SHIV polyclonal antibody IgG Envelope glycoprotein gp160 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The rhesus monkey was infected with SHIV-1157ip, an R5 SHIV strain encoding env of a recently transmitted Zambian HIV-C. Its polyclonal IgG was immobilized by a rabbit anti-monkey IgG and used as target. Eluted phages were subjected to negative selection using pooled sera from non-infected control monkeys. 435 GPSKIFTWGWAF(12) IRPGPAAGGYPA(3) SLWTLHGSLISA(3) KMIHLGPQQTFP(2) SLLYSSEYSGIW(2) KLLSSNTYGIWM(2) IRPHPGHMYYSW(1) QVRMGPGQPDYL(1) VRLPPGASGYTP(1) KLLGYTTSAGIW(1) LCYHRDGSYPTS(1) SLLKHSLSAGIW(1) QHSWSCSGKLLC(1) SIWQTSGVLISY(1) 14 Phage display (subtractive panning) 3 PMID:19081789 Anti-SHIV polyclonal antibody IgG Envelope glycoprotein gp160 Not determined. Not determined. Ph.D.-12 phage display library (X12) The rhesus monkey was infected with SHIV-1157ip, an R5 SHIV strain encoding env of a recently transmitted Zambian HIV-C. Its polyclonal IgG was immobilized by a rabbit anti-monkey IgG and used as target. Eluted phages were subjected to negative selection using pooled sera from non-infected control monkeys. 436 WSRPRSL(16) 1 Phage display (common panning) 3-5 PMID:18309279 RNA-directed RNA polymerase NS5B Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Twenty-six clones were sequenced. The binding of WSRPRSL to HCV NS5B in vivo was shown by a yeast two hybrid assay and by subcellular colocalization analysis using immunofluorescence confocal microscopy. 437 TGPLPKE(8) 1 Phage display (common panning) 3-5 PMID:18309279 RNA-directed RNA polymerase NS5B Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Twenty-three clones were sequenced. 438 CATRLGRSSRC(18) 1 Phage display (common panning) 3-5 PMID:18309279 RNA-directed RNA polymerase NS5B Not determined. Not determined. Not determined. CX9C phage display library Twenty-six clones were sequenced. 439 CMAFFISRWRC(5) 1 Phage display (common panning) 3-5 PMID:18309279 RNA-directed RNA polymerase NS5B Not determined. Not determined. Not determined. CX9C phage display library Twenty-three clones were sequenced. 440 NYNLSRNLTWFY(5) 1 Phage display (common panning) 3-5 PMID:18309279 RNA-directed RNA polymerase NS5B Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Ten clones were sequenced. 441 CKPTGMPQC(1) CPTWALHLC(1) CPTWLSPAC(1) CRSPDMPFC(1) 4 Phage display (subtractive panning) 1 PMID:18569284 Colon cancer cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Laser capture microdissection (LCM) is performed to separate only cancer cells as the target used. Twenty clones were sequenced. Samples of tumor and noncancerous colon tissue were obtained at the time of colectomy from patients with operable colon cancer. Laser capture microdissection (LCM) is performed to separate only cancer cells as the target used. Initially, a subtraction step using non-cancerous colon tissue was performed. 442 CSPIQDRHC(2) CHMPTAQEC(1) CKQYASPWC(1) 3 Phage display (subtractive panning) 2 PMID:18569284 Colon cancer cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Laser capture microdissection (LCM) is performed to separate only cancer cells as the target used. Twenty clones were sequenced. Samples of tumor and noncancerous colon tissue were obtained at the time of colectomy from patients with operable colon cancer. Laser capture microdissection (LCM) is performed to separate only cancer cells as the target used. Initially, a subtraction step using non-cancerous colon tissue was performed. 443 CPTWLSPAC(2) CPFSPSLKC(2) CSPTKSNSC(2) CQTTRSPIC(1) CQHASPTNC(1) CQPTPRSTC(1) CPTPNHDHC(1) CPTSHRNSC(1) CPTTKLSTC(1) CSIHGPTRC(1) SPT 10 Phage display (subtractive panning) 3 PMID:18569284 Colon cancer cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Laser capture microdissection (LCM) is performed to separate only cancer cells as the target used. Thirty-seven clones were sequenced. Samples of tumor and noncancerous colon tissue were obtained at the time of colectomy from patients with operable colon cancer. Laser capture microdissection (LCM) is performed to separate only cancer cells as the target used. Initially, a subtraction step using non-cancerous colon tissue was performed. Phage clones displaying the SPT motif demonstrated 9-fold higher binding to colon cancer cells derived from a patient than insertless phage, while, recovery of the SPT phage from the colon cancer cell lines DLD-1 and HCT-15 was 7-fold higher than that of the control insertless phage. The binding of SPT phage to colon cancer cells from the patient was confirmed by immunofluorescence. Additionally, peptide SPTKSNS showed binding activity in the absence of mitogenic effects on colon cancer cells in vitro. 444 TTFYRRGA EASYRRKQ ASSYRTSR AAWYRTSR ARLYSRGA EAFYSQRF TRFYSRGR SFHYRMVG GTLFRSGN PNRWSTGA SSEWSMPY SSYISNFG LEARSAYH NAARSTGA EAKRSYHS EASRSATL 16 Phage display (subtractive panning) 6 PMID:18359858 Kallikrein-1 Not determined. Not determined. Not determined. fUSE55-based X8 phage display library From the third to the last round of screening, phage clones were randomly selected and plasmid DNA were isolated and sequenced to determine the displayed substrate peptide sequences. A total of six rounds of enrichment were performed for KLK1. 445 EAVRSAMW HLVRSWNG VGVRSVYG ASVRSAMY SKVRSAGA QMYRSSWG FGFRSVHG TAFRSAYG TAFRNSLG IGFRNAGA SLFRMVVL EAFRSSDQ ASSRSVKW AKSRSAGD AFLRMASL KVLRSATG YMTRSAMG ISTRSAIW WGWRYAET KEARSAYG 20 Phage display (subtractive panning) 5 PMID:18359858 Kallikrein-6 Not determined. Not determined. Not determined. fUSE55-based X8 phage display library From the third to the last round of screening, phage clones were randomly selected and plasmid DNA were isolated and sequenced to determine the displayed substrate peptide sequences. A total of five rounds of enrichment were performed for KLK6. 446 CPHMTAPFC CAPYSRFQC CQPPDRPMC CPDHERPMC CPLREHPMC RPMP 5 Phage display (subtractive panning) 3 PMID:18583245 Large intestinal cancer cell line LoVo Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Human normal large intestinal mucosal epithelial cells were used to remove non-specific phages to LoVo cells. 447 SPWSEPAYTLAP APWTEHSYYLSL VTHKTCPPACWP 3 Phage display (subtractive panning) 4 PMID:18782762 Claudin-4 Heat-labile enterotoxin B chain Not determined. Not determined. Ph.D.-12 phage display library (X12) CHO cells transfected with full-length Cldn-4 (CHO-Cldn-4) were used as the target. CHO control cells were used to remove phage specific for CHO determinants in the 2nd and the 4th round. 448 SFSIIHTPILPL(1) ELMNPLLPFIQP(1) HLPSTGNQYLSL(1) ETNWTHRPPLRV(1) EYRMAHLTPSLL(1) YHLQDSETLSLL(1) SPWYMTPSPNTA(1) SVSVGMKPSPRP(1) DPMTWTPSSVMR(1) TPHRLDWSPHLV(1) GSNPWNTWLTTL(1) NPFNQHLHAQHP(1) SESKDPTLWYPA(1) SFRLATPESSRV(1) SNNEPMLRYTGQ(1) 15 Phage display (common panning) 5 PMID:18347144 Hepatocellular carcinoma line Mahlavu Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Ninety-six phage clones were randomly isolated and used to react with hepatocellular carcinoma cells and normal epithelial cells by ELISA assay. Fifteen phage clones with higher hepatocellular carcinoma cell reactivity by ELISA and flow cytometry were selected and sequenced. In vivo, SFSIIHTPILPL phage homed specifically to tumor tissues but not to normal visceral organs in severe combined immunodeficient mice bearing human hepatocellular carcinoma xenografts. This homing ability could be competitively inhibited by synthetic peptide, SFSIIHTPILPL. Immunohistochemical staining confirmed that SFSIIHTPILPL phage localized to tumor tissues and that it could not be detected in SFSIIHTPILPL-competed tumor tissues. In addition, SFSIIHTPILPL phage recognized the tumor tissue but not nontumor tissue in surgical specimens from hepatocellular carcinoma patients, with a positive rate of 61.3% (19 of 31). With the conjugation of SFSIIHTPILPL and liposomal doxorubicin, the targeted drug delivery system enhanced the therapeutic efficacy against hepatocellular carcinoma xenografts through enhanced tumor apoptosis and decreased tumor angiogenesis. 449 QRFCTGHFGGLYPCNGP(23) GGGCVTGHFGGIYCNYQ(4) KIICSPGHFGGMYCQGK(3) PSYCIEGHIDGIYCFNA(3) NSFCRGRPGHEGGCYLF(1) G-H-F-G(2)-x-Y 5 Phage display (subtractive panning) 3 PMID:18272495 HEK293 cells transfected with hFcRn and hβ2m (293c11) Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) Not determined. 3M17,3M1B, TN phage display library pool In the motif, X is preferably a hydrophobic amino acid. The pooled phage libraries were screened with HEK293 cells transfected with hFcRn and hβ2m (293c11) by using competition with hIgG to select for phage capable of interfering with the IgG–FcRn interaction at pH 6. The subtraction step was performed twice by incubating the phage with the untransfected 293 cells. 450 CAYHRLRRC 1 Phage display (common panning) 2-4 PMID:18292083 Acute T-lymphoblastic leukemia Molt-4 cells Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Ninety-six clones were sequenced after the second, third, and fourth round of panning. There were 1, 17 and 55 clones respectively had the sequence CAYHRLRRC. CAYHRLRRC contains a lymph node-homing motif (CAY) and a cell-penetrating motif (RLRR). Binding of this ligand-directed phage to a large panel of leukemia/lymphoma cells and to patient-derived samples was much higher than to non-leukemia control cells. 451 CAPRIGLSC(1) CLGRPSTVC(1) CQKSPTNIC(1) CTKPIPHSC(1) CTSSAHRYC(1) CPSRSASHC(1) CLKSNLARC(1) CHETRSTIC(1) CEPTLYSMC(1) CPTKSHGTC(1) CKPSPTHGC(1) CDSRWQTLC(1) 12 Phage display (common panning) 4 PMID:18654983 Chymotrypsinogen A Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Heptamer sequences isolated from biopanning in buffer. 452 CPGSQRALC(2) CTDPAKKQC(1) CLTSGMTQC(1) CMTKLQKSC(1) CNSSHAKLC(1) CQQNNHRHC(1) CYSHGPQKC(1) CTTSLHPNC(1) CEHSTGRYC(1) 9 Phage display (common panning) 4 PMID:18654983 Chymotrypsinogen A Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Heptamer sequences isolated from biopanning in 50% acetonitrile (ACN). 453 CPWRAPGLC(1) CFPHETYQC(1) CHPFHPYVC(1) CNPHHLHSC(1) CTTVSHSTC(1) 5 Phage display (common panning) 3 PMID:18680214 Chitinase A Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) CPWRAPGLC was proved to be an inhibitor of chitinase A. 454 CRGLSHACG(3) CRHCDSPHG(2) CMRGIWTLC(1) 3 Phage display (common panning) 3 PMID:18680214 Chitinase B Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) CRHCDSPHG and CRGLSHACG were proved to be inhibitors of chitinase B. 455 CHETTLRRC(2) CELPALRLC(1) CLQDHSPFC(1) CDLYDSLSC(1) CMKSHRDKC(1) CAPFAHATC(1) 6 Phage display (subtractive panning) 3 PMID:17673295 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. Ph.D.-C7C phage display library (CX7C) In fact, the recombinant fusion protein HSA-FcγRIIa was used as the target. However, HSA-binding phages were removed using human serum albumin. For each round, panning parameters were wrote as (target) {wash} [elution]. Round 1, (100μg/ml HSA–FcγRIIa){nil}[0.1 mg/ml IgG]; round 2,(5μg/ml HSA–FcγRIIa){nil}[0.1 mg/ml IgG]; round 3,(5μg/ml HSA–FcγRIIa){nil}[0.1 mg/ml IgG]. 456 CWPGWDLNC(7) CSLKASFNC(1) CSWWTLSSC(1) CGTRPAPFC(1) C-W-P-G-W-x(2)-C 4 Phage display (subtractive panning) 3 PMID:17673295 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. Ph.D.-C7C phage display library (CX7C) In fact, the recombinant fusion protein HSA-FcγRIIa was used as the target. However, HSA-binding phages were removed using human serum albumin. For each round, panning parameters were wrote as (target) {wash} [elution]. Round 1, (100μg/ml HSA–FcγRIIa){0.1 mg/ml IgG}[10 mg/ml IgG]; round 2,(5μg/ml HSA–FcγRIIa){0.1 mg/ml IgG}[10 mg/ml IgG]; round 3,(5μg/ml HSA–FcγRIIa){0.1 mg/ml IgG}[10 mg/ml IgG]. The strongest binding clone CWPGWDLNC competed with IgG for binding to FcγRIIa and was inhibited from binding to FcγRIIa by the FcγRIIa-blocking antibody, suggesting that CWPGWDLNC and IgG share related binding sites on FcγRIIa. 457 CWPGWDLLC(6) CWPGWDLNC(1) CWPGWDEMC(1) CWPGWDMAC(1) CSERPSQQC(1) C-W-P-G-W-x(2)-C 5 Phage display (subtractive panning) 3 PMID:17673295 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. Ph.D.-C7C phage display library (CX7C) In fact, the recombinant fusion protein HSA-FcγRIIa was used as the target. However, HSA-binding phages were removed using human serum albumin. For each round, panning parameters were wrote as (target) {wash} [elution]. Round 1, (100μg/ml HSA–FcγRIIa){nil}[0.1 mg/ml IgG]; round 2,(5μg/ml HSA–FcγRIIa){0.1 mg/ml IgG}[1mM DTT]; round 3,(5μg/ml HSA–FcγRIIa){0.1 mg/ml IgG}[1mM DTT]. The strongest binding clone CWPGWDLNC competed with IgG for binding to FcγRIIa and was inhibited from binding to FcγRIIa by the FcγRIIa-blocking antibody, suggesting that CWPGWDLNC and IgG share related binding sites on FcγRIIa. 458 CTGQRTLYC(3) CWPGWDLNC(2) CPTALRIQC(2) CMPTTLGTC(1) C-W-P-G-W-x(2)-C 4 Phage display (subtractive panning) 3 PMID:17673295 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. Ph.D.-C7C phage display library (CX7C) In fact, the recombinant fusion protein HSA-FcγRIIa was used as the target. However, HSA-binding phages were removed using human serum albumin. For each round, panning parameters were wrote as (target) {wash} [elution]. Round 1, (100μg/ml HSA–FcγRIIa){nil}[0.1 mg/ml IgG]; round 2,(100μg/ml HSA–FcγRIIa){nil}[0.1 mg/ml IgG]; round 3,(100μg/ml HSA–FcγRIIa){nil}[0.1 mg/ml IgG]. The strongest binding clone CWPGWDLNC competed with IgG for binding to FcγRIIa and was inhibited from binding to FcγRIIa by the FcγRIIa-blocking antibody, suggesting that CWPGWDLNC and IgG share related binding sites on FcγRIIa. 459 CHIFQPLHC(1) CYDKQHSTC(1) CKLHLSKSC(1) CLPHSPRSC(1) CTSQKHLSC(1) CDLWFHPNC(1) CFTGYPPNC(1) CSQSQQPPC(1) CNALGMPIC(1) CNGPLFNIC(1) 10 Phage display (subtractive panning) 3 PMID:17673295 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. Ph.D.-C7C phage display library (CX7C) In fact, the recombinant fusion protein HSA-FcγRIIa was used as the target. However, HSA-binding phages were removed using human serum albumin. For each round, panning parameters were wrote as (target) {wash} [elution]. Round 1, (100μg/ml HSA–FcγRIIa){nil}[0.1 mg/ml IgG]; round 2,(100μg/ml HSA–FcγRIIa){0.1 mg/ml IgG}[1mM DTT]; round 3,(100μg/ml HSA–FcγRIIa){0.1 mg/ml IgG}[1mM DTT]. 460 CWPGWDLNC(4) CWPGWDLLC(1) CTGQRTLYC(1) CPTALRIQC(1) CQNSARQQC(1) CHAIWRSIC(1) C-W-P-G-W-x(2)-C 6 Phage display (subtractive panning) 3 PMID:17673295 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. Ph.D.-C7C phage display library (CX7C) In fact, the recombinant fusion protein HSA-FcγRIIa was used as the target. However, HSA-binding phages were removed using human serum albumin. For each round, panning parameters were wrote as (target) {wash} [elution]. Round 1, (100μg/ml HSA–FcγRIIa){0.1 mg/ml IgG}[10 mg/ml IgG]; round 2,(100μg/ml HSA–FcγRIIa){0.1 mg/ml IgG}[10 mg/ml IgG]; round 3,(100μg/ml HSA–FcγRIIa){0.1 mg/ml IgG}[10 mg/ml IgG]. The strongest binding clone CWPGWDLNC competed with IgG for binding to FcγRIIa and was inhibited from binding to FcγRIIa by the FcγRIIa-blocking antibody, suggesting that CWPGWDLNC and IgG share related binding sites on FcγRIIa. 461 CTLPHLKMC(10) CLYGNSDKC(3) CQHNHSKQC(3) CTKQHTSQC(3) CNPSLALHC(3) CSTGAHTQC(3) CDWTKPQSC(2) CTVKNGTLC(1) CQEGKMRLC(1) CSEHKTPMC(1) CQKQGHPSC(1) CTSTPFRIC(1) 12 Phage display (common panning) 4 PMID:18442789 Human B cell Burkitt lymphoma cell line Raji Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) MALDI-TOF mass spectrometry following immunoprecipitation assay showed that a potential template for the peptide CTLPHLKMC is a variable region of human immunoglobulin heavy chain which would be a specific phenotypic marker of Raji. The phage clone encoding CTLPHLKMC peptide sequence avidly bound to Raji cells compared with control phage clones. Furthermore, flow-cytometric analysis on the biotinylated synthetic CTLPHLKMC peptide demonstrated the high binding affinity to Raji cells in a dose-dependent manner whereas it has binding activity to neither human peripheral bloodmononuclear cells including normal B cell derived from healthy donors nor other leukemia cells including THP-1, HL-60, Jurkat and IM-9. 462 SWTLYTPSGQSK(4) HWYITTGPVREK(3) 2 Phage display (common panning) 3 PMID:18655820 Cadherin-2 Cadherin-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) In fact, the target used in the experiment is a chimeric protein composed of the N-cadherin ectodomain fused to the immunoglobulin G1 Fc fragment. The bound phages were eluted through adding 100 ml TBS containing 2 mM EDTA to each well and incubating for 10 min at room temperature with constant agitation. 463 NWFIDFPVYPPL(7) SWTLYTPSGQSK(6) AWQVHYSYVASS(2) KWELTYFANSFP(1) EWMIHYDSALTS(1) SWLAVWPATGAS(1) 6 Phage display (common panning) 3 PMID:18655820 Cadherin-2 Cadherin-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) In fact, the target used in the experiment is a chimeric protein composed of the N-cadherin ectodomain fused to the immunoglobulin G1 Fc fragment. The bound phages were eluted through adding 100 ml 0.2M glycine HCl to each well and incubating for 10 min at room temperature with constant agitation followed by neutralization with 1 M Tris-HCl, pH9.1 (20 ml/well). 464 CSERQALHGWC CDEKRALHNLC 2 Phage display (common panning) 3 PMID:18363340 Platelet glycoprotein Ib alpha chain von Willebrand factor (vWF) 1M10,1SQ0,1U0N,4C2A,4C2B, Not determined. CX9C M13 phage display library 465 CNERAALWNLC CTERWALHNLC CESRWWLRNAC 3 Phage display (common panning) 3 PMID:18363340 Platelet glycoprotein Ib alpha chain von Willebrand factor (vWF) 1M10,1SQ0,1U0N,4C2A,4C2B, Not determined. CX9W1-9C M13 phage display library CTERWALHNLC inhibited GPIbA-vWF-mediated platelet aggregation induced under high shear conditions. 466 HQPANDPSWYTG NTISGLRYAPHM 2 Phage display (common panning) 5 PMID:18067293 Tetragonal BaTiO3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Tetragonal BaTiO3 powder (99.9% purity) was used as the target. These peptides can induce the room-temperature formation of ferroelectric (tetragonal) BaTiO3 within 2 h from an aqueous precursor solution at near neutral pH. 467 CSKSSDYQC 1 Phage display (in vivo) 3 PMID:18440083 Rat small intestinal mucosal tissue Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The Ph.D.-C7C library was orally administered into overnight-starved, 12-week-old male Sprague-Dawley (SD) rats. After 1h retention, rats were sacrificed and phages from four representative organs (liver, lung, spleen, and kidney) were eluted and amplified for next round of panning. After the third round of biopanning, total 850 peptide sequences were determined from randomly selected individual phage recombinants from four representative organs, liver (from which 220 peptide sequences were identified), lung (218), spleen (204), and kidney (208). CSKSSDYQC was the peptide ligand which appeared 9 times (thrice in liver; twice in lung, spleen, and kidney) out of total 850 sequences selected by in vivo phage display. Phages displaying CSKSSDYQC showed a significantly higher affinity to small intestinal mucosal tissue compared with the native M13 phage. 468 VPGQKQHYVQPTAAN(23) 1 Phage display (subtractive panning) 3 PMID:18462752 Secretion chaperone Not determined. Not determined. Not determined. f88-15mer phage display library (X15) CsaA from Agrobacterium tumefaciens (AtCsaA) was biotinylated and used as the target. The initial library was incubated overnight with the streptavidin magnetic beads to remove streptavidin binding phage. Twenty-three clones were sequenced after the third round of selection. All clones had the same sequence VPGQKQHYVQPTAAN. The atomic coordinates of the AtCsaA structure with the bound peptide has been deposited in the PDB with the ID 2Q2H; the PDB ID for the AtCsaA structure without the peptide is 2Q2I. 469 CGIKVATQTQRC CRRKPEPLLPNC CPPVDRRHVAAC CMKRSPNEPHHC 4 Phage display (subtractive panning) 3 PMID:18462752 Secretion chaperone Not determined. Not determined. Not determined. LX10 phage display library (CX10C) CsaA from Agrobacterium tumefaciens (AtCsaA) was biotinylated and used as the target. The initial library was incubated overnight with the streptavidin magnetic beads to remove streptavidin binding phage. 470 AHVELMM(1) LTLAGAS(1) NVDGAVS(1) IAPTNGP(1) AENITTK(1) QQYDPMH(1) ALNMAPH(1) NTSPVPK(1) SFQLFHG(1) TWPYYPN(1) SAFDNPY(1) TRLNIPP(1) HSPPAMR(1) FNTLNSI(1) QLDTRLL(1) LPLNPLL(1) MLPHPTP(1) AETVKVV(1) STPRNSS(1) SPQQASA(1) VLTTRLA(1) WSPLHNT(1) ALLDPTV(1) YPGYFTK(1) YSKPSQM(1) YHYKTTS(1) HNFNVPF(1) SDNVKWN(1) AATPSEG(1) FTSSPSP(1) TSTVTHV(1) AADNTSG(1) ASSLRSV(1) SFKPAMH(1) NDPWQFH(1) TATDLSP(1) MYTSPLS(1) NMSGPLP(1) ADAITIG(1) HAYATFQ(1) QSPAAQP(1) KLPPSFP(1) MTSHTSG(1) LPKPWLN(1) GEVRTHA(1) GQFKLTK(1) AETVESC(1) STAGPVG(1) NAEGVRL(1) SIRLPSP(1) NNSMPGP(1) NTRLPVI(1) SKTDIPN(1) KSPPLMQ(1) AQVDVTV(1) HPSSYWT(1) FKMPLIS(1) LQTNWYS(1) RAPTPPF(1) 59 Phage display (in vivo) 1 PMID:18602466 Mouse digestive tract Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The Ph.D.-7 library was orally administered into 2 Swiss albino male mice weighing 25g. One hour later, phages passed through the intestinal barrier was evaluated by investigating its presence in the samples prepared from spleen. Statistical treatment of the obtained sequences did not support the notion that the GI translocation depends on the presence of any particular peptide sequence fused on the pIII coat proteins of the M13 phages. 471 APLLSSK(1) WHLPRPI(1) DVPAPKP(1) IRSPASY(1) DNRHSTP(1) SNYAVLL(1) HQNHNLS(1) AVYSPSR(1) TLPTIFH(1) AAGNRLP(1) SYTPYQP(1) NPPHLAS(1) FTSQAPT(1) DSLTARL(1) LPLHPVH(1) THLMSPI(1) STVSSMR(1) DHLVTAP(1) 18 Phage display (in vivo) 1 PMID:18602466 Mouse digestive tract Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The Ph.D.-7 library was orally administered into 2 Swiss albino male mice weighing 25g. One hour later, phages passed through the intestinal barrier was evaluated by investigating its presence in the samples prepared from blood. Statistical treatment of the obtained sequences did not support the notion that the GI translocation depends on the presence of any particular peptide sequence fused on the pIII coat proteins of the M13 phages. 472 CLEVSRKNC(9) CKRGGATAC(7) CRSAVAKNC(5) CTKRNAPDC(3) 4 Phage display (in vivo) 3 PMID:18692101 Ischemic stroke tissue from transient middle cerebral artery occlusion model Not determined. Not determined. Not determined. CX7C T7 phage display library Phage library displaying CX7C random peptides on the viral surface was injected into the tail veins of Adult male Sprague-Dawley rats at 4h following ischemic injury and allowed to circulate for 2h. The ipsilateral (ischemic) hemisphere of each brain was removed and the tissue-bound phages were recovered and amplified for the next round of screening. After three rounds of screening, the titers of recovered phages were enriched by approximately 60-fold. Thirty phage clones from the second and third round were randomly picked and sequenced. 473 NPVENMMDRDSQ(32) NPVEWFMSTVNT(16) NPVESWLTHTRI(10) NPVEHAVNALRP(2) NPVETYTGLYHV(2) NPVEGMLTVLAR(1) NPVEIALTVPTR(1) NPVEHFLSAKLW(1) NELVIPNITPAR(1) NPVE 9 Phage display (subtractive panning) 3 PMID:18715645 FVIII inhibitor positive plasma samples before rituximab treatment Coagulation factor VIII Not determined. Not determined. Ph.D.-12 phage display library (X12) The FVIII inhibitor titer of the target is 23 BUs/ml ( BU = Bethesda Unit, corresponds to 50% reduction of FVIII activity). An IgG preparation mixed 1:1 with plasma of a healthy donor was used in 2 negative selections to remove non-specific phages. For 3 rounds of positive selection, binding phages were eluted by pH-shift using glycine-HCl, pH 2.2. Although 300 phage clones were analyzed, only above sequences were given in the original paper. The template of these peptides might be A2 domain of factor VIII. 474 NPIEALM(1) NPVETQV(1) NPVEMLL(1) NPVEHMM(1) NPVESLL(1) NPVE 5 Phage display (subtractive panning) 3 PMID:18715645 FVIII inhibitor positive plasma samples before rituximab treatment Coagulation factor VIII Not determined. Not determined. Ph.D.-7 phage display library (X7) The FVIII inhibitor titer of the target is 23 BUs/ml ( BU = Bethesda Unit, corresponds to 50% reduction of FVIII activity). An IgG preparation mixed 1:1 with plasma of a healthy donor was used in 2 negative selections to remove non-specific phages. For 3 rounds of positive selection, binding phages were eluted by pH-shift using glycine-HCl, pH 2.2. Although 300 phage clones were analyzed, only above sequences were given in the original paper. The template of these peptides might be A2 domain of factor VIII. 475 CPEVDRATC(1) 1 Phage display (subtractive panning) 3 PMID:18715645 FVIII inhibitor positive plasma samples before rituximab treatment Coagulation factor VIII Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The FVIII inhibitor titer of the target is 23 BUs/ml ( BU = Bethesda Unit, corresponds to 50% reduction of FVIII activity). An IgG preparation mixed 1:1 with plasma of a healthy donor was used in 2 negative selections to remove non-specific phages. For 3 rounds of positive selection, binding phages were eluted by pH-shift using glycine-HCl, pH 2.2. Although 300 phage clones were analyzed, only above sequences were given in the original paper. The template of these peptides might be A2 domain of factor VIII. 476 NMIESMLRTASH(1) NPVEWFMSTVNT(1) NPIESMLRTASH(1) NPVE 3 Phage display (subtractive panning, competitive panning) 3 PMID:18715645 FVIII inhibitor positive plasma samples before rituximab treatment Coagulation factor VIII Not determined. Not determined. Ph.D.-12 phage display library (X12) The FVIII inhibitor titer of the target is 23 BUs/ml ( BU = Bethesda Unit, corresponds to 50% reduction of FVIII activity). An IgG preparation mixed 1:1 with plasma of a healthy donor was used in 2 negative selections to remove non-specific phages. For 3 rounds of positive selection, binding phages were eluted by competition with FVIII in excess. Although 300 phage clones were analyzed, only above sequences were given in the original paper. The template of these peptides might be A2 domain of factor VIII. 477 NPVELLL(1) NPVELGI(1) NPVE 2 Phage display (subtractive panning, competitive panning) 3 PMID:18715645 FVIII inhibitor positive plasma samples before rituximab treatment Coagulation factor VIII Not determined. Not determined. Ph.D.-7 phage display library (X7) The FVIII inhibitor titer of the target is 23 BUs/ml ( BU = Bethesda Unit, corresponds to 50% reduction of FVIII activity). An IgG preparation mixed 1:1 with plasma of a healthy donor was used in 2 negative selections to remove non-specific phages. For 3 rounds of positive selection, binding phages were eluted by competition with FVIII in excess. Although 300 phage clones were analyzed, only above sequences were given in the original paper. The template of these peptides might be A2 domain of factor VIII. 478 NPVEALLRPLGS(1) NPVENMMDRDSQ(1) NPVERLLTSALA(1) NPVEWLMSTVNT(1) NPVE 4 Phage display (subtractive panning) 3 PMID:18715645 FVIII inhibitor positive plasma samples after rituximab treatment Coagulation factor VIII Not determined. Not determined. Ph.D.-12 phage display library (X12) The FVIII inhibitor titer of the target is 23 BUs/ml ( BU = Bethesda Unit, corresponds to 50% reduction of FVIII activity). An IgG preparation mixed 1:1 with plasma of a healthy donor was used in 2 negative selections to remove non-specific phages. For 3 rounds of positive selection, binding phages were eluted by pH-shift using glycine-HCl, pH 2.2. Although 300 phage clones were analyzed, only above sequences were given in the original paper. The template of these peptides might be A2 domain of factor VIII. 479 NPVEHMM(2) NPVENLT(1) NPVETQV(1) NPVE 3 Phage display (subtractive panning) 3 PMID:18715645 FVIII inhibitor positive plasma samples after rituximab treatment Coagulation factor VIII Not determined. Not determined. Ph.D.-7 phage display library (X7) The FVIII inhibitor titer of the target is 23 BUs/ml ( BU = Bethesda Unit, corresponds to 50% reduction of FVIII activity). An IgG preparation mixed 1:1 with plasma of a healthy donor was used in 2 negative selections to remove non-specific phages. For 3 rounds of positive selection, binding phages were eluted by pH-shift using glycine-HCl, pH 2.2. Although 300 phage clones were analyzed, only above sequences were given in the original paper. The template of these peptides might be A2 domain of factor VIII. 480 NPIESMLRTASH(1) NPIEQLLRASYN(1) NPIENALGVREI(1) NPIE 3 Phage display (subtractive panning, competitive panning) 3 PMID:18715645 FVIII inhibitor positive plasma samples after rituximab treatment Coagulation factor VIII Not determined. Not determined. Ph.D.-12 phage display library (X12) The FVIII inhibitor titer of the target is 23 BUs/ml ( BU = Bethesda Unit, corresponds to 50% reduction of FVIII activity). An IgG preparation mixed 1:1 with plasma of a healthy donor was used in 2 negative selections to remove non-specific phages. For 3 rounds of positive selection, binding phages were eluted by competition with FVIII in excess. Although 300 phage clones were analyzed, only above sequences were given in the original paper. The template of these peptides might be A2 domain of factor VIII. 481 NPVEHMM(1) NPVELGI(1) NPVEFHT(1) NPVESLL(1) NPVE 4 Phage display (subtractive panning, competitive panning) 3 PMID:18715645 FVIII inhibitor positive plasma samples after rituximab treatment Coagulation factor VIII Not determined. Not determined. Ph.D.-7 phage display library (X7) The FVIII inhibitor titer of the target is 23 BUs/ml ( BU = Bethesda Unit, corresponds to 50% reduction of FVIII activity). An IgG preparation mixed 1:1 with plasma of a healthy donor was used in 2 negative selections to remove non-specific phages. For 3 rounds of positive selection, binding phages were eluted by competition with FVIII in excess. Although 300 phage clones were analyzed, only above sequences were given in the original paper. The template of these peptides might be A2 domain of factor VIII. 482 MPPPLMQ FHENWPS 2 Phage display (common panning) 4 PMID:17972282 Carbon Black FW-18 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 483 RTAPTTPLLLSL WHLSWSPVPLPT 2 Phage display (common panning) 4 PMID:17972282 Carbon Black FW-18 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 484 VPRVTSI MANHNLS FHENWPS 3 Phage display (common panning) 4 PMID:17972282 Long fibrous cellulose Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage ELISA showed that FHENWPS did not show specific binding activity to cellulose. 485 THKTSTQRLLAA KCCYVNVGSVFS AHMQFRTSLTPH 3 Phage display (common panning) 4 PMID:17972282 Long fibrous cellulose Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage ELISA showed that AHMQFRTSLTPH did not show specific binding activity to cellulose. 486 WYRGRL(94) DPHFHL(1) RVMLVR(1) 3 Phage display (common panning) 5 PMID:18246072 Denuded cartilage grafts Not determined. Not determined. Not determined. f3-6mer phage display library (X6) WYRGRL was selected in 94 of 96 clones sequenced after five rounds of biopanning and was demonstrated to bind to collagen IIα1. Peptide-functionalized nanoparticles targeted articular cartilage up to 72-fold more than nanoparticles displaying a scrambled peptide sequence following intra-articular injection in the mouse. 487 ATETLARSLRLF(1) YKHGMVTVGSTP(1) 2 Phage display (common panning) 2 PMID:18952877 Magnetic particles (BacMPs) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The BacMPs obtained from Magnetospirillum magneticum strain AMB-1 consist of pure magnetite (50 to 100 nm in size) and are covered with a lipid bilayer membrane derived from the invagination of the inner membrane. Forty-six clones were randomly picked from positive phages and sequenced. 488 KSLSRHDHIHHH(2) HQTVVRPIPLFR(1) ASHMSWLGPGLR(1) SSLYPARLQGMS(1) 4 Phage display (common panning) 3 PMID:18952877 Magnetic particles (BacMPs) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The BacMPs obtained from Magnetospirillum magneticum strain AMB-1 consist of pure magnetite (50 to 100 nm in size) and are covered with a lipid bilayer membrane derived from the invagination of the inner membrane. Fourteen clones were randomly picked from positive phages and sequenced. 489 KPQQHNRPLRHK KIPHPEHPTKFR VFAGKPSHKPPH 3 Phage display (common panning) 4 PMID:18952877 Magnetic particles (BacMPs) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The BacMPs obtained from Magnetospirillum magneticum strain AMB-1 consist of pure magnetite (50 to 100 nm in size) and are covered with a lipid bilayer membrane derived from the invagination of the inner membrane. Twenty-four clones were randomly picked from positive phages and sequenced. 490 QFNVQKVPKSKP(2) GPVHKHLPKAHK(1) 2 Phage display (common panning) 5 PMID:18952877 Magnetic particles (BacMPs) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The BacMPs obtained from Magnetospirillum magneticum strain AMB-1 consist of pure magnetite (50 to 100 nm in size) and are covered with a lipid bilayer membrane derived from the invagination of the inner membrane. Twenty-four clones were randomly picked from positive phages and sequenced. The peptides given in the original articles were highly cationic and could bind onto the BacMP membrane. They exhibited antimicrobial activity against Bacillus subtilis but not against Escherichia coli and Saccharomyces cerevisiae. 491 GPVHKHLPKAHK(3) KPIHHHPHLPLK(2) KPQQHNRPLRHK(2) KIPHPEHPTKFR(2) VFAGKPSHKPPH(2) 5 Phage display (common panning) 6 PMID:18952877 Magnetic particles (BacMPs) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The BacMPs obtained from Magnetospirillum magneticum strain AMB-1 consist of pure magnetite (50 to 100 nm in size) and are covered with a lipid bilayer membrane derived from the invagination of the inner membrane. Forty-eight clones were randomly picked from positive phages and sequenced. The peptides given in the original articles were highly cationic and could bind onto the BacMP membrane. They exhibited antimicrobial activity against Bacillus subtilis but not against Escherichia coli and Saccharomyces cerevisiae. 492 ETPLTETALKWH(29) FNGSHIYSPFHP(7) HLQDGSPPSSPH(6) APVPPTAWWHLS(5) QWNLTPRQSLQL(4) 5 Phage display (common panning) 3 PMID:19226949 Anti-H5N1 virus monoclonal antibody 8H5 Hemagglutinin Not determined. Not determined. Ph.D.-12 phage display library (X12) Sixty positive clones were sequenced. ETPLTETALKWH was further displayed on the virus-like particle assembled from aa 1-149 fragment of HBcAg. The chimeric particle conserved the specific binding to 8H5 mAb, and competed with H5N1 viruses for 8H5 mAb. 493 VGPRR FLLCEQ RVPMAR RWPELE PRVKGA ADGRAV GMVGQG HGRKRR GSWSSM RARATM LWRGPK AVVLLS SRGRLG GYGVDA VRSLIF RDRLPP PGSRER QVDQGS GRVNG PERCWM RCITAR 21 Phage display (common panning) 5 PMID:18979633 Implantation serine proteinase complex (Serine protease 28, 29) Not determined. Not determined. Not determined. X6 T7 phage display library 494 SRARSA(2) LRAAFQ(1) VLWTLR(1) KDLLKC(1) GGFTHV(1) YRSVEW(1) RTATGR(1) KRSTVR(1) LRRGGV(1) VAARSA(1) VMVRSV(1) MTFRSA(1) ARSIRV(1) GESTHG(1) VEVAKD(1) WFDNTM(1) QRGVLR(1) ARRWRR(1) RARLRQ(1) SGWRVG(1) RFRQKF(1) WKRQRW(1) NRRSWK(1) ERSRRS(1) GGWRKA(1) YNRMAG(1) YSSKRA(1) RRRGNG(1) LGVRAR(1) QVTRKV(1) ARGSRG(1) 31 Phage display (common panning) 5 PMID:18979633 Kallikrein-6 Not determined. Not determined. Not determined. X6 T7 phage display library 495 LDVVLAWRDGLSGAS GVVWRYTAPVHLGDG 2 Phage display (common panning) 3 PMID:18157673 Monoclonal antibody ME361 GD2 ganglioside Not determined. Not determined. fUSE5-based X15 phage display library The peptides inhibited binding of the mAb to GD2. When coupled to keyhole limpet hemocyanin (KLH) or presented as multiantigenic peptides in QS21 adjuvant, the peptides induced in mice antibodies binding specifically to GD2 and delayed-type hypersensitive lymphocytes reactive specifically with GD2-positive D142.34 mouse melanoma cells. The immunity elicited by the peptides significcantly inhibited growth of GD2-positive melanoma cells in mice. 496 NVVRQ(2) 1 Bacterial display (subtractive panning) 4 PMID:18765541 Highly metastatic prostate carcinoma cell line PC-3M-1E8 Not determined. Not determined. Not determined. FliTrx bacterial display library (X12) The peptides were selected in vitro by screening of the FliTrx library with the highly metastatic prostate carcinoma cell line PC-3M-1E8 for four rounds. For each round, 1e8 FliTrx cells were added to a cell culture dish with the non-metastatic human prostate cancer cell line, PC-3M-2B4, for negative selection, and then incubated for 1 h with PC-3M-1E8 cells for positive selection. After four rounds, 100 individual FliTrx clones were selected and their peptide-encoding inserts were sequenced and analyzed for potential repetitive peptide motifs. However, only one sequence NVVRQ was given in the original paper. NVVRQ-displaying FliTrx clone 27 or 50 specifically bound to PC-3M-1E8 cells 12.6 or 11.7 times over control PC-3M-2B4 cells, respectively. NVVRQ specifically bound to a series of highly metastatic tumor cells, including prostate cancer PC-3M-1E8, breast cancer MDA-MB-435S, lung cancer PG-BE1, and gastric cancer MKN-45sci, in vitro and in vivo but not to the poorly metastatic or non-metastatic cell line, including prostate cancer PC-3M-2B4, breast cancer MCF-7, lung cancer PG-LH7, or murine fibroblast cell NIH/3T3. FITC-NVVRQ strongly and specifically targeted the metastasis foci in tumor-bearing mice 24 h after i.v. peptide injection. 497 CHAQGSAEC(11) 1 Phage display (in vivo) 3 PMID:18978306 Mouse thymus vasculature Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Eleven clones were sequenced. All of them has the same sequence CHAQGSAE.Immunohistochemistry confirmed that the phage peptide CHAQGSAEC can bind specifically to thymus blood vessels in mice. Furthermore, phage peptide CHAQGSAEC and free peptide CHAQGSAEC can inhibit the bioactivity of thymus output in vivo. 498 CEMQTSTAC(1) CHSSPTLYC(1) CQSEVSQLC(1) CPLITAAFC(1) CLGNKAHTC(1) CNKGAGKYC(1) CSPDLPQRC(1) CGTNKPWNC(1) CMPNTLREC(1) CTPRADRHC(1) CTQHNPHQC(1) 11 Phage display (in vivo) 3 PMID:18978306 Mouse kidney Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Eleven clones were sequenced. 499 NSLSNASEFRAP(6) 1 Phage display (common panning) 5 PMID:18330476 Angiopoietin-1 receptor Angiopoietin-1, ANG-1 4K0V , Not determined. Ph.D.-12 phage display library (X12) Phages bound to Tie2 receptors were eluted with angiopoietin-2. After five rounds of screening, 20 phage clones were picked out randomly and amplified for DNA sequencing. Of these, only 17 clones had efficiently inserted peptides, and approximately 35% (6/17) of recovered clones expressed the consensus amino acid sequence NSLSNASEFRAP. Binding assays and Scatchard analysis revealed that NSLSNASEFRAP could specifically bind to Tie2 with a dissociation constant of 2.1×10−8 M. In addition, it wa showed that NSLSNASEFRAP was internalized into tumor cells highly expressing Tie2. Another enriched phage clone displayed a peptide XXGTHGHCQLSH, which did not have binding specificity for Tie2. 500 QMDTSTSLAPSR(1) VPFTLQTRSLSD(1) TVTPSNISFTPS(1) ASTLINPLSISL(1) AGSTASVTPAKH(1) QMANSVMPLSWT(1) YAHSHDKYHPN(1) NQSPHSTYTLKP(1) HNYPQSYRPPIV(1) TDNNTTALTPSH(1) TMNNTTATVSPS(1) FQKQTNQSVSVS(1) VHMTPTNLTPNL(1) TFSYHNSNSPT(1) VPDHQVSYTLSR(1) IFHSHASLSPNS(1) ADNANVSTLHPT(1) VNQQPSSAFSPS(1) LSTVQTISPSNH(1) DMNHTKSSYNPS(1) [ST]-[AVLIFYWFYW]-[ST]-P-[ST] 20 Phage display (common panning) 3 PMID:18546729 Hematite particles Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A search of published sequence data revealed that the binding motif (Ser/Thr-Pro-Ser/Thr) is adjacent to the terminal heme-binding domain of both OmcA and MtrC, which are outer membrane cytochromes from the metal reducing bacterium Shewanella oneidensis MR-1. The entire five amino acid consensus sequence (Ser/Thr-hydrophobic/aromatic-Ser/Thr-Pro-Ser/Thr) was also found as multiple copies in the primary sequences of metal-oxide binding proteins Sil1 and Sil2 from Thalassiosira pseudonana. It is suggested that this motif constitutes a natural metal-oxide binding archetype that could be exploited in enzyme-based biofuel cell design and approaches to synthesize tailored metal-oxide nanostructures. 501 CGRTRVTSC(2) CGRTRDN(2) CRNPKKATC(2) CSTKRKPNC(2) CAREVTLLC(1) CSREVTLLC(1) CDTTIANCC(1) CDRILSPSC(1) CTPKKSGRC(1) CTTSSLTDC(1) CGSNSLTPC(1) CCKSLRPHC(1) CTKPKRNNC(1) 13 Phage display (common panning) 4 PMID:19121687 Anti-LPS monoclonal antibody 9D5 Lipopolysaccharide, LPS Not determined. Not determined. CX7C T7 phage display library 502 CGRTRVTSC(4) CAPRSSNRC(3) CAREVTLLC(2) CGRTRDN(1) CAPQ(1) CLLLAQTDC(1) SKKIPTC(1) CLGRISPPC(1) CASNSLTPC(1) 9 Phage display (common panning) 4 PMID:19121687 Anti-EPS monoclonal antibody 4B11 Exopolysaccharide, EPS Not determined. Not determined. CX7C T7 phage display library 503 NSEQLNFVMKVV(4) GYVGPRLGSGIG(1) TVVMGRVWQYEQ(1) GYVGPRLEVGDW(1) 4 Phage display (common panning) 4 PMID:19121687 Anti-EPS monoclonal antibody 4B11 Exopolysaccharide, EPS Not determined. Not determined. Ph.D.-12 phage display library (X12) 504 CLGRVLANC(3) CPYPRKGSC(2) CVPKKNRTC(2) CGRTRVTSC(2) CGRTRDN(1) CRFLRRTVC(1) CRGRTHPLC(1) CDTKKNHCC(1) CNICARQYC(1) CVRNSLTPC(1) CRGRTLHLC(1) CRGRTHPRC(1) CSYVGKGSC(1) 13 Phage display (common panning) 4 PMID:19121687 Monoclonal antibody BPA Not determined. Not determined. Not determined. CX7C T7 phage display library 505 CGRTRVTSC(2) CGRTRDN(2) CAREVTLLC(2) CGAESLTPC(2) CFTVARACC(2) CGPKRKATC(1) CSGLLVANC(1) CSKKNPGNC(1) CGSESLTPC(1) CCKSLRPHC(1) CAKTRTAKC(1) CRTKKSGTC(1) CKTRKSGSC(1) 13 Phage display (common panning) 4 PMID:19121687 Monoclonal antibody BPM Not determined. Not determined. Not determined. CX7C T7 phage display library 506 RNTRHPPPWSAE WPGSNLTPRTQT LIKTSPRSERYA SVSVGCKPVPRP SHLLKPQTRTRT QLSGTLERQWQR VLPPALEITPLT VIPLAIEFTPRP AQVTLPIRTPNA P-S-R-T-P-R-P-E-W-A-x-L 9 Phage display (common panning) 3 PMID:18242709 Anti-Pru p 3 polyclonal antibody IgE (from OSA-pool) Non-specific lipid-transfer protein 1, LTP 1 Not determined. Not determined. Ph.D.-12 phage display library (X12) The target was purified from pooled sera of 14 peach allergic patients showing only oral allergy syndrome. Twenty clones were Sequenced; 9 different peptides were rendered. 507 LTRGSPVTGPFA SLGAAQVVLPRT NTDSALRLPPFY SLTPLLFNYDVA FHAEWPTPSPWP NDDLMATRHGLA KSNDLGSAPLQH TDHSMGQRVQPS RWWQHRPSHTPS HNTVPSLWYHNR QLSGMLEWQNLR TPTGVHIAPVQT TSRPALLNDQGH 12 Phage display (common panning) 3 PMID:18242709 Anti-Pru p 3 polyclonal antibody IgE (from SYS-pool) Non-specific lipid-transfer protein 1, LTP 1 Not determined. Not determined. Ph.D.-12 phage display library (X12) The target was purified from pooled sera of 9 peach allergic patients with, at least, one systemic reaction, with or without associated oral allergy syndrome. Twenty clones were Sequenced; 12 distinct peptides were rendered. 508 CFRHMTEQC(6) CIPLPFYNC(5) CSHLYLHNC(2) CPLTGTSKC(1) CTLKVRGEC(1) CGFWHTSKC(1) CAWPSKDNC(1) CIPLLFHNC(1) CWGAMVHPC(1) CHASLKHRC(1) CYTPWMPRC(1) CTYLARKGC(1) 12 Phage display (common panning) 4 PMID:19027791 Beta-amyloid protein 42 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Seventy-two clones were isolated arbitrarily from the output of the fourth round of panning. Their individual affinities were evaluated and 22 clones showed 3–7 times higher affinity for Aβ42 as compared to that for BSA were sequenced and 12 peptide inserts were identified. The Kd of candidate phage clones for binding to Aβ42 are in the picomolar range. The specific interaction with amyloid plaques was validated by immunohistochemistry on brain sections. 509 SVSLPYANLATH(11) FRTKPDAQGDTS(1) NHNDFRPMYTWR(1) HYDFSIKETSLQ(1) PTATHRPLATVF(1) 5 Phage display (common panning) 3 PMID:18241959 Anti-catalase monoclonal antibody C001 Catalase Not determined. Not determined. Ph.D.-12 phage display library (X12) Among 34 clones randomly selected, 31 clones were positively recognized by mAb C001 while BSA was used as negative control. Fifteen positive clones were sequenced and 5 unique peptides were indentified. One mimotope (SVSLPYANLATH) showed good match with Catalase protein at 394—405aa and the serum of mice induced by the phage clone clearly recognized Catalase protein. 510 TLSYKLYRHSLL(2) SVEKLVRAGQVS(1) VETKLTRETVWT(1) PVSKLSRASGVG(1) FHARLQRLPPHR(1) FHAYSKLTKSLL(1) SWYKLVHGDAQA(1) SPLKLSRSSMYV(1) TPNKLKKPPVPQ(1) WTQRLVKGAEVG(1) DTERLYRAGQEV(1) K-L-x-R 11 Phage display (common panning) 3 PMID:17978885 Anti-nucleoprotein monoclonal antibody N1H5 Nucleoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) 511 NMFKKEMALSLD(2) RTTDFKREMSTS(1) VPLTFKHEMSLA(1) VLPTFKREMTLV(1) WLSGFKRDMVSS(1) GYAPFKREMFQN(1) PVLVFHREMSLG(1) NHWKMEMSLPSS(1) DYNRFHREYVVS(1) FNPWQWEMTFPT(1) FKREM 10 Phage display (common panning) 3 PMID:17978885 Anti-nucleoprotein monoclonal antibody N4E2 Nucleoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) 512 AKAPLDLPVDGF(2) FRASTDYPVFGF(1) STITDKPVPGFG(1) STISDKPVPGFG(1) HDSFWTHPVPGA(1) D-x-P-V-x-G-F 5 Phage display (common panning) 3 PMID:17978885 Anti-nucleoprotein monoclonal antibody H2F5 Nucleoprotein, Protein N Not determined. Not determined. Ph.D.-12 phage display library (X12) 513 SNFTTQMTFYTG(3) VHNSHASWRYTG(1) VQDNTGLLHYTG(1) TQLMWPNLIYTG(1) ASFELHQWYTAT(1) FTSSNVPLRYTG(1) SVTTLPAQLYTG(1) YTG 7 Phage display (common panning) 3 PMID:17978885 Anti-nucleoprotein monoclonal antibody H3G5 Nucleoprotein, Protein N Not determined. Not determined. Ph.D.-12 phage display library (X12) 514 MNLHDYHRLFWY QHPQINQTLYRM WWRPLTPESPPA 3 Phage display (common panning) 3 PMID:18359289 Enterotoxin type B Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Among 38 phage clones selected, 16 were positive. Phage clone number 24, 9, and 6 with higher SEB-binding affinity were sequenced. 515 CLSYYPSYC(14) CSSYYPSYC(2) CEQWYPSYC(1) CQHPHKPGC(1) CLEIQPPHC(1) CGAGGSLYC(1) CHPNGDKLC(1) CPNSSLQQC(1) CYTQPLSTC(1) CHLPCSHRC(1) CRHGPTAEC(1) CKNMPPWYC(1) CPTPWMARC(1) CATKTPRTC(1) YYPSY 14 Phage display (subtractive panning) 4 PMID:18363834 L-alpha-phosphatidylserine Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) L-α-phosphatidylcholine (PC) coated and BSA blocked ELISA plates were used to remove non-specific phages. 516 EYWYCGMNRTGC(6) QIWYERTLPFTF(1) 2 Phage display (common panning) 3 PMID:18481004 Anti-blood group A antigen monclonal antibody NaM87-1F6 Blood group A antigen Not determined. Not determined. Ph.D.-12 phage display library (X12) 517 WPMNDDSTRPR(1) DSMNTDTSTRPR(1) FQTDMSTRPRGA(1) YTYTDYSTRPHY(1) DPTDYSTRPHMW(1) PQTDGSTRPRGL(1) DFRYSTRPFAL(1) QRYYSTRPRAL(1) MISYSTRPDRQ(1) VQTDGSTKSHRH(1) IRTDESTKRLPP(1) TLSTDQSTTRNW(1) DIDYSDYSTR(1) DDHSTRCWVS(1) YPLHLNTDYSTR(1) T-D-x-S-T-R-P 15 Phage display (common panning) 4 PMID:19741295 Anti-HSP90 monoclonal antibody AC88 Heat shock protein HSP90 Not determined. Not determined. Ph.D.-12 phage display library (X12) 518 WPMNDDSTRR(5) YTYTDYSTRPHY(3) FSTDWSTRPRLV(1) DSMNTDDSTRPR(1) DFRDYSTRPFAL(1) MISDYSTRPDRR(1) WTRDNSTRPIVR(1) TDSDYSTRPSRG(1) TYQDFSTRPRSF(1) AESDDSSTRRH(1) QFTGSTRQHAG(1) WNTDFSTRAAHT(1) AYKLPQTDDST(1) T-D-x-S-T-R-P 13 Phage display (common panning) 5 PMID:19741295 Anti-HSP90 monoclonal antibody AC88 Heat shock protein HSP90 Not determined. Not determined. Ph.D.-12 phage display library (X12) 519 FPSHYWLYPPPT(7) NYKSPLFAVPMT(6) DLNTNRTQMVLH(4) KIVMFWHEPVYA(4) 4 Phage display (common panning) 4 PMID:19741295 Anti-DnaK monoclonal antibody 8E2/2 Heat shock protein DnaK Not determined. Not determined. Ph.D.-12 phage display library (X12) 520 FPSHYWLYPPPT(11) NYKSPLFAVPMT(5) DLNTNRTQMVLH(3) 3 Phage display (common panning) 5 PMID:19741295 Anti-DnaK monoclonal antibody 8E2/2 Heat shock protein DnaK Not determined. Not determined. Ph.D.-12 phage display library (X12) 521 TISNRDYIRPMD(3) FPSHYWLYPPPT(2) SQMPEYLLKADN(1) WPMNTDDSTRPR(1) YTYTDYSTRPHY(1) PQTDGSTRPRGL(1) TLSQDQSETLNY(1) PHPSTMFDRQED(1) TSPHKTTLDLNA(1) DNHSPVNIAHKL(1) SRLPLSQPSPNS(1) 11 Phage display (common panning) 4 PMID:19741295 Pooled typhoid patients' sera (TPS) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 522 TISNRDYIRPMD(6) SQMPEYLLKADN(3) WPMNTDDSTRPR(1) YTYTDYSTRPHY(1) PQTDGSTRPRGL(1) DPTDYSTRPHMW(1) 6 Phage display (common panning) 5 PMID:19741295 Pooled typhoid patients' sera (TPS) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 523 VTWTPQAWFQWV(4) MTTLSDSPVRPR(2) LTSQEMAARTLG(1) VAPFWVASLPAP(1) YTKPMGLTFPS(1) FSAHAHL(1) 3 Phage display (subtractive panning) 4 PMID:18632177 U87MG glioblastoma cell line Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage clones expressing VTWTPQAWFQWV bound to U87MG cells 700-fold more efficiently than the original unselected library. They bound strongly to other human glioma cell lines, including H4, SW1088 and SW1783, but very weakly to normal human astrocytes and SV40-immortalized human astroglial cells. When compared to other non-glial tumor cells, the phage showed 400- to 1400-fold higher binding efficiency for U87MG cells. 524 YTRISNDHHCDTTICHINST(3) NDSNCTDTLCYVYPLHSNPG(2) VKDALHHVDHTCSTFLCALP(2) DTHFCDDNYCPLPHSSGAIA(2) SCNDYVCLTPHPKTPLYEKQ(1) NVNCNASVCTIPDRLITDNP(1) SSYCNNSLCLVPSYPDHSNT(1) HCNAHLCNLPDGSPSSISSH(1) NIHHCNHIYCTIPSHS(1) IHSYSYASDNMCTDTVCFIH(1) CSDTYCTIPTYGRDRDVRRD(1) DSYFCDNFLCRLDHNRNSRN(1) HNISCDKSLCMLRESAPTTV(1) NSSGSCDHFLCIIPTSNTHT(1) HYSSASHANNVCDAHYCLLP(1) AFCDISFCHLNDRIIIIITN(1) GLDPYHRFCDDNVCSIRPFN(1) DSSDDRHTCGHHICHLYPFS(1) HPPSHYCDIHSRLCHPNAPD(1) C-[DNSTG]-D-[ST]-x-C-x(2)-P 19 Phage display (common panning) 3 PMID:18672101 Thioredoxin-1, Trx-1 Not determined. Not determined. Not determined. ANL 22 phage display library Biotinylated thioredoxin-1 imobilized with streptavidin-coated magnetic beads were used as target. After three rounds of affinity selection, 96 phage clones were screened using a phage enzyme-linked immunosorbent assay (ELISA). A total of 24 phage clones bound specifically to E. coli Trx. 525 YVYRWVEAECVA YVYEGRSRVRRP WHVPRTWWVLPP NTLSGQYFWRMP REPIYHKLHRLT FSPAYLRDAALK YIDHIVLTIYLE 7 Phage display (common panning) 3 PMID:18812010 Anti-cathepsin L1/L2 IgG Cathepsin L1, Cathepsin L2 Not determined. Not determined. Ph.D.-12 phage display library (X12) YVYRWVEAECVA and FSPAYLRDAALK bearing phages had been successfully used in vaccination. 526 KHLPLYR(10) KHNWPPP(8) FHEPKYR(7) GHIAKYR(6) GHLWKYV(2) HLP 5 Phage display (subtractive panning) 3 PMID:18725193 Omalizumab Ig epsilon chain C region Not determined. Not determined. Ph.D.-7 phage display library (X7) In the second and third round of selection, the library was precleared on trastuzumab. 527 PLLQATL(2) RKPGKPV(1) TLHSAQA(1) HNRPRNN(1) RHTHRSH(1) TAPGVST(1) NLTLAWR(1) HTTHMYL(1) WSAPVPN(1) SFRPTPP(1) 10 Phage display (common panning) 3 PMID:20414975 Heparin-binding growth factor 2, HBGF-2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) PLLQATL showed high homology to the immunoglobulin-like (Ig-like) domain III (D3) of high-affinity bFGF receptors, FGFR1 (IIIc) and FGFR2 (IIIc). Synthetic PLLQATL peptides mediate strong inhibition of bFGF-induced cell proliferation and neovascularization. 528 CAWYEWPC(12) CAWYQFPC(6) CVWWQWPC(1) CPWFQWPC(1) CKWFQWPC(7) CFWVNTDC(1) CLYLSIRC(1) x-W-[FYW]-[QE]-[WF]-P 7 Phage display (common panning) 3 PMID:18957413 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. Cys6 phage display library For the first round, FcγRIIA-R134-GST immoblized by precoated with goat anti-GST polyclonal antibody was used for panning. For other rounds, MaxiSorp strips coated FcγRIIA-R134-GST was used. Among 29 selected clones, seven different amino acid sequences were found. 529 CCSVRGSAWAC(2) CILTIHGPLQC(2) CGARLAMAVAC(1) CRDCVVACLGC(1) CSMGLGGTSLC(1) CGAPNLSRLLGC(1) CGLGYRTAHIC(1) CTLRLGVGVRC(1) CHPHFPWATSC(1) 9 Phage display (common panning) 3 PMID:18957413 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. Cys9 phage display library For the first round, FcγRIIA-R134-GST immoblized by precoated with goat anti-GST polyclonal antibody was used for panning. For other rounds, MaxiSorp strips coated FcγRIIA-R134-GST was used. 530 WAWVWLTETAV(23) AVTFKFTGTDL(2) GSSHASLRYPA(1) LLSFAGRSPSC(1) LSGRSSGWRFS(1) RLRFVVHESSG(1) CPLGLLIHTSC(1) 7 Phage display (common panning) 3 PMID:18957413 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. NNK11 phage display library For the first round, FcγRIIA-R134-GST immoblized by precoated with goat anti-GST polyclonal antibody was used for panning. For other rounds, MaxiSorp strips coated FcγRIIA-R134-GST was used. 531 VGGCPWFQWPCKGQ(2) DQECPWFQWPCGAA(2) TRVCPWFQWPCVTG(2) VMKCPWFQWPCDAL(1) RVRCPWFQWPCGMH(1) SRSCPWFQWPCGSV(1) TPNCPWFQWPCLKS(1) 7 Phage display (common panning) 3 PMID:18957413 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. Evo1 phage display library For the first round, FcγRIIA-R134-GST immoblized by precoated with goat anti-GST polyclonal antibody was used for panning. For other rounds, MaxiSorp strips coated FcγRIIA-R134-GST was used. 532 TDRMCRWFQWPC(1) NSRDCAWFQWPC(1) GEDRCLWFQWPC(1) NKDECRWFQWPC(1) IDSRCHWFQWPC(1) GGMKCWWFQWPC(1) GCNACAWFQWPC(1) 7 Phage display (common panning) 3 PMID:18957413 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. Evo2 phage display library For the first round, FcγRIIA-R134-GST immoblized by precoated with goat anti-GST polyclonal antibody was used for panning. For other rounds, MaxiSorp strips coated FcγRIIA-R134-GST was used. 533 CPWFQWSDSGCS(4) CPWFQWPCLSHA(2) CPWFQWPCGARV(2) CPWFQWMLGCV(1) 4 Phage display (common panning) 3 PMID:18957413 Low affinity immunoglobulin gamma Fc region receptor II-a, IgG Fc receptor II-a Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) 3RY6, Not determined. Evo3 phage display library For the first round, FcγRIIA-R134-GST immoblized by precoated with goat anti-GST polyclonal antibody was used for panning. For other rounds, MaxiSorp strips coated FcγRIIA-R134-GST was used. 534 TDTCLMLPLLLGCDEE DPICWYFPRLLGCTTL WYPCYIYPRLLGCDGD GNICMLIPGLLGCSYE VNSCLLLPNLLGCGDD TPVCILLPSLLGCDTQ TVLCSLWPELLGCPPE TFSCLMWPWLLGCESL FGTCYTWPWLLGCEGF SLFCRLLLTPVGCVSQ HLLVLPRGLLGCTTLA TSLCSMFPDLLGCFNL SHPCGRLPMLLGCAES TSTCSMVPGPLGAVSTW KDPCTRWAMLLGCDGE IMTCSVYPFLLGCVDK IHSCAHVMRLLGCWSR T-x(2)-C-x(2)-P-x-L(2)-G-C-x(2)-E 17 Phage display (common panning) 3 PMID:18957574 High affinity immunoglobulin gamma Fc receptor I, IgG Fc receptor I Ig gamma-1 chain C region Not determined. Not determined. CPEP-8 phage display library (X3CX8CX3) These peptides were able to inhibit IgG1 binding to FcγRI. In soluble form, these peptides antagonize superoxide generation mediated by IgG1. In complexed form, they trigger phagocytosis and a superoxide burst. Unlike IgG, these peptides are strictly FcγRI-specific among the FcγRs. 535 HSEAETGPP[2.16 ± 0.18] 1 Phage display (common panning) 3 PMID:19480736 Serum of a pars planitis patient Proline-rich transmembrane protein 2 Not determined. Not determined. J404 phage display library (X9) ELISA Absorbance was measured at 492 nm. Data were reproduced from the graph. One hundred clones were randomly selected from a phage display library after three panning rounds using serum proteins from a PP patient. The immunologic response level of 100 clones selected were determined with a major number of patients, only the clone 83 was recognized stronger in PP patients sera than in healthy sera (PP vs. healthy; P<0.05). An identical amino acid sequence to HSEAETGPP is found in the human proline-rich transmembrane protein 2 which has not been related with eye diseases. 536 SSVLYGGPPSAA[0.65] LPQNVWLHGWHT[0.46] 2 Phage display (common panning) PMID:19299727 Anti-hstone H1 monoclonal antibody 16G9 Histones H1 from calf thymus Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Specific binding of 16G9 mAb to KLH-conjugated SSVLYGGPPSAA and LPQNVWLHGWHT were determined by ELISA. KLH was used as an internal control. Increasing amounts (0–0.5 μg/ml) of 16G9 mAb were added to the wells. Bound 16G9 mAb was detected using biotin-conjugated anti-mouse IgM Ab. Peroxidase-conjugated streptavidin was added and the streptavidin-biotinylated peroxidase complex was detected by ABTS substrate solution. Multiskan Ascent was used to determine absorbance at 405 nm. The A405 values of the binding of 16G9 in the concentration of 0.5 μg/ml to the peptides were shown and the data were reproduced from the graph. The binding of 16G9 mAb to histone H1 was inhibited by SSVLYGGPPSAA, which was also recognized by rat tolerogenic post-orthotopic liver transplantation serum. The binding of 16G9 mAb to SSVLYGGPPSAA was inhibited by histone H1. Abs induced by SSVLYGGPPSAA immunization inhibited Con A-stimulated splenocyte proliferation, and the inhibition was neutralized by preincubation with SSVLYGGPPSAA. Splenocytes stimulated by anti-CD3 Ab were inhibited by SSVLYGGPPSAA-induced Abs. SSVLYGGPPSAA immunization in rats before heterotopic heart transplantation resulted in significant prolonged allograft survival. 537 CPSGHTKAC(2) CTPGKPHSC(2) CGTHSSRIC(1) CLGTQNKEC(1) CKAASANIC(1) CLPTRHMAC(1) CLSAVHNMC(1) 7 Phage display (subtractive panning) 4 PMID:18985310 CD40 ligand, CD40-L Tumor necrosis factor receptor superfamily member 5 3QD6, Not determined. Ph.D.-C7C phage display library (CX7C) The immobilized recombinant hCD154-muCD8 was used as the target. To eliminate potential non-specific binding of phage clones with affinity for plastic or muCD8, two negative selections were performed with plastic and a non-related recombinant fusion protein hCD153-muCD8. Nine phage clones were selected for the ability to bind CD154 expressed on the surface of J558L cells transfected with human CD154. From the nine selected phage clones, seven different amino acidic sequences were obtained and synthesized. All the peptides specifically bound CD154 expressed on J558L. However, only the peptide CLPTRHMAC was found to recognize the active binding site of CD154, as it competed with the blocking anti-CD154 antibody. 538 VQATQSNQHTPR(2) VQATPRLQHTPR(1) VQATTVQIQHAP(1) VQAIQSNQLTPR(1) VQAGQSNAHTAG(1) VQARQSNQHTPR(1) VQNYQSNQHTPR(1) AQALGLSAISPR(1) AQGPPSKQHSPP(1) EQATPRNHNSPP(1) TFATQSNQHTPR(1) VQATQSNQHTPR 11 Phage display (common panning) 4 PMID:19284521 Agglutinin N-acetylgalactosamine Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage particles that were eluted from the lectin with free GalNAc were considered to have been bound to a GalNAc-binding site. 539 SVSVGMKPSPRP(0.75) NHNTSTWAAYST(0.125) TLPSPHSLLTVH(0.125) 3 Phage display (common panning) 3 PMID:19634182 Semiconductor crystalline surface GaAs(100) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 540 SSMEPDPFLALY(0.666) SSVEPGPFLALY(0.111) SSMYPELFLALY(0.111) SSMNPELFLALY(0.111) 4 Phage display (common panning) 3 PMID:19634182 Semiconductor crystalline surface InAs(100) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 541 SVSVGMKPSPRP(0.5) MSGDIISLAPTG(0.125) GPFFPKSLTTTS(0.125) NAPLSHIPENPR(0.125) SISAMPAPANSS(0.125) 5 Phage display (common panning) 3 PMID:19634182 Semiconductor crystalline surface GaN(0001) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The affinity of the SVSVGMKPSPRP peptide towards the GaN(0001) surface is drastically diminished in the presence of water, whereas when PBST has been used as solvent, the adhesion of this peptide was demonstrated. Influence of the eluent proticity and dielectric constant were confirmed by the use of various solvents. The MALDI signal of the peptide adsorbed onto the GaN(0001) decreases in intensity when the dielectric constants of the employed solvents increase. The media with a high dielectric constant reduces the force acting between the peptide and the GaN(0001) and the molecules are washed out of the surface. When rinsing with 1M NaCl directly or after a previous water washing, there was no peptide left on the surface, indicating towards the electrostatic nature of the interactions between the peptide and the GaN(0001) surface. 542 LLADTTHHRPWT(0.444) SVSVGMKPSPRP(0.111) HHLSXKTVGAST(0.111) HDSLLHPARSAP(0.111) FAPEDLPNYPQR(0.111) HSKPQQPPFVXS(0.111) 6 Phage display (common panning) 3 PMID:19634182 Semiconductor crystalline surface ZnSe(100) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 543 SVSVGMKPSPRP(0.285) LVKDIRGAIPYM(0.142) NNQLSFPAGTTR(0.142) HPQNTFAANLRP(0.142) TSANGKPPALML(0.142) SNIAPGVLPRST(0.142) 6 Phage display (common panning) 3 PMID:19634182 Semiconductor crystalline surface ZnTe(100) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 544 APWHFTRVPALV(0.2) INPMVMSNPNNR(0.2) STFSTNWPSTPT(0.2) TTTNWQLSAPAP(0.2) APQHMIWPKPTA(0.2) 5 Phage display (common panning) 3 PMID:19634182 Semiconductor crystalline surface GaAs(111)A Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 545 SVSVGMKPSPRP(0.666) AQDLNYVRLGPS(0.333) 2 Phage display (common panning) 4 PMID:19634182 Semiconductor crystalline surface GaAs(111)A Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 546 SVSVGMKPSPRP(1.0) 1 Phage display (common panning) 6 PMID:19634182 Semiconductor crystalline surface GaAs(111)A Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 547 DHPTNQLAVPSS(0.125) QHMDKLTHRKPH(0.125) HNIHAITPLTPI(0.125) ASASHLNKRFMT(0.125) STTTSNLLNYRI(0.125) SPLTSPGPHVSS(0.125) HSNPHEAIRASR(0.125) HKHTTTPLFTSR(0.125) 8 Phage display (common panning) 3 PMID:19634182 Semiconductor crystalline surface GaSb(100) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 548 SVSVGMKPSPRP(1.0) 1 Phage display (common panning) 4 PMID:19634182 Semiconductor crystalline surface GaSb(100) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 549 HNQTAPHLPRQS(0.2) SVSVGMKPSPRP(0.8) 2 Phage display (common panning) 6 PMID:19634182 Semiconductor crystalline surface GaSb(100) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 550 AYPQFPNLRTSL(0.142) GTLNSSVEPPLG(0.142) NSNSYRVPHGTM(0.142) IPIGPHIGLVGP(0.142) APESMMIIDPGF(0.142) ISTPLSKSPTRL(0.142) YSPPKTTLPAHS(0.142) 7 Phage display (common panning) 3 PMID:19634182 Semiconductor crystalline surface CdSe(100) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 551 SVSVGMKPSPRP(0.4) VGISKPWAGPSV(0.2) SYTNQIYRQNHP(0.2) YVYVGMKPSPRP(0.2) 4 Phage display (common panning) 4 PMID:19634182 Semiconductor crystalline surface CdSe(100) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 552 SVSVGMKPSPRP(1.0) 1 Phage display (common panning) 6 PMID:19634182 Semiconductor crystalline surface CdSe(100) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 553 CDQRTTRLC(1) CPHDPNHPC(1) CQSQTRNHC(1) CLQDMRQFC(1) CLPTDPIQC(1) CPDHPFLRC(1) CSTRAENQC(1) CPSHLDAFC(1) CKTGHMRIC(1) CVRTPTHHC(1) CSGVINTTC(1) CPLASTRTC(1) CSQFPPRLC(1) CLLNKQNAC(1) CKFPLNAAC(1) CSLTPHRSC(1) CKPWPMYSC(1) CLQHDALNC(1) CNANKPKMC(1) CPKHVLKVC(1) CTPDKKSFC(1) CHGKAALAC(1) CNLMGNPHC(1) CLKNWFQPC(1) CKEYGRQMC(1) CQPSDPHLC(1) CSHLPPNRC(1) 27 Phage display (competitive panning) PMID:19515773 Glycoprotein Gn, glycoprotein G1 Anti-glycoprotein G1 monoclonal antibody 6B9/F5 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In fact, density gradient-purified, UV-treated Andes Virus (ANDV) strain CHI-7913 was used in panning. However, glycoprotein G1 is regarded as the target since the peptides were eluted with a monoclonal antibody against the glycoprotein G1. The glycoprotein G1 locates at 4-545 of the glycoprotein precursor (GI:30313865). The in vitro ANDV infection inhibition rate of phages bearing these peptides were lower than 30%. 554 CSPLLRTVC(1) CHKGHTWNC(1) CINASHAHC(1) CWPPSSRTC(1) CPSSPFNHC(1) CEHLSHAAC(1) CQDRKTSQC(1) CTDVYRPTC(1) CGEKSAQLC(1) CSAAERLNC(1) CFRTLEHLC(1) CEKLHTASC(1) CSLHSHKGC(1) CNSHSPVHC(1) CMQSAAAHC(1) CPAASHPRC(1) CKSLGSSQC(1) 17 Phage display (competitive panning) PMID:19515773 Glycoprotein Gn, glycoprotein G1 Anti-glycoprotein G1 monoclonal antibody 6B9/F5 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In fact, density gradient-purified, UV-treated Andes Virus (ANDV) strain CHI-7913 was used in panning. However, glycoprotein G1 is regarded as the target since the peptides were eluted with a monoclonal antibody against the glycoprotein G1. The glycoprotein G1 locates at 4-545 of the glycoprotein precursor (GI:30313865). The in vitro ANDV infection inhibition rate of phages bearing these peptides were between 30% and 60%. 555 CPSNVNNIC(1) 1 Phage display (competitive panning) PMID:19515773 Glycoprotein Gn, glycoprotein G1 Anti-glycoprotein G1 monoclonal antibody 6B9/F5 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In fact, density gradient-purified, UV-treated Andes Virus (ANDV) strain CHI-7913 was used in panning. However, glycoprotein G1 is regarded as the target since the peptides were eluted with a monoclonal antibody against the glycoprotein G1. The glycoprotein G1 locates at 4-545 of the glycoprotein precursor (GI:30313865). The phage bearing peptide CPSNVNNIC is the most potent phage identified by elution with the anti-Gn antibody 6B9/F5. It inhibited hantavirus entry by greater than 60%. 556 CTTMTRMTC(2) CHPGSSSRC(1) CSLSPLGRC(1) CTARYTQHC(1) CHGVYALHC(1) CLQHNEREC(1) CHPSTHRYC(1) CPGNWWSTC(1) CGMLNWNRC(1) CPHTQFWQC(1) CTPTMHNHC(1) CDQVAGYSC(1) CIPMMTEFC(1) CERPYSRLC(1) CPSLHTREC(1) CSPLQIPYC(1) 16 Phage display (competitive panning) PMID:19515773 Glycoprotein Gc, glycoprotein G2 Anti-glycoprotein G2 monoclonal antibody 6C5/D12 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In fact, density gradient-purified, UV-treated Andes Virus (ANDV) strain CHI-7913 was used in panning. However, glycoprotein G2 is regarded as the target since the peptides were eluted with a monoclonal antibody against the glycoprotein G2. The glycoprotein G2 locates at 653-1137 of the glycoprotein precursor (GI:30313865). The in vitro ANDV infection inhibition rate of phages bearing these peptides were lower than 30%. 557 CNKPFSLPC(1) CHNLESGTC(1) CNSVPPYQC(1) CSDSWLPRC(1) CSAPFTKSC(1) CEGLPNIDC(1) CTSTHTKTC(1) CLSIHSSVC(1) CPWSTQYAC(1) CTGSNLPIC(1) CSLAPANTC(1) CGLKTNPAC(1) CRDTTPWWC(1) CHTNASPHC(1) CTSMAYHHC(1) CSLSSPRIC(1) CVSLEHQNC(1) CRVTQTHTC(1) CPTTKSNVC(1) CSPGPHRVC(1) CKSTSNVYC(1) CTVGPTRSC(1) 22 Phage display (competitive panning) PMID:19515773 Glycoprotein Gc, glycoprotein G2 Anti-glycoprotein G2 monoclonal antibody 6C5/D12 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In fact, density gradient-purified, UV-treated Andes Virus (ANDV) strain CHI-7913 was used in panning. However, glycoprotein G2 is regarded as the target since the peptides were eluted with a monoclonal antibody against the glycoprotein G2. The glycoprotein G2 locates at 653-1137 of the glycoprotein precursor (GI:30313865). The in vitro ANDV infection inhibition rate of phages bearing these peptides were between 30% and 60%. 558 CPMSQNPTC(1) CPKLHPGGC(1) 2 Phage display (competitive panning) PMID:19515773 Glycoprotein Gc, glycoprotein G2 Anti-glycoprotein G2 monoclonal antibody 6C5/D12 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In fact, density gradient-purified, UV-treated Andes Virus (ANDV) strain CHI-7913 was used in panning. However, glycoprotein G2 is regarded as the target since the peptides were eluted with a monoclonal antibody against the glycoprotein G2. The glycoprotein G2 locates at 653-1137 of the glycoprotein precursor (GI:30313865). From phages eluted with the anti-Gc antibody 6C5/D12, those bearing peptide CPMSQNPTC and CPKLHPGGC inhibited hantavirus entry by 66% and 72%, respectively. 559 HSIRNDVRLPSM(7) HSIRTQWTQTQV(4) HSIRQYFTLPAP(3) TNRHNPHHLHHV(3) TPHLQSGFLLTL(3) HISR 5 Phage display (common panning) 3 PMID:19493004 S-ribosylhomocysteine lyase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) TNRHNPHHLHHV was shown to partially inhibit the activity of the LuxS enzyme. Furthermore, 14 peptides containing the consensus sequence HSIR showed high affinity with LuxS. 560 SAMWDF(3)[2.4 ± 0.13, 1.82 ± 0.09] SFIWDF(2)[2.1 ± 0.15, 1.70 ± 0.10] TNMWDF(2)[2.05 ± 0.12, 1.42 ± 0.08] ITMWDF(2)[2.2 ± 0.13, 1.36 ± 0.06] SDWWDF(1)[2.1 ± 0.10, 1.48 ± 0.12] [ST]-x(2)-W-D-F 5 Phage display (common panning) 3 PMID:19118103 Anti-hCXCR1 monoclonal antibody 5A12 C-X-C chemokine receptor type 1, CXC-R1, CXCR-1 Not determined. Not determined. fdMED1-based X6 phage display library ELISA ELISA was used to test the reactivity of mAb 5A12 with phage-peptide clones and the corresponding synthesized peptides, respectively. For phage ELISA, the absorbance was measured at 450 nm (A450 nm). Data shown are A450nm after substraction of background. For peptide ELISA, the absorbance was measured at 450 nm (A450 nm). As control, the linear hexapeptide MRFIAW was used. Data shown were reproduced from the graph. The A450 of MRFIAW is 0.06. The phage bearing the peptides showed specific binding to immobilized mAb 5A12. In FACScan analysis, all selected phage peptides were able to strongly inhibit the binding of mAb 5A12 to hCXCR1-transfected preB 300-19 murine cells. Furthermore, synthetic peptides of the corresponding phage epitopes were effective in blocking the antibody-CXCR1 interactions and to inhibit the binding of hCXCL8 to hCXCR1 transfectants. In vivo, SAMWDF blockade hCXCL8-induced neutrophil recruitment in skin inflammation in rabbits. 561 FWDDFW(4)[2.3 ± 0.15, 1.90 ± 0.15] LWDDFW(2)[1.73 ± 0.12, 1.60 ±0.16] MWNDFW(2)[1.91 ± 0.10, 1.79 ± 0.15] FWLDFW(2)[1.89 ± 0.09, 1.61 ± 0.11] [FLM]-W-x-D-F-W 4 Phage display (common panning) 3 PMID:19118103 Anti-hCXCR2 monoclonal antibody 6C6 C-X-C chemokine receptor type 2, CXC-R2, CXCR-2 Not determined. Not determined. fdMED1-based X6 phage display library ELISA ELISA was used to test the reactivity of mAb 6C6 with phage-peptide clones and the corresponding synthesized peptides, respectively. For phage ELISA, the absorbance was measured at 450 nm (A450 nm). Data shown are A450nm after substraction of background. For peptide ELISA, the absorbance was measured at 450 nm (A450 nm). As control, the linear hexapeptide MRFIAW was used. Data shown were reproduced from the graph. The A450 of MRFIAW is 0.05. The phage bearing the peptides showed specific binding to immobilized mAb 6C6. In FACScan analysis, all selected phage peptides were able to strongly inhibit the binding of mAb 6C6 to hCXCR2-transfected preB 300-19 murine cells. Furthermore, synthetic peptides of the corresponding phage epitopes were effective in blocking the antibody-CXCR2 interactions and to inhibit the binding of hCXCL8 to hCXCR2 transfectants. In vivo, FWDDFW blockade hCXCL8-induced neutrophil recruitment in skin inflammation in rabbits. 562 YTTPPYYVWEWM(1)[1.68] YSPSWDYLTFLM(1)[1.84] YTGQGWQLILPM(1)[0.38] YSEPVSFGWLWM(1)[1.75] YSDMPSDWLFPM(1)[1.84] YGEDANSWFVFM(1)[1.98] YENELGEWWLFM(1)[1.92] YTAPPWNWEWAM(1)[1.94] YQPSSALSSWMM(1)[1.74] YSDTDWMYFSTM(1)[2.05] 10 Phage display (common panning) 5 PMID:19228693 Platelet glycoprotein VI, GPVI Not determined. Not determined. Not determined. Y-X10-M phage display library ELISA Optical density at 450 nm was measured. Data were reproduced from the graph. The human GPVI protein (residues Gln21-Phe234) was expressed as a chimera with a human IgG Fc2 portion at the C terminus using the pcDNA3.1 vector expressed by CHO cells was used as the target. Platelet glycoprotein VI (GPVI) is a major collagen receptor on the platelet surface that recognizes the glycine-proline-hydroxyproline (GPO) sequence in the collagen molecule and plays a crucial role in thrombus formation. These 10 clones were examined for their GPVI-binding ability in phage enzyme-linked immunosorbent assay. A recombinant protein irrelevant to GPVI, which has human IgG-Fc, was used as a control protein. The phage bearing YTGQGWQLILPM bind neithor GPVI-Fc2 nor the control protein. The other nineclones bound to GPVI-Fc2 and did not bind to the control protein. 563 ATWVSPY(6) AHSMGTG(1) FSSQMRY(1) GVGLPHT(1) QIEPLAL(1) RIVLPTY(1) VQQVALL(1) IVLPVPY(1) GHWTRLA(1) NLPLHST(1) 10 Phage display (common panning) 3 PMID:20091097 Commercially pure titanium, cp-Ti Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ATWVSPY exhibited the strongest binding affinity to cp-Ti disks. 564 ATWVSPY(8) GVGLPHT(2) 2 Phage display (common panning) 4 PMID:20091097 Commercially pure titanium, cp-Ti Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ATWVSPY exhibited the strongest binding affinity to cp-Ti disks. 565 QTPLTMAALELF(23) DTPLTTAALRLV(17) ETQLTTAGLRLL(9) ETPLTETALKWH(8) QTPLTETALKWH(4) QTPLTMAALELL(2) HLQDGSPPSSPH(2) GHVTTLSLLSLR(2) ETPLTEPAFKRH(2) T-x-L-T-x(3)-L 9 Phage display (common panning) 3 PMID:18973474 Anti-H5N1 virus monoclonal antibody 8H5 Hemagglutinin Not determined. Not determined. Ph.D.-12 phage display library (X12) A total of 190 randomly picked clones of screened phage were individually tested by ELISA for 8H5 binding. Among them, 69 reactive clones were sequenced. 566 CNDFRSKTC(0.47)[0.87 ± 0.02] CQHSTKWFC(0.105)[2.29 ± 0.05] CLPYAAKHC(0.05)[1.09 ± 0.04] CILGDKVGC(0.05)[1.16 ± 0.06] 4 Phage display (common panning) 4 PMID:19497129 Avian influenza virus (H9N2) particles Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Optical density at 410 nm was measured. Data were reproduced from the graph. After the 4th round, 35 individual clones were sequenced. However, only the sequences above were given in the original paper. Among them, the phage displaying the peptide NDFRSKT possessed good anti-viral properties in vitro and in ovo. This peptide inhibited the hemagglutination activity of the viruses but showed very little and no effect on neuraminidase and hemolytic activities respectively. The phage-antibody competition assay proved that this peptide competed with anti-influenza H9N2 antibodies for the binding sites. Based on yeast two-hybrid assay, we observed that NDFRSKT inhibited the viral replication by interacting with the HA protein and this observation was further confirmed by coimmunoprecipitation. 567 CHPQFLSLC(0.55) CGLYNHPQC(0.27) 2 Phage display (common panning) 3 PMID:19497129 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. Ph.D.-C7C phage display library (CX7C) Twenty clones were sequenced from the 3rd round of panning against Streptavidin. However, only the sequences above were given in the original paper. 568 HYKWLNDPLAAW(1) 1 Phage display (common panning) 4 PMID:19349218 Anti-cEG95 polyclonal antibody EG95 host-protective vaccine antigen Not determined. Not determined. Ph.D.-12 phage display library (X12) Twelve phage clones from the 20 mer library, and eight from the 12 mer library were selected and shown to present different peptides. However, only the peptide E100, i.e. HYKWLNDPLAAWone was given in the original paper. 569 FLLEPHLMDTSM(47) FLLEPHTVTWGA(16) FLIEPWHSSLQS(11) AFLFSPLLAWPT(6) TYRFGPLEPVAF(2) F-L-[LI]-E-P 5 Phage display (subtractive panning) 4 PMID:18563328 Hepatocellular carcinoma cell line HepG2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the process of the second round of panning, the amplified phages were incubated with human liver cell line L02 cells at first to remove phages binding to human liver cells. Among 100 phages that specifically bound to and internalized in HepG2 cells, 82 clones demonstrated highly specific affinity to HepG2 cells and their binding to L02 cells was relatively weak. 570 FKFWLYEHVIRG(6) YWFHNFPTKMYA(4) FYRFVGDHKQLY(4) YIWPLMGSHYAK(2) WHYGVELWIRRF(1) 5 Phage display (common panning) 3 PMID:19056299 96-well immunoassay plate (polystyrene, PS) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) FKFWLYEHVIRG-fused proteins showed a considerably improved affinity to polystyrene microplate, indicating its application in diagnostic technology such as enzyme-linked immunosorbent assay (ELISA). 571 NSLSNHFRSQHS NNINQFFRSQPF TTLNSFMRSFPP HANLNGWLRTLS SLNNWFRLTPAP HKSPNSLNDFLR ELNTFFRWTTGG TDLNTFLRSLTS NPHLNHIFRSKM IPTLNQHVRASG TLNQWFRPLPTS TLNDFFHNPPHP SLNEHFRPIKQF LITFFRWTTPLK RQNNSLSMFFAG SLNQFFMTSSPA LNQWFRPSPTSG ANSLDNWFRIFP NLNQHIRSLSIP MNLGMDDPRMRR HLLHQPLDGWDL DHGLFTQHIMPD NALDEYFTKPSL 23 Phage display (common panning) 3-4 PMID:19084031 Anti-neuwiedase polyclonal antibodies Zinc metalloproteinase neuwiedase Not determined. Not determined. Ph.D.-12 phage display library (X12) In fact, 80 mimotopes were obtained. However, only 24 mimotopes aligned with the primary structure of neuwiedase were given in the original paper. 572 SKSSITITNKRLTRK 1 Phage display (common panning) 3 PMID:19186111 Anti-SPACc polyclonal antibody IgG Scolex protein antigen from cysticercus cellulosae (SPACc) Not determined. Not determined. G-α phage display library (XXLXXXAX) Four libraries were used in panning against anti-SPACc IgG. Many phage clones were picked and checked. However, only one sequence is given in the original paper. It was showed that SKSSITITNKRLTRK might be devoloped into a promising diagnostics for human neurocysticercosis. 573 SWTWHFPESPPP(9)[+, +, -] SWTFYWPDAQLG(3)[+, +, -] QWQLHWPASKQA(2)[+, +, -] EWTWVFPTTHTS(1)[+, +, -] EWTFQWNSYPAD(1)[+, +, -] EWDFFWPPTQTP(1)[+, +, -] EWQYHWPTLQSR(1)[+, +, -] QWTITYPKPPAL(1)[+, +, -] GWTVFYPDNLRP(1)[+, +, -] QWEWHYMAGYLA(1)[+, +, -] 10 Phage display (common panning) 3 PMID:19465078 Cadherin-1 Cadherin-1 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding of each clone to E-cad/Fc chimeric protein, N-cad/Fc chimeric protein and VE-cad/Fc chimeric protein was confirmed by ELISA, respectively. + denotes phage clone bound to the cadherin ectodomain/Fc chimeric protein and - represents phage clone did not bind to the cadherin ectodomain/Fc chimeric protein. In fact, the target used in the experiment is a chimeric protein composed of the E-cadherin ectodomain fused to the immunoglobulin G1 Fc fragment. The bound phages were eluted through using TBS containing 2 mM EDTA. Total number of clones were 23. All of the phage clones not only bound to the E-cad/Fc chimeric protein, but also to the Ncad/Fc chimeric protein. 574 SWELYYPLRANL(8)[+, +, -][9.4, 0.323] QWEIRYPWPSMG(4)[+, +, -][NT] QWTYYLPLTPRW(2)[+, +, -][NT] EWTYTFPTAHSI(2)[+, +, -][NT] EWFWSWPGYSNT(1)[+, +, -][NT] SWEWYIPYLNRT(1)[+, +, -][NT] AWTWSLPTLPQS(1)[+, +, -][NT] 7 Phage display (common panning) 3 PMID:19465078 Cadherin-1 Cadherin-1 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA,Surface plasmon resonance (SPR) The binding of each clone to E-cad/Fc chimeric protein, N-cad/Fc chimeric protein and VE-cad/Fc chimeric protein was confirmed by ELISA, respectively. + denotes phage clone bound to the cadherin ectodomain/Fc chimeric protein and - represents phage clone did not bind to the cadherin ectodomain/Fc chimeric protein. Besides, the binding of the linear peptide, H-SWELYYPLRANL-NH2 to E-, N-, P- and OB-cad/Fc chimeric proteins was analyzed by SPR. The linear peptide, H-SWELYYPLRANL-NH2 was found to bind to the E- and N-cad/Fc chimeric proteins with high affinities (Kd, μM) of 9.4 μM and 0.323 μM, respectively. NT denotes not tested. In fact, the target used in the experiment is a chimeric protein composed of the E-cadherin ectodomain fused to the immunoglobulin G1 Fc fragment. The bound phages were eluted through using 0.2M glycine HCl, pH 2.2 followed by neutralization with 1 M Tris-HCl, pH 9.1. Total number of clones were 24. All of the phage clones not only bound to the E-cad/Fc chimeric protein, but also to the Ncad/Fc chimeric protein. Peptide SWELYYPLRANL was found to bind both E- and N-cad/Fc chimeric proteins with affinities (KD) of 9.4 μM and 323 nM, respectively, as judged by surface plasmon resonance spectroscopy. 575 CKRDSTWC(8)[1.8 ± 0.12] CKYLWSKC(4)[1.1 ± 0.08] CKYWWSKC(3)[1.3 ± 0.11] CKYWLSRC(3)[1.05 ± 0.13] CKYAWSRC(1)[0.9 ± 0.05] CKYSMSKC(1)[0.8 ± 0.07] K-[RY]-x-W-S-[KR] 6 Phage display (common panning) 4 PMID:19520204 Anti-rabies virus polyclonal antibody IgG Glycoprotein G Not determined. Not determined. fdMED1-based CX6C M13 phage display library ELISA Absorbance measured at 450nm after substraction of background was shown. The selected mimotopes were able to inhibit the interactions of the human anti-rabies virus IgG antibodies with rabies virus in a dose-dependent manner. Subcutaneous administration of phage bearing CKRDSTWC induced an rabies virus glycoprotein-specific IgG response in BALB/c mice. 576 QKTLAKSTYMSA(9) APHWKHKREPPT(1) MPSLNNTESKLG(1) MSQPTTNKMLLS(1) SKKRFPNTSFRQ(1) LPPWKHKTSGVA(1) APPPTHWKKQLY(1) SMEPRKGPKRRG(1) 8 Phage display (common panning) 3 PMID:19100827 Anti-MrkD monoclonal antibody E01 Fimbria adhesin protein Not determined. Not determined. Ph.D.-12 phage display library (X12) This monoclonal antibody was produced by the hybridoma technique using recombinant MrkD-GST as the immunogen. It had an IgG1 isotype and showed high specificity to MrkD protein with an affinity of about 0.3 mg/ml. Thirty-six phage clones were randomly selected and their specificity to mAb was verified by sandwich and competitive inhibition ELISA. Sixteen phage clones were sequenced. 577 GEPQTKLFSFPL(10)[1.13 ± 0.14] 1 Phage display (subtractive panning) 3 PMID:19103307 Anti-CTGF monoclonal antibody 7G2 Connective tissue growth factor Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Microtiter wells were coated with 2 μg/ml of ZD521 peptide, TrxA-ZD521, or TrxA and blocked with PBS containing 10% fetal bovine sera. 7G2(anti-CTGF antibody) in the concentrations of 0, 0.1, 0.5, 1, 2 μg/ml was added to the wells. Bound 7G2 was detected with HRP-conjugated anti-mouse IgG antibody and then with the substrate. The values are recorded at OD450 nm. SDs are indicated by error bars. Only the value of the binding of ZD521 peptide to 7G2 in the concentration of 2 μg/ml was shown. Data were reproduced from the graph. This monoclonal antibody was produced by the hybridoma technique using recombinant CTGF C-terminal domain as the immunogen. The results of ELISA assays and Westen-blot showed that 7G2 could specifically bind to TrxA-CTGF/C and kidney mesangial cell lysates, but not to TrxA. The phage library was reabsorbed by mouse serum IgG and BSA to remove non-specific binders, and then specifically absorbed with 7G2. After 3 rounds of selection, roughly 31.3% (10/32) of the phage clones analyzed exhibited 7G2 binding activity by ELISA. All 10 positive phage clones were sequenced, and the sequences of all clones were identical, i.e. GEPQTKLFSFPL. The antiserum from mice immunized with TrxA-GEPQTKLFSFPL could also bind to CTGF/C recombinant protein and native CTGF, as well as significantly inhibit the proliferation of kidney mesangial cells induced by CTGF/C. 578 SVSVGMLPSPRP(20)[24.07 ± 3.40] SSWILSPYHWGR(13)[34.59 ± 8.20] GSFASLTNPRVL(8)[15.13 ± 1.36] TIQHQNPPHYAV(7)[2.76 ± 0.20] SNPHTDNHWPGR(2)[16.18 ± 1.59] 5 Phage display (common panning) 5 PMID:19389430 Phytophthora capsici extract Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The apparent dissociation constant (Kd, pM) was obtained from a binding saturation curve fitted from three independent experiments. After five rounds of biopanning, 50 plaques were selected and sequenced. All the phages showed high binding affinity for P. capsici in the picomolar range. Phages bearing SVSVGMLPSPRP, TIQHQNPPHYAV and GSFASLTNPRVL inserts showed high specificity toward P. capsici, whereas GSFASLTNPRVL and SNPHTDNHWPGR bearing phages also bound to some of the similar Phytophthora strains with comparable affinities. 579 NHNMHRTTQWPL(2) KVTLHHPPITRS(2) KHLNFLEGRPTF(2) TGLPLYINEGRP(2) YTPQKKIERAFG(1) NPLPSNSPPTRS(1) 6 Phage display (common panning) 3 PMID:19400768 UDP-N-acetylmuramoyl-L-alanyl-D-glutamate--2,6-diaminopimelate ligase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phages with encoded peptides were eluted with 100 μl of 0.2 M glycine/HCl (pH 2.2). NHNMHRTTQWPL inhibited MurE ATPase activity with an IC50 value of 500 μM. The inhibition was shown to be time-dependent and was reversed by the addition of meso-A2pm or UDP-MurNAc-Ala-Glu during the pre-incubation step. MurEp1 was found to interfere in meso-A2pm and UDP-MurNAc-Ala-Glu binding necessary for amide bond formation. 580 KVTLHHPPITRS(2) YTPQKKIERAFG(1) KSASNHQAHWLK(1) TPWHFHSTNGFR(1) INKPFHKVMPYA(1) HSVHSKARHLYT(1) TGLPLYINEGRP(1) YTTSNTLQVIAR(1) SFSNLAPSTRGT(1) 9 Phage display (common panning) 3 PMID:19400768 UDP-N-acetylmuramoyl-L-alanyl-D-glutamate--2,6-diaminopimelate ligase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phages with encoded peptides were eluted with 100 μl of 1mM ATP. NHNMHRTTQWPL inhibited MurE ATPase activity with an IC50 value of 500 μM. The inhibition was shown to be time-dependent and was reversed by the addition of meso-A2pm or UDP-MurNAc-Ala-Glu during the pre-incubation step. MurEp1 was found to interfere in meso-A2pm and UDP-MurNAc-Ala-Glu binding necessary for amide bond formation. 581 NHNMHRTTQWPL(6) YTPQKKIERAFG(2) YPHYSLPGSSTL(1) HWKHEMYPRTRL(1) 4 Phage display (common panning) 3 PMID:19400768 UDP-N-acetylmuramoyl-L-alanyl-D-glutamate--2,6-diaminopimelate ligase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phages with encoded peptides were eluted with 100 μl of 1 mM meso-A2pm. NHNMHRTTQWPL inhibited MurE ATPase activity with an IC50 value of 500 μM. The inhibition was shown to be time-dependent and was reversed by the addition of meso-A2pm or UDP-MurNAc-Ala-Glu during the pre-incubation step. MurEp1 was found to interfere in meso-A2pm and UDP-MurNAc-Ala-Glu binding necessary for amide bond formation. 582 ATLTDLMWFWMG ITIPLYALRSTA 2 Phage display (subtractive panning) 4 PMID:19454218 Anti-GA4 monoclonal antibody 8/E9-Gibberellin A4 complex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Before and after binding reaction with 8/E9 in the presence of GA4, clones that bind 8/E9 Ab in the absence of GA4 were removed with immunotubes that had been coated with 8/E9 Ab in the absence of GA4. The 2 peptides showed specific binding to the complex of the antibody and its ligand GA4; that is, the antibody could not be replaced with the other anti-GA4 antibody, and GA4 could not be replaced with GA1, another ligand of the antibody. ITIPLYALRSTA showed higher GA4 dependency for binding to 8/E9. 583 TMTPPTR(0.066) GNDWPHW(0.066) EHPYITV(0.066) SSLLPTT(0.066) NTNTLHL(0.066) SYPDLHL(0.066) SILPYPY(0.6) 7 Phage display (subtractive panning) 2 PMID:19576904 The idiosyncratic pseudoknot structure (h18) formed by the segment 500–545 of 16S rRNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) he library was initially counter-selected by pre-incubation with the streptavidin-coated solid carrier used in selection. Target-bound phages were released by DNase treatment. A synthetic 59-nt RNA/DNA hybrid that was composed of a 49-nt RNA segment representing h18 of Bacillus anthracis 16S rRNA followed by a 10-nt poly-dA tail with biotin attached at the 3′ end was used as a target for panning. SILPYPY was the only sequence found among 60 sequenced phages after the third round of selection. Such a rapid drop in library complexity was unexpected and might have reflected an unusual susceptibility of the poly-dA tail to DNase I when the RNA/DNA hybrid target was associated with the corresponding phage variant. The peptide SILPYPY moderately stimulated protein synthesis. 584 FPGHSGP(0.02) VQSLPSP(0.02) EPLQLKM(0.02) TPHNTST(0.02) QWTWTQY(0.02) LTHPRWP(0.02) TKTDTWL(0.02) LSPKLPT(0.02) NTPQGMT(0.03) THPLLLS(0.07) GHWEARE(0.07) AVPRASF(0.07) YHPMPVP(0.08) TPTTDGP(0.2) AGAAMSH(0.32) 15 Phage display (subtractive panning) 4 PMID:19576904 The idiosyncratic pseudoknot structure (h18) formed by the segment 500–545 of 16S rRNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The library was initially counter-selected by pre-incubation with the streptavidin-coated solid carrier used in selection. Target-bound phages were released by low-pH elution. A synthetic 59-nt RNA/DNA hybrid that was composed of a 49-nt RNA segment representing h18 of Bacillus anthracis 16S rRNA followed by a 10-nt poly-dA tail with biotin attached at the 3′ end was used as a target for panning. AGAAMSH efficiently interfered with both bacterial and eukaryotic translation. It exhibited a high affinity binding to the isolated small ribosomal subunit (Kd of 1.1 μM). 585 AMSAPIP(94) GTMLAAV(4) MKHPPRI(2) 3 Phage display (subtractive panning) 4 PMID:19576904 The idiosyncratic pseudoknot structure (h18) formed by the segment 500–545 of 16S rRNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The library was initially counter-selected by pre-incubation with the streptavidin-coated solid carrier used in selection. The phage-target complexes were captured in the wells of streptavidin-coated microtiter plate and were eluted with a low-pH buffer. A synthetic 59-nt RNA/DNA hybrid that was composed of a 49-nt RNA segment representing h18 of Bacillus anthracis 16S rRNA followed by a 10-nt poly-dA tail with biotin attached at the 3′ end was used as a target for panning. GTMLAAV and AMSAPIP reduced protein yield by 70-80%. 586 TLTYTWS 1 Phage display (in vivo) 2 PMID:19584266 MMP-2-processed pepsin-extracted collagen IV from human placenta Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In a first step, a pool of potential tumor-homing phage was selected in vivo. In a second step, this phage pool was panned against immobilized MMP-2-processed pepsin-extracted collagen IV from human placenta in vitro. Seven individual phage clones from the second round of in vitro selection were isolated. The phage clone displayed the sequence TLTYTWS could bind to MMP-2-processed collagen IV but not to native collagen IV. 587 LVPPSFS(2) LPLTALP(2) LPLTPLP(2) HPVHHYQ(2) LPGIMSL(1) HKVVAYY(1) HSNTGYP(1) TIGLITS(1) TSGLASR(1) FPLLNML(1) NNLLPPY(1) SIVRLQV(1) SISVIQE(1) QPKQFFQ(1) AQCLRIP(1) 15 Phage display (subtractive panning) 3 PMID:19645415 Hp-AS-pwk RNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Preselection was conducted with prewashed beads to remove streptavidin-binding phage. For screening, the hp-AS-pwkRNA was refolded and hybridized with the 50-biotin-DNA in a 1:1 molar ratio. \r\nThe phages were eluted nonspecifically with 100 μL of buffer B (0.2M glycine-HCl, pH2.2, 1mg/mL BSA) for exactly 9 min followed by neutralization with 15 μL of buffer C (1M Tris-HCl, pH 9.1). HPVHHYQ and LPLTPLP displayed selective binding to the A-site 16S rRNA with on-bead fluorescence assays. 588 LPLTPLP(2) LPLTALA(2) AQATALP(1) LPLTPLA(1) HPVHHYQ(1) TLHPAHP(1) TIGAITS(1) TQSLASR(1) GSWPSLL(1) NWSSLY(1) NAFHSHI(1) SHIMPPN(1) QPTSEGL(1) QCWSPSL(1) QSTLNPT(1) 15 Phage display (subtractive panning) 3 PMID:19645415 Hp-AS-pwk RNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Preselection was conducted with prewashed beads to remove streptavidin-binding phage. For screening, the hp-AS-pwkRNA was refolded and hybridized with the 50-biotin-DNA in a 1:1 molar ratio. The phages were eluted specifically with 100 μL of buffer A containing 300 pmol of nonbiotin-labeled target RNA (hp-AS) for 1 h at room temperature. HPVHHYQ and LPLTPLP displayed selective binding to the A-site 16S rRNA with on-bead fluorescence assays. 589 LPLTPLP(4) SAKLMGH(2) HPVHHYQ(2) LPVTPLP(1) NQDVPLF(1) VSSGPHW(1) TIGAITS(1) 7 Phage display (subtractive panning) 4 PMID:19645415 Hp-AS-pwk RNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Preselection was conducted with prewashed beads to remove streptavidin-binding phage. For screening, the hp-AS-pwkRNA was refolded and hybridized with the 50-biotin-DNA in a 1:1 molar ratio. The phages were eluted nonspecifically with 100 μL of buffer B (0.2M glycine-HCl, pH2.2, 1mg/mL BSA) for exactly 9 min followed by neutralization with 15 μL of buffer C (1M Tris-HCl, pH 9.1). In the fourth round, the competitor RNA (hp-C, which is missing the triple A bulge at nucleotides 1492, 1493, and 1408,) was used in a counter-selection. HPVHHYQ and LPLTPLP displayed selective binding to the A-site 16S rRNA with on-bead fluorescence assays. 590 LPLTPLP(7) LPLTTLH(1) QLPTTLP(1) TIGAITS(1) 4 Phage display (subtractive panning) 4 PMID:19645415 Hp-AS-pwk RNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Preselection was conducted with prewashed beads to remove streptavidin-binding phage. For screening, the hp-AS-pwkRNA was refolded and hybridized with the 50-biotin-DNA in a 1:1 molar ratio. The phages were eluted specifically with 100 μL of buffer A containing 300 pmol of nonbiotin-labeled target RNA (hp-AS) for 1 h at room temperature. In the fourth round, the competitor RNA (hp-C, which is missing the triple A bulge at nucleotides 1492, 1493, and 1408,) was used in a counter-selection. LPLTPLP displayed selective binding to the A-site 16S rRNA with on-bead fluorescence assays. 591 EGEVGLG(0.58) MRRSVGS(0.14) SSAVL(0.08) VLI(0.08) SAGSVAL(0.06) FGVR(0.03) GFWEGGL(0.03) 7 Phage display (in vivo) 4 PMID:19825959 Human breast cancer cell line MDA-MB-231 Not determined. Not determined. Not determined. X9 T7 phage display library Mice bearing MDA-MB-231 human breast tumors were subject to 40 mg/kg sunitinib given i.p. for 3 consecutive d. Treatment began 30 d after tumor implantation. The phage libraries were administered 4 h after the last treatment. Phages were recovered after being in circulation for 16h by harvesting the tumors in the mice. Phages recovered from excised tumors were amplified and subjected to three more rounds of selection. Thirty-six phage plaques were sequenced after the 4th round of panning. EGEVGLG was the dominant sequence isolated from biopanning. This peptide showed increased binding relative to control groups in two cancer cell lines (MDA-MB-435 and MCF-7 human breast) responding to sunitinib treatment, whereas no elevated binding occurred in vitro when samples were incubated with tumor cells that are unresponsive to sunitinib treatment (B16 melanoma and BxPC3 pancreatic). 592 EGEVGLG(0.67) MRRSVGS(0.12) SSAVL(0.18) FGVR(0.01) GFWEGGL(0.03) 5 Phage display (in vivo) 4 PMID:19825959 Human breast cancer cell line MCF-7 Not determined. Not determined. Not determined. X9 T7 phage display library Mice bearing MDA-MB-231 human breast tumors were subject to 40 mg/kg sunitinib given i.p. for 3 consecutive d. Treatment began 30 d after tumor implantation. The phage libraries were administered 4 h after the last treatment. Phages were recovered after being in circulation for 16h by harvesting the tumors in the mice. Phages recovered from excised tumors were amplified and subjected to three more rounds of selection with mice bearing MCF-7 human breast tumors. Thirty-six phage plaques were sequenced after the 4th round of panning. EGEVGLG was the dominant sequence isolated from biopanning. This peptide showed increased binding relative to control groups in two cancer cell lines (MDA-MB-435 and MCF-7 human breast) responding to sunitinib treatment, whereas no elevated binding occurred in vitro when samples were incubated with tumor cells that are unresponsive to sunitinib treatment (B16 melanoma and BxPC3 pancreatic). 593 CGYWRSEWGLC(11)[1.34] CTGYWPKAWGLC(7)[1.37] CTGFWEREWGLC(4)[0.82] CLYWPRLWGLC(1)[0.29] CYWAVRWGLLGC(1)[0.26] CGYWADVWQIHC(1)[0.50] [YF]-W-x(3)-W-G-L 6 Phage display (common panning) 5 PMID:19233852 Human IgG-Fc acid conformer Not determined. Not determined. Not determined. CX7-10C T7 phage display library ELISA The absorbance at 450 nm was measured using a microplate reader. Data were reproduced from the graph. These peptides recognized the non-native conformer (acid conformer) of human IgG that was generated by the low pH treatment, but not the native conformer. 594 CSSAFYPKC(8) CTRQPDRSC(1) CTLQPDRSC(1) CSLQPDRSC(1) 4 Phage display (common panning) 4 PMID:19290051 Anti-BoNT monoclonal antibody F1-40 Botulinum neurotoxin type A, BoNT/A Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 595 IGKIDRV(4) RILASRV(4) IGRRSRV(4) RLRASRV(2) SPRQSRV(2) IGKISRV(2) RLRASWV(2) SRRVSEV(1) AHRVSRI(1) KGRMTRV(1) AGRTTQV(1) SHRQSRV(1) IRRPSIV(1) VIVVSSV(1) TLRESVI(1) WDRASSV(1) PVKFSAV(1) [RK]-x-S-R-V 17 Phage display (common panning) 3 PMID:19235856 PDZ10 domain of Multiple PDZ domain protein 5-hydroxytryptamine receptor 2C Not determined. Not determined. X7 T7 phage display library IGRISRV displayed a two-fold improved affinity over the octapeptide derived from the carboxy terminus of the hc-Kit protein, which was demonstrated as among the highest affinity ligands reported to date for that domain. 596 AWKQTSV(7) FEMGTPV(5) SPRQSRV(3) MVGNMLV(2) EFRESSV(2) AWKQTTV(1) RRVESSV(1) RVRESKV(1) VVIGTSV(1) SEDPIAV(1) WDNGTRV(1) DLRTTSV(1) RDKGTRV(1) RFQETQI(1) 14 Phage display (common panning) 3 PMID:19235856 PDZ3 domain of Postsynaptic density protein 95 (PSD-95) Cysteine-rich PDZ-binding protein Not determined. Not determined. X7 T7 phage display library FEMGTPV possessed a reduced affinity, which was about 10-fold lower than that of benchmark KKETEV peptide, but comparable with the smallest recognizable CRIPT peptide fragment (KQTSV) binding. 597 CQQSNRGDRKRC(3) CMGNKRSAKRPC(3) CESHRQRRAKC(2) CKRTSKCGGKC(1) CLRKRRENTKC(1) CHHWTFRKTTC(1) CSPNNTRRPNK(1) 7 Phage display (common panning) 5 PMID:19123480 Embryonal Rhabdomyosarcoma cell line RD Not determined. Not determined. Not determined. CX7-10C T7 phage display library Sixteen clones were sequenced. CQQSNRGDRKRC and CMGNKRSAKRPC binded to rhabdomyosarcoma and to other tumour cell lines, but not to normal skeletal muscle cells and fibroblasts. 598 RCMTSRS(9) LATTVPH(4) TATTIPT(1) 3 Phage display (common panning) 3 PMID:19811729 Ouabain Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) RCMTSRS could reverse the growth inhibition and death induction of ouabain in EAhy926 cells. 599 TPMNHHSQHAER(16)[1.12 ± 0.02] AHLPIVRASLPS(12)[1.33 ± 0.07] FPSSLIIPPLPN(7)[0.32 ± 0.06] GNIIPDRPMHPT(5)[0.64 ± 0.03] 4 Phage display (subtractive panning) 4 PMID:19550037 Disintegrin domain of ADAM 15 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Binding was detected using the anti-M13 antibody and expressed as the absorbance (405 nm). Results are expressed as the mean ± SD of triplicate experiments. Data were reproduced from the graph. Two rounds of subtractive selection with streptavidin were performed. AHLPIVRASLPS was found to be homologous with integrin 600 VRKRSECLGAHD(19) 1 Phage display (subtractive panning) 3 PMID:19778796 Hepatocellular carcinoma cell line HepG2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Subtractive selection with human liver cell line L02 cells was performed. Among 30 clones picked, 27 clones have proper insetions. However, only the most frequent sequence is given in the original paper. Immunocytochemistry and immunohistochemistry confirmed the specificity of the VRKRSECLGAHD bearing phage binding to the hepatoma cells. 601 CIATPYKGC(2) CDDRPPKSC(1) CRPSTAPMC(1) CSHSQHEMC(1) CSSHKNMAC(1) CNHNHRDPC(1) CHGVLPLSC(1) CSPLQAPYC(1) CSSRSLPDC(1) CGSLALNLC(1) CDHRGHYGC(1) CPPNSTPVC(1) CHWSCPPCC(1) CYPRNVLGC(1) CPLRPNAQC(1) CHLFSPLPC(1) CWSERSQKC(1) CHLHNYRLC(1) CNTAHGLLC(1) CPTPSPALC(1) CATGSFPKC(1) CQSNTTQEC(1) CQALYKQLC(1) CARNAESMC(1) CLIGVPRLC(1) CLQRMSYMC(1) CVARPLPHC(1) CNPTRAQWC(1) CPFLKGHTC(1) CPGARNAQC(1) CFAYGSSFC(1) CQPTFQSRC(1) CALRGHPHC(1) CHKSYMLKC(1) CSSLFWHRC(1) 35 Phage display (common panning) 3 PMID:19275745 Protein SMG7 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) CDDRPPKSC can bind specifically to SMG7. 602 SHWWWWDARGYD(5) WHYPRWYYPPYN(2) FHWSWYTPSRPS(1) WHWRNPDFWYLK(1) SVSVGMKPSPRP(1) 5 Phage display (common panning) 4 PMID:19778804 Natriuretic peptides B Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 603 MAHRHPQ(2) AFHGPVH(2) GTTVIAR(1) YQDSAKT(1) DWPLQKL(1) NSFAPQT(1) KTGHTSS(1) KGHSLMP(1) TIPSPKP(1) KVSMNSS(1) NRLGSTG(1) LVQPHTR(1) STEIELY(1) QHSEGAL(1) NTPNLWN(1) STSVLYN(1) SYNQAYP(1) NNVQLPN(1) THDHLPN(1) IPTLPSS(1) QLPGRSG(1) HAIYPRH(1) HYRFDLT(1) LPLTPLP(1) 24 Phage display (common panning) 4 PMID:19275099 Anti-Human KGF monoclonal antibody Keratinocyte growth factor, KGF Not determined. Not determined. Ph.D.-7 phage display library (X7) The monclonal antibody was bought from R&D Systems. However the clone number of the antibody is not given in the original paper. There are two mouse anti-human KGF mabs from R&D Systems: Clone 29522 and Clone 29568. Both of them belong to IgG1. Clone 29522 can be used in ELISA Capture (Matched Pair)and neutralization; Clone 29568 is for western blot. Thus, we guess the antibody used in this study is from clone 29522. 604 CEPYYPSYC(11) CTSYYPSYC(1) YYPSY 2 Phage display (common panning) 4 PMID:19857112 Anti-BoNT monoclonal antibody F1-2 Botulinum neurotoxin A heavy chain Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Twelve clonal plaques were picked for DNA sequencing. 605 CQAYYPSYC(6) CPTWYPKSC(1) YYPSY 2 Phage display (common panning) 4 PMID:19857112 Anti-BoNT monoclonal antibody F1-5 Botulinum neurotoxin A heavy chain Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Twelve clonal phage plaques were sequenced for each antibody, with five plaques from the F1-5 experiment failing to yield correct sequences. 606 SYNFNWEYKRHE GLKQSFSWERFK TEALHYTWEHKK SFMFPWEMRPPL NPQAYVWEKKLR 5 Phage display (common panning) 5 PMID:19951177 Anti-RBD of SARS-CoV spike protein monoclonal antibody S1 Spike glycoprotein Not determined. Not determined. M13LP67-based X12 phage display library 607 TMWPKSDDMRSL 1 Phage display (common panning) 5 PMID:19951177 Anti-RBD of SARS-CoV spike protein monoclonal antibody S2 Spike glycoprotein Not determined. Not determined. M13LP67-based X12 phage display library 608 YTMKGDTARATA EPSPLKGDVIRT NVYIAKGDSTRK THLIVPKGDHIR 4 Phage display (common panning) 5 PMID:19951177 Anti-RBD of SARS-CoV spike protein monoclonal antibody S3 Spike glycoprotein Not determined. Not determined. M13LP67-based X12 phage display library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hage display (common panning) 3-4 PMID:18701808 Bone-like mineral (BLM) films; Hydroxyapatite (HA) disks Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA To determine the binding efficiency of the identified phage to the bone-like substrate materials, an ELISA was performed on BLM films, HA disks, PLGA films, and tissue culture polystyrene (TCPS) wells. In the blocked plate, dilutions of individual phage were prepared in TBST (0.1%) in the concentrations of 1.0e6, 1.0e7, 1.0e8, and 1.0e10 pfu (n = 1 per dilution). HRP-conjugated anti-M13 antibody was prepared in 1:5000 ratio in blocking buffer and added to each well. An aliquot of each well was read on an UV Spectrophotometer (Biorad SmartSpec 3000) at 410 nm. OD410 values of the binding of the identified phage in the concentration of 1.0e10 pfu to HA, BLM, TCPS and PLGA were shown respectively and data were reproduced from the graph. NT denotes not tested. The DNA from a total of 243 clones was sequenced. Nineteen phage sequences emerged multiple times after DNA sequencing (Top19 sequences). Of the 19 clones, 17 were found in more than 1 experiment on the same substrate, either BLM or HA. Of these 17 phage clones, 7 had high affinity towards both BLM and HA (Top7 sequences). Ten of the identified phage were analyzed using a modified ELISA on BLM films, HA disks, PLGA films and tissue culture polystyrene (TCPS) wells at 4 phage dilutions. Low adsorption of the selected phage to TCPS and PLGA compared to HA and BLM illustrates the identified phage have specificity to apatite-based substrates. Of the 10 peptides tested, favorable phage binding was observed for all sequences except SSLGLTVSSIMY, VSPLSFGSPRYP and WSPAPHVIMGTT. On the other hand, the peptides having the highest affinity and greatest potential for adsorption on HA disks and BLM films include APWHLSSQYSRT, STLPIPHEFSRE and VTKHLNQISQSY. The peptide VTKHLNQISQSY is similar in composition to regions found in fibromodulin, lumican and decorin. Overall, STLPIPHEFSRE and VTKHLNQISQSY have a significantly higher adsorption to the apatite-based materials in comparison to APWHLSSQYSRT. Pay attention to STLPIPHEFSRE. This sequence occurred only once in the 3 phage display experiments; however, it was also included into the "multiple times" group by the authors. 610 CTTQGNPQC(1) CATTKFSGC(1) CLSEQNRSC(1) CKLGFHGKC(1) CKTHAQHEC(1) CYEQHHPGC(1) CTTPHAWLC(1) CPMPRPSSC(1) CTDNTAKNC(1) CSLTTSTLC(1) 10 Phage display (in vivo) PMID:19007831 Rat nose-to-brain Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The C7C phage peptide display library (Ph.D.-C7C) was applied intranasally to anesthetized rats and recovered phage from the brain tissue 45 min after phage administration. After three rounds of panning, 10 positive phage clones were selected and sequenced. After nasal administration, Clone7 entered the brain within 30 min and exhibited translocation efficiency about 50-fold higher than a random phage. A 11-amino acid synthetic peptide derived from the displayed sequence of Clone7 (ACTTPHAWLCG, the flanking A and G were derived from the M13 coat protein) efficiently inhibited the nasal-brain translocation of Clone7. Both phage recovery results and fluorescent microscopy images revealed the presence of many more Clone7 phage in the brain than in the liver, kidney and other internal organs after the nasal administration, suggesting that Clone7 bypassed the BBB and entered brain directly. Furthermore, both Clone7 and the ACTTPHAWLCG peptide were found to be heavily distributed along the olfactory nerve after the nasal administration, further suggesting a direct passage route into the brain via the olfactory region. 611 FHRWPTWPLPSP(8) MHRHPPPITLPL(3) FPWHFFSPQLRG(3) MHRHPPPITTAA(3) AAFHRAHHLTSP(2) FHSNWPKGSTSL(2) FHRWPTWPTSFS(2) MHRPHFNPTLAT(2) F-H-R-H-P(2)-x-P-T-L-x(2) 8 Phage display (common panning) 3-4 PMID:19115038 Monocyte differentiation antigen CD14 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) For the first and second rounds, bound phages were eluted by adding 1 ml of elution buffer (0.1 N HCl adjusted to pH 2.2 with glycine, 1 mg/ml BSA, and 0.1 mg/ml phenol red). The bound phages in the third and fourth rounds were eluted by adding 1.5 ml of lipopolysaccharide-binding protein (10 μg/ml, R&D Biolabs). One hundred clones were randomly singled out. Twenty-five phage clones showing higher rhCD14 binding activity were sequenced. FHRWPTWPLPSP can markedly inhibit LPS induced TNF-alpha expression. 612 GLHVMHKVAPPR 1 Phage display (common panning) 5 PMID:19117819 Zinc oxide, ZnO Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) GLHVMHKVAPPR bound to and directed the growth of ZnO hexagonal nanocrystals. By altering the concentration of Z1 peptide, the ZnO nanocrystal morphology can be tailored. Additionally, GLHVMHKVAPPR was used to direct the growth of ZnO structures on free-standing silk films. The results demonstrated the utility of peptides in controlling the structure and deposition of ZnO. 613 QQMHLMSYAPGP(19) AHRHPISFLSTL(12) VEAPLHRAQPHY(10) KMDRHDPSPALL(8) AYYPQNHKSKAE(2) APNHIPRPPGLT(1) YPHYSLPGSSTL(1) 7 Phage display (subtractive panning) 3 PMID:21475935 NCI-H1299 non-small cell lung cancer cell line Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) NCI-H1299 cells were selected as the target cells, and the normal lung small airway epithelial cells (SAE cell line) were used as the absorber cells for a whole-cell subtractive screening. After in vitro selection, 60 individual clones were chosen and sequenced. Seven of these clones were found to be lacking exogenous sequences. QQMHLMSYAPGP-bearing phages binded most effectively to NCI-H1299 cells. The synthetic peptide QQMHLMSYAPGP was demonstrated to be capable of binding to the tumor cell surfaces of NCI-H1299 and A549 cells and biopsy specimens, but not to normal lung tissue samples, other cancer cells, or non-tumor adjacent lung tissues. 614 CNTPLTSRC(1.0) 1 Phage display (competitive panning) 1 PMID:19447041 Concanavalin-A, Con A Methyl α-D-mannopyranoside Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool The bound phages were eluted by addition of glucose solution (500 mM glucose in TBST). CNTPLTSRC bound strongly to ConA, LCA, and PSA. In addition, it showed marked selectivity by exhibiting weaker affinity toward three mannose-only specific lectins (GNA, HHA, and NPA). 615 STLHALDSHLAL(0.36) 1 Phage display (competitive panning) 2 PMID:19447041 ConA and Lentil lectin Methyl α-D-mannopyranoside Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool In the first round, the target was ConA; in the second round, the targets were ConA and LCA. The bound phages were eluted by addition of glucose solution (500 mM glucose in TBST). STLHALDSHLAL showed a similar binding profile to that of the negative control phage in ELISA. 616 HWDPFSLSAYFP(0.18) 1 Phage display (competitive panning) 3 PMID:19447041 ConA, LCA and Pea lectin Methyl α-D-mannopyranoside Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool In the first round, the target was ConA; in the second round, the targets were ConA and LCA; in the third round, the targets were LCA and PSA. The bound phages were eluted by addition of glucose solution (500 mM glucose in TBST). However, no binding experiment was done with HWDPFSLSAYFP. 617 CKPHASSMC(1.0) 1 Phage display (competitive panning) 1 PMID:19447041 ConA and Lentil lectin Methyl α-D-mannopyranoside Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool The bound phages were eluted by addition of glucose solution (500 mM glucose in TBST). CKPHASSMC bound strongly to ConA, LCA, and PSA. In addition, it showed marked selectivity by exhibiting weaker affinity toward three mannose-only specific lectins (GNA, HHA, and NPA). 618 CSRILTAAC(1.0) 1 Phage display (competitive panning) 1 PMID:19447041 LCA and Pea lectin Methyl α-D-mannopyranoside Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool The bound phages were eluted by addition of glucose solution (500 mM glucose in TBST). CSRILTAAC bound strongly to ConA, LCA, and PSA. In addition, it showed marked selectivity by exhibiting weaker affinity toward three mannose-only specific lectins (GNA, HHA, and NPA). 619 CSPIYKDTC(1.0) 1 Phage display (competitive panning) 1 PMID:19447041 ConA and Pea lectin Methyl α-D-mannopyranoside Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool The bound phages were eluted by addition of glucose solution (500 mM glucose in TBST). CSPIYKDTC bound strongly to ConA, LCA, and PSA. In addition, it showed marked selectivity by exhibiting weaker affinity toward three mannose-only specific lectins (GNA, HHA, and NPA). 620 YQTSSPAKQSVG(1.0) 1 Phage display (competitive panning) 2 PMID:19447041 ConA, LCA and Pea lectin Methyl α-D-mannopyranoside Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool In the first round, the targets were ConA and PSA; in the second round, the targets were PSA and LCA. The bound phages were eluted by addition of glucose solution (500 mM glucose in TBST). YQTSSPAKQSVG bound strongly to ConA, LCA, and PSA. In addition, it showed marked selectivity by exhibiting weaker affinity toward three mannose-only specific lectins (GNA, HHA, and NPA). 621 CKQMGTLKC(1.0) 1 Phage display (competitive panning) 1 PMID:19447041 ConA and Pea lectin Methyl α-D-mannopyranoside Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool The bound phages were eluted by addition of glucose solution (500 mM glucose in TBST). CKQMGTLKC bound strongly to ConA, LCA, and PSA. In addition, it showed marked selectivity by exhibiting weaker affinity toward three mannose-only specific lectins (GNA, HHA, and NPA). 622 ADPQFSGHTPPQ(0.75) VPPTRPATSTQL(0.25) 2 Phage display (competitive panning) 2 PMID:19447041 ConA, LCA and Pea lectin Methyl α-D-mannopyranoside Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool In the first round, the targets were PSA and ConA; in the second round, the targets were ConA and LCA. The bound phages were eluted by addition of glucose solution (500 mM glucose in TBST). ADPQFSGHTPPQ bound to ConA, LCA, and PSA and showed marked selectivity by exhibiting weaker affinity toward three mannose-only specific lectins (GNA, HHA, and NPA). However, no binding experiment was done with VPPTRPATSTQL. 623 CNNPRAINC(1.0) 1 Phage display (competitive panning) 1 PMID:19447041 Pea lectin and Lentil lectin Methyl α-D-mannopyranoside Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool The bound phages were eluted by addition of glucose solution (500 mM glucose in TBST). Initial assay optimization studies suggested that the majority of the clones from the C-X7-C library demonstrated higher specificity than the X12 library. An exception to this trend was CNNPRAINC, which therefore was dropped from further study. 624 CKSSLLRNC(4) CQDSLLPSC(1) CYTNLKSMC(1) CALPSSTLC(1) CLASSHANC(1) CPSLSSPFC(1) CTSALQMTC(1) CSPFASLMC(1) CTPRAPLLC(1) CYTPKATRC(1) CTRTSPPHC(1) CNARATHNC(1) CTANNAKAC(1) CYPKMNAFC(1) CLDRHPKYC(1) CWADKIQSC(1) CQADKHNKC(1) CSLHHNKIC(1) CHSTPSAHC(1) CHHPQQRQC(1) CHPTVVYGC(1) CHTQISRSC(1) CINSQTIQC(1) CISSIPHQC(1) CLSPRPAMC(1) CPQAGSRDC(1) CQPPIGRIC(1) CQTQSHRFC(1) CSHQPNTNC(1) CTHKLYKNC(1) [LP]-K-S(2)-L(2), NAKATR, ADKHNK, T-P-x-A 30 Phage display (common panning) 3 PMID:19632196 Anti-NAV polyclonal antibody IgG Taiwan cobra venom Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) After three rounds of biopanning, 33 phage clones were randomly isolated and sequenced. Among them, 30 different sequences were obtained. The KSSLLRN-carrying phage occurred four times in selected clones and showed the strongest reactivity to NAV monovalent antivenin.3 625 ILLPQPPKLLLP(2) QLTVPTPQPPKP(2) HFGLGILPQPPK(2) APLPQPNKPHLF(2) IVKPVPPKPRT(2) SVEPQPPKADFH(1) TVLPQPSKALPL(1) LPNPYPQPSKVL(1) WSDPQPQKPAQR(1) SILPQPAKPNSE(1) QVLPHPTKAEHF(1) LVLPHPTKMIES(1) AMPIPHPPKPRT(1) HNWYTHLLPPK(1) NLSHPMLSKPWV(1) SYIPVTPHANGL(1) INTWNYPTANPH(1) SWQESLNVPAVL(1) VNNWESRSPPPG(1) HNNWSTHVVTLT(1) GLSALIADLSP(1) P-[QH]-P-x-K 21 Phage display (common panning) 3 PMID:19772881 Anti-enteroaggregative Escherichia coli (EAEC) polyclonal antibody IgG Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Twenty-six clones were picked and sequenced after panning against IgG purified from sera of a child patient naturally infected with enteroaggregative Escherichia coli (EAEC). The binding reactivity of phage clone bearing NLSHPMLSKPWV was not determined. The binding reactivities of SYIPVTPHANGL, INTWNYPTANPH, SWQESLNVPAVL, VNNWESRSPPPG, HNNWSTHVVTLT and GLSALIADLSP to IgG used for library screening were as weak as control. 626 TYPGYINHSKAS(2) TIPGYINWRGGP(2) TFFPSTPGPRYT(2) HHTWPGYINYV(1) QGIHPYPGYINH(1) QAPGWINVPPIE(1) NHQYYQVPGSIN(1) THHTSWPGQINS(1) SHHQNMVPGSLN(1) EDYRHSRPGRLN(1) QFPGFYNSSPPY(1) SHTTGVPGSVNH(1) MLPGWYNSLQSR(1) HLNKDANLSFAW(1) YLRAPPQWLTNT(1) P-G-x-[ILYV]-N 15 Phage display (common panning) 3 PMID:19772881 Anti-serine protease pet autotransporter polyclonal antibody IgG Serine protease pet autotransporter Not determined. Not determined. Ph.D.-12 phage display library (X12) The binding reactivities of SHHQNMVPGSLN, EDYRHSRPGRLN and YLRAPPQWLTNT to IgG used for library screening were as weak as control. 627 CTLDDSSTC(2) CSPDDSSTC(1) CAPDDSSTC(1) CLADDSSTC(1) CIDDSSMSC(1) CAHRDDSSC(1) CTYALLSSC(1) CVPNYKNQC(1) D(2)-S(2) 8 Phage display (common panning) 2 PMID:19772881 Anti-serine protease pic autotransporter polyclonal antibody IgG Serine protease pic autotransporter Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The binding reactivities of these phage clones were not determined. 628 CSPDDSSTC(3) CLLDDSSTC(2) CNPDDSSMC(1) CSPDDSSMC(1) CSPDDSSAC(1) CVPDDSSTC(1) CTLDDSSTC(1) CSVDDSSLC(1) [PLV]-D(2)-S(2) 8 Phage display (common panning) 3 PMID:19772881 Anti-serine protease pic autotransporter polyclonal antibody IgG Serine protease pic autotransporter Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The binding reactivity of phage clone bearing CSPDDSSAC was not determined. 629 FDYSKHV(17) AYDTEYV(3) 2 Phage display (common panning) 3 PMID:19811418 Anti-PfMIF monoclonal antibody B9 Macrophage migration inhibitory factor-like protein Not determined. Not determined. Ph.D.-7 phage display library (X7) 630 ESAHHHHWKNFR(11) HWWNTPWAWSHP(6) SPQQLKPWWHSP(3) 3 Phage display (common panning) 3 PMID:19811418 Anti-PfMIF monoclonal antibody B9 Macrophage migration inhibitory factor-like protein Not determined. Not determined. Ph.D.-12 phage display library (X12) 631 HNYESPLLPTSH(2) AEHMIWMSPGRA(1) SHHHRDQMDPKH(1) TMGTTSPQIHRY(1) VSEQHQRYIWSS(1) DAVRPITSSAGN(1) ALSSNHPPAQPT(1) NTRLNNTPDSQL(1) TVMMNKEMLTTR(1) 9 Phage display (competitive panning) 2 PMID:19846220 Anti-lipid-transfer protein polyclonal antibody IgE Non-specific lipid-transfer protein, LTP Not determined. Not determined. Ph.D.-12 phage display library (X12) The target was immobilized on beads via a monoclonal mouse-anti-human IgE. In the second round, binding clones were eluted with Tri a 14. 632 SPHDIKRWTMPH(1) NTSHPPYTHRTM(1) AQKPSLGHLSLT(1) DICTNQCARPTG(1) TATNLLTPRWIT(1) EFTEPHSSQLRD(1) HNKGSPLLPTSH(1) KRTPSGIEPALH(1) NHSFHIIATPAP(1) SFKTHNTWYKLG(1) 10 Phage display (competitive panning) 2 PMID:19846220 Anti-lipid-transfer protein polyclonal antibody IgE Non-specific lipid-transfer protein 1, LTP 1 Not determined. Not determined. Ph.D.-12 phage display library (X12) The target was immobilized on beads via a monoclonal mouse-anti-human IgE. In the second round, binding clones were firstly eluted with Tri a 14 and the non-eluted phages were then incubated with Pru p 3, eluting Pru p 3-specific clones. 633 VWALKSHSHKDI(2) KPEFTRTTHNAP(1) NKNPNMIAQFPN(1) GVSVTSISNIAP(1) HNKESPLLPTSH(1) DNRMFRRLSLRV(1) HNYESPRLPTSH(1) HNYESPLLPTRR(1) HNYESPLLPTSH(1) 9 Phage display (competitive panning) 2 PMID:19846220 Anti-lipid-transfer protein polyclonal antibody IgE Non-specific lipid-transfer protein 1, LTP 1 Not determined. Not determined. Ph.D.-12 phage display library (X12) The target was immobilized on beads via a monoclonal mouse-anti-human IgE. In the second round, binding clones were eluted with Pru p 3. 634 GQSALLSMRHKP(1) ALSLPQTTRALF(1) NDLALQPRNAGP(1) IPLSEFLFADPG(1) HNYESPLLPTSH(1) VMAQTNLFSPRS(1) HNYESPLLLRRM(1) FTTLQPPLLPHQ(1) HQAPQPKPNRLF(1) FTTLQPPLLPXQ(1) 10 Phage display (competitive panning) 2 PMID:19846220 Anti-lipid-transfer protein polyclonal antibody IgE Non-specific lipid-transfer protein, LTP Not determined. Not determined. Ph.D.-12 phage display library (X12) The target was immobilized on beads via a monoclonal mouse-anti-human IgE. In the second round, binding clones were firstly eluted with Pru p 3 and the non-eluted phages were then incubated with Tri a 14, eluting Tri a 14-specific clones. 635 VYPTLHFSARGSG QSQWPVLFFSARG SNIPFQFSARGPG SENLQFSARGPGG HMEGSELSFSARG AHQDETLAFSARG NHPFNALSFSSRG YADLLAQFSSKSG YLPSQISAIGRAG NPVIRYASRSHSG FSARG 10 Phage display (common panning) 3 PMID:20090838 Serine-aspartate repeat-containing protein G Fibrinogen beta chain Not determined. Not determined. Ph.D.-12 phage display library (X12) In fact, the target used in this study was N2 and N3 domain of SdrG (SdrGN2N3). 636 HWRTHHIHHFHQG MSPHHMHHSHGHG AKLAHHIHHFHGG WVPHHIHHFHRAG YTHHHHSWRLHTG AHHPHAWRHSHKG STFHFHTHKARHG HSQHHRFHHTYPG H(2)-[IM]-H(2)-F-H 8 Phage display (common panning) 3 PMID:20090838 Serine-aspartate repeat-containing protein C Neurexin-1-beta Not determined. Not determined. Ph.D.-12 phage display library (X12) In fact, the target used in this study was N2 and N3 domain of SdrC (SdrCN2N3). Fifty randomly picked plaques were selected and the corresponding phages were screened again for binding to SdrCN2N3, to SdrGN2N3 and to BSA as controls. All clones bound to SdrC, however, phage from eight clones displayed significantly higher binding to SdrCN2N3 compared to SdrGN2N3 and BSA. The clone inserts were sequenced. 637 VSGKAGYYSRMH SLTPNSKTRLYA HNPQFNLWHRDT TLAKMGLSPLHG HSSWWLALAKPT HNWHWKHKWMPR HYPWFKARLYPL HYPWSKYPTRSP WHDRLYLKWPHR HLGLYNLLRPWP TSNPHTRHYYPI HMRLSQWPLLKP HYPW, KARLY 12 Phage display (common panning) 3 PMID:20082398 GTPase HRas Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In fact, the target used in the panning was immobilized GST-H-Ras G12R/A59T-GMPPNP. HYPWFKARLYPL binds Ras in both the active GTP- and inactive GDP-bound conformations with low micromolar dissociation constants. The dodecamer does not alter the intrinsic GTPase activity of Ras, does not compete for Ras binding to the Ras binding domain of Raf, and does not alter cell viability. 638 HSWYPVPIPPSS(12) SVSPTWPLPPFP(2) HLWHLPPRTPTA(1) APGDMLPPLPSL(1) SVSVGMKPSPRK(1) P-x(2)-P 5 Phage display (competitive panning) 5 PMID:20024074 Tyrosine-protein kinase Lck Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In fact, the target used in the panning was immobilized GST-tagged LckSH3. The sequences of the phage-displayed peptides obtained after five rounds of selection against GST-tagged LckSH3 with the competing GST-tagged SH3 domains from Hck, Src, Pi3k and Abl present in the washing buffer. 639 HPNVLKAPSSNT AETVESCLAKSH VSVWSYQYWRFP KGWMAYPVASSW SSATMTQDSRLI TARNETPAMRMN THPVTDPIAHPP MLPGTSGVTDFQ TFRSNDSYFHQK NLYSNRSAVTSL ASPASHGNNNIS STTVRPHTSDVW 12 Phage display (competitive panning) 2 PMID:20205701 IgE of melon-allergic patients Profilin Not determined. Not determined. Ph.D.-12 phage display library (X12) The target was immobilized on beads via a monoclonal mouse-anti-human IgE. In the second round, binding clones were eluted with recombinant Cuc m 2. All clones were recognized by the serum pool of melon patients in ELISA assays. By contrast, a pool of sera from nonfruit atopic patients recognized none of the selected clones. The peptides included in the selected clones could bind to the specific IgE of melonallergic patients. 640 PFARAPVEHHDVVGL(2) GLDLLGDVRIPVVRR(2) RFLEFPRFFPAIILP(1) PGRLLPGVIQRHFFI(1) GAFSSPRSLTVPLRR(1) CGGLFAGCAALIDVF(1) EFPTFSWSYINDSLL(1) ADWPHARGKFALGNA(1) GFTDVHLHLPGNSHR(1) PVAGMPLFPTAWFAH(1) SAYAATVRGPLSSAS(1) RYASQLSDQILFTLP(1) RDGAFSPPVRWWSFS(1) TGAQAGLHEWRPWGV(1) EDWFSASIRRVPTFA(1) GTWSSTCPLCSATAV(1) PWLPSNLGSRPGLMR(1) 17 Phage display (common panning) 3 PMID:20162339 Brain (brain endothelium) Not determined. Not determined. Not determined. f3-15mer phage display library (X15) In fact, phages were infused via the heart. The major advantage compared to direct infusion into the brain is that the phage have to travel some distance before they reach the brain. On their way, they already encounter endothelial cells: the endothelium of the aorta and carotid arteries. Phage with affinity to ubiquitous endothelial receptors are given the opportunity to bind to these cells and will not reach the brain. This negative selection filters out unspecific binders and allows for the identification of peptides that bind more specifically to brain endothelium. These sequences were deduced from clones isolated from anterior cerebrum. 641 GLDLLGDVRIPVVRR(2) PFARAPVEHHDVVGL(2) GVVNYARAFNVGAAV(1) HAAFEPRGDVRHTLL(1) GDGRFHFLRGFFDSD(1) VRSIALFPPEWSATS(1) GLAHSFSDFARDFVA(1) VTGTQIRLPAYLRFD(1) GYRPVHNIRGHWAPG(1) ITRGGYVIYHDALLA(1) GAYFLSNHAVVRGVG(1) VGRPGGLVGGFASSL(1) LGRAGQSYPSFARGL(1) VVSSRSVLSSQYRGH(1) ALPCNGAGCSRVTAR(1) VPMGLGFLGRGLAPL(1) RSSHHPSFAVSLEPL(1) 17 Phage display (common panning) 3 PMID:20162339 Brain (brain endothelium) Not determined. Not determined. Not determined. f3-15mer phage display library (X15) In fact, phages were infused via the heart. The major advantage compared to direct infusion into the brain is that the phage have to travel some distance before they reach the brain. On their way, they already encounter endothelial cells: the endothelium of the aorta and carotid arteries. Phage with affinity to ubiquitous endothelial receptors are given the opportunity to bind to these cells and will not reach the brain. This negative selection filters out unspecific binders and allows for the identification of peptides that bind more specifically to brain endothelium. These sequences were deduced from clones isolated from posterior cerebrum. Two phage clones, displaying GLAHSFSDFARDFVA and GYRPVHNIRGHWAPG 15 amino acid-peptides (GLA and GYR) that were selected for brain binding in the mouse model, showed significant binding to human brain endothelium (hCMEC/D3), compared to a random control phage. This binding was not seen for other human endothelial cells (HUVEC). Binding to hCMEC/D3 cells was dose dependent. When phage GLA and GYR were individually perfused through the murine brain, their ability to bind to the brain was 6-fold (GLA) and 5-fold (GYR) higher than the control phage. When compared to lung perfusion, phage showed an 8.5-fold (GYR) and 48-fold (GLA) preference for brain over lung compared to the control. 642 GVDQHDNGSRAR(2) WPLHDGGAVNRR(2) FTQLNPTDGGSA(1) TNSFNDSGSAPH(1) H-D-G(2)-S-x-A 4 Phage display (common panning) 3 PMID:19261354 Anti-Toxoplasma gondii tachyzoites monoclonal antibody A4D12 SAG2 protein Not determined. Not determined. Ph.D.-12 phage display library (X12) 643 VQGMLRQDTQNL(1) TSDRDHQNLLAY(1) YNIHEHQNLRAV(1) IIQERQNIWTAP(1) ERSLWQEHQNFP(1) NNWDYPQERQNI(1) AQEHQNLTWRPI(1) DRQTWQNFMLIP(1) QEHQNL 8 Phage display (common panning) 3 PMID:19819255 Anti-chCD132 monoclonal antibody C10 Cytokine receptor common subunit gamma (CD132) Not determined. Not determined. Ph.D.-12 phage display library (X12) 644 KIELPSL(10) KIELPSL 1 Phage display (common panning) 3-4 PMID:19923010 Anti-chicken IL-2 monoclonal antibody 7G5 Interleukin 2 Not determined. Not determined. Ph.D.-7 phage display library (X7) 645 EHLDWFEDWALW(5) FHQQNYNRSIYL(2) EHLDFLDSENTT(1) TYDNDMLYKNHV(1) TGNEKWLYGMLT(1) E-H-L-D-x-N-D-S-L-Y-L 5 Phage display (common panning) 3-4 PMID:19923010 Anti-chicken IL-2 monoclonal antibody 6G5 Interleukin 2 Not determined. Not determined. Ph.D.-12 phage display library (X12) 646 NHLNGSY(8) SILPYPY(1) NHKKGQQ(1) N-H-L-x-G-x-Y 3 Phage display (common panning) 3-4 PMID:19923010 Anti-chicken IL-2 monoclonal antibody 2F9 Interleukin 2 Not determined. Not determined. Ph.D.-7 phage display library (X7) 647 TYWSPTP(4) WHVPFSL(1) TWAWRTP(1) WHPLLSP(1) WHLPRVI(1) WHLPWLE(1) RNVPPML(1) W-H-L-P(2)-SL 7 Phage display (common panning) 3-4 PMID:19923010 Anti-chicken IL-2 monoclonal antibody 7B7 Interleukin 2 Not determined. Not determined. Ph.D.-7 phage display library (X7) 648 YVTKIAS(5) EFNAARL(2) EFNPSNL(2) FKASLLH(1) E-F-K-A-S-x-L 4 Phage display (common panning) 3-4 PMID:19923010 Anti-chicken IL-2 monoclonal antibody 2G11 Interleukin 2 Not determined. Not determined. Ph.D.-7 phage display library (X7) 649 TENPFPE(3) QFNPFPE(2) WENPYPE(2) YYNPYPE(1) TYNPFPE(1) AFNPFPE(1) TENPFPE 6 Phage display (common panning) 3-4 PMID:19923010 Anti-chicken IL-2 monoclonal antibody 3E7 Interleukin 2 Not determined. Not determined. Ph.D.-7 phage display library (X7) 650 PKISGLYADLLAK(7) PGVYINTNRETSS(2) PGPTGIDLWGAEV(1) SGLYL 3 Phage display (common panning) 3-4 PMID:19923010 Anti-chicken IL-2 monoclonal antibody 8H8 Interleukin 2 Not determined. Not determined. Ph.D.-12 phage display library (X12) 651 AHGYWEL(10) AHGYWEL 1 Phage display (common panning) 3-4 PMID:19923010 Anti-chicken IL-2 monoclonal antibody 4F3 Interleukin 2 Not determined. Not determined. Ph.D.-7 phage display library (X7) 652 HHGYWEL(10) HHGYWEL 1 Phage display (common panning) 3-4 PMID:19923010 Anti-chicken IL-2 monoclonal antibody 6H10 Interleukin 2 Not determined. Not determined. Ph.D.-7 phage display library (X7) 653 HGEVPRFHAVHL(14) APWHLSSQYSRT(3) DLNYFTLSSKRE(3) SPLITSTLIPQR(2) TALATSSTYDPH(2) NRQSNWPIHKTI(1) QLSEECSYLISRP(1) QLTKNVPTYKSS(1) SNPQPYTILPPV(1) SPNQPYTILPPV(1) TPLTLRTQTLTQ(1) TTKQPHFHQKTL(1) TTLVSTGQRTHP(1) 13 Phage display (common panning) 3 PMID:20067240 Human embryonic stem cell line H9 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The library converged at the third round of screening, and 4-5 peptides dominated the sequence pool. A comparison of the number of unique sequences at each screening step revealed that convergence (such as HGEVPRFHAVHL) occurred not at the panning step but rather during amplification. Binding test was not done. 654 SNHPATLTGTG(7) TSPAYQATSPAP(5) APWHLSSQYSRT(5) SSLPPQQPSVSR(3) SLPKFSDLLPRP(2) ATQEIHASSALL(2) LSTHTTESRSMV(2) MPHLRSPSQDER(1) QSDSDDLTVFAL(1) QSQHTDPQYLYL(1) SISKTSHSHTQY(1) VQQKSSMVGPLL(1) 12 Phage display (common panning) 3 PMID:20067240 Human embryonic stem cell line H9 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The library converged at the third round of screening, and 4-5 peptides dominated the sequence pool. A comparison of the number of unique sequences at each screening step revealed that convergence (such as SNHPATLTGTG, APWHLSSQYSRT and SSLPPQQPSVSR) occurred not at the panning step but rather during amplification. Binding test was not done. 655 LTTAPKLPKVTR(1) QLGTQPNSRTYA(1) SRYITTMTNEQV(1) TVKHRPDALHPQ(1) LSTIGMPKLLA(1) VTSRTIIPQGSA(1) FAKSPDVSLNPS(1) 7 Phage display (common panning) 2 PMID:20067240 Human embryo carcinoma cell line NCCIT Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) phage display (biopanning and rapid analysis for selective interactive ligands, BRASIL panning, amplification-free panning) From 370 clones tested, seven cell-binding phage clones were identified and sequenced. Of the seven phage-derived peptides identified, all supported cell binding except FAKSPDVSLNPS. The self-assembled monolayers presenting LTTAPKLPKVTR were especially effective, as they supported cell adhesion over the broadest range of densities. 656 ETTKLQRFQAML(50) DHTLFAASHNHR(6) DHTLFSTGHSHG(2) DHTFMQRYHTHQ(1) 4 Phage display (common panning) 4 PMID:20177903 Anti-TSOL18 monoclonal antibody 17E1 Activated oncosphere TSOL18 Not determined. Not determined. Ph.D.-12 phage display library (X12) 657 DTPYDLTG(15) DTDSHVNL(9) ERAPLSVE(6) GDNSHVNL(5) VGQDSDYS(4) VSDNTDYS(3) GPDSTWAG(2) EGMMYTDV(2) DLTYVNSQ(2) DSSNKPTG(1) DSSRLERV(1) AHALTTEE(1) APLPTNGE(1) ATDHAAPQ(1) ATPTTPDP(1) EKFASNST(1) EVSMYTDV(1) 17 Phage display (subtractive panning) 4 PMID:20185523 Prostate carcinoma cell line PC3 Not determined. Not determined. Not determined. f8-8mer phage display library (X8) The naive library was sequentially exposed to culture flasks, serum treated culture flasks and non-target cells (fibroblasts) before being incubated with target PC3 cells. The sequenced phage clones were from eluate, post-elution wash and lysate. DTDSHVNL and DTPYDLTG demonstrated specificity and selectivity to PC3 cells based on target-association assays, microscopy and flow cytometry. 658 DVVYALSDD(14) AEYGESVNA(12) AEYGERGNA(6) AEYGESGNA(5) EGLVWIGMD(3) EAAGANIAP(2) AEYGESVLI(1) GAYDVNVND(1) DSDVGWVND(1) GPNWAEGDS(1) VADDRDYSD(1) VDVSEQMSL(1) VGDNVDYMD(1) VGDYDVVDS(1) 14 Phage display (subtractive panning) 4 PMID:20185523 Prostate carcinoma cell line PC3 Not determined. Not determined. Not determined. f8-9mer phage display library (X9) The naive library was sequentially exposed to culture flasks, serum treated culture flasks and non-target cells (fibroblasts) before being incubated with target PC3 cells. The sequenced phage clones were from eluate, post-elution wash and lysate. DVVYALSDD demonstrated specificity and selectivity to PC3 cells based on target-association assays, microscopy and flow cytometry. 659 VQASNSN(39) IPTHIRP(6) LPPPPNP(3) SPYRSLS(1) SQTPKSK(1) NQHTRVS(1) LPLTPLP(1) AVNSLQW(1) 8 Phage display (in vivo) 4 PMID:19481546 Heart of the normotensive Wistar Kyoto rat (WKY) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the WKY rat, the heart-homing capabilities of phage displaying VQASNSN, SPYRSLS and IPTHIRP were much higher than those of insertless phage. For example, IPTHIRP phage was 49.8-fold higher. 660 VQASNSN(60) LPLTPLP(31) DDTRHWG(9) LPPPPNP(4) IPTHIRP(3) AVNSLQW(1) 6 Phage display (in vivo) 3 PMID:19481546 Heart of the stroke prone spontaneously hypertensive rat (SHRSP) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the SHRSP rat, the heart-homing capabilities of phage displaying DDTRHWG and VQASNSN peptides were compared to insertless phage. VQASNSN homed to the heart at levels 200-fold higher than insertless phage. In addition, VQASNSN also homed to the lung, liver and kidney at levels more than 10-fold higher than insertless phage. DDTRHWG showed 46-fold enhancement in SHRSP rat compared to 11-fold in WKY rat further supporting its potential selectivity for the SHRSP heart. 661 YDPWTPS 1 Phage display (common panning) 3 PMID:19624458 von Willebrand factor, vWF Type I collagen Not determined. Not determined. Ph.D.-7 phage display library (X7) After three rounds of biopanning on a VWF-coated surface, 94 individual phage clones were grown, of which 10 bound to VWF. Eight clones displayed the same YDPWTPS peptide. Phages diplaying YDPWTPS bound to VWF in a specific and a dose dependent manner and specifically inhibited the VWF-collagen interaction under both static and flow conditions. YDPWTPS is the first peptide antagonist that binds to the VWF C domain and inhibits the VWF binding to collagen, suppressing platelet adhesion and aggregation under high shear conditions. 662 CFQHPSFIC(7) CSGYPRHYC(3) CRQSHLKVC(2) CKYNTHHAC(2) CTGSPELHC(1) CILGRIFSC(1) CNQHTALSC(1) CQMPEMKAC(1) CWPNLTKEC(1) CTVSTSIRC(1) CPASKTLTC(1) CLPSSGAAC(1) 12 Phage display (subtractive panning) 4 PMID:20186460 Lipopolysaccharide (LPS)-activated alveolar macrophages (AMs) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A strategy of sequential screening was applied by firstly binding the phage library to unmanipulated AMs and then binding the flow-through phages to LPS-activated AMs. After 4 rounds of panning, 22 out of 40 phage clones selected randomly bound selectively to LPS-activated AMs. The FQHPSFI peptide significantly inhibited LPS-stimulated microphage inflammatory protein 2 production in vitro. 663 YSLRADSRWMPS(8) KMDRHDPSPALL(4) FQPHMARKLPSL(4) TMAKSHMILEMA(1) GDASIRNQMWLF(1) TSNQNMTRLPPA(1) TMGFTAPRFPHY(1) S-x(2)-T-M-x-R-x(2)-P-S-L 7 Phage display (competitive panning) 5 PMID:20230853 Interleukin 2 IL-2 alpha receptor Not determined. Not determined. Ph.D.-12 phage display library (X12) The phages were eluted with chicken CD25. 664 TPDCTRWWCPLT(9) TTCNNWVCFFEW(3) MCTGKICWLNEM(3) HTACSKTLCPLP(3) VPHCSDTVCSLP(2) P-x-C-T-x-W-x-C-P-L-P 5 Phage display (competitive panning) 5 PMID:20230853 IL-2 alpha receptor Interleukin 2 Not determined. Not determined. Ph.D.-12 phage display library (X12) The phages were eluted with chicken IL-2. 665 LLADTTHHRPWT(0.5) SVSVGMKPSPRP(0.1) LLADTTHHRPWP(0.083) LLADATHHSPWP(0.083) HSVSNIRPMFPS(0.042) SVSEGTHPSPRP(0.042) 6 Phage display (in vivo) 6 PMID:20232090 Bevacizumab-treated tumor vasculature in nude mice Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Cancer therapy response to was bevacizumab evaluated in LS174T human colorectal cancer model, which was made on the right shoulder of female athymic nude mice. LLADTTHHRPWT was found to be dominantly enriched in the bevacizumab-treated tumors. 666 TNPNRRNRTPQMLKR(5) 1 Phage display (subtractive panning) 5 PMID:18472186 Beta-amyloid protein 42 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Subtractive steps were used to remove phages binding to streptavidin or rat Aβ. Thus, it can be used to select peptides that target the active site of human Aβ’s SOD like activity, a site not present in the highly homologous rodent sequence. Twenty individual phage clones were randomly picked and sequenced. However, only one sequence was given in the original paper. TNPNRRNRTPQMLKR significantly reduced Aβ42\'s SOD-like activity. This 15-mer peptide reduced Aβ42 neurotoxicity in a dose-dependent manner. Furthermore, comparative analysis of the 15-mer peptide with Clioquinol, a known inhibitor of Aβ’s metal-mediated redox activity, showed the 15-mer peptide to be equipotent to this metal chelator, under the same experimental conditions. 667 PLPQML(8) MTMPTM(5) 2 Phage display (subtractive panning) 5 PMID:18472186 Beta-amyloid protein 42 Not determined. Not determined. Not determined. f3-6mer phage display library (X6) Subtractive steps were used to remove phages binding to streptavidin or rat Aβ. Thus, it can be used to select peptides that target the active site of human Aβ’s SOD like activity, a site not present in the highly homologous rodent sequence. Twenty individual phage clones were randomly picked and sequenced. However, only two sequences were given in the original paper. PLPQML significantly reduced Aβ42\'s SOD-like activity. 668 STPNPLPRWLVP(4) ESSWQDMLYLLR(3) WKTQTHPFWDDL(2) QHTPHPLPPFLI(1) SHKQDPLPWPRF(1) KVQLPLPYPFPF(1) KHDPLPYPHFLL(1) HLWEHEMILDLL(1) THRWQEELTLLL(1) SGIHAYPHWLDH(1) 10 Phage display (subtractive panning) 3 PMID:19864424 Mitochondrial fission 1 protein Dynamin-1-like protein Not determined. Not determined. Ph.D.-12 phage display library (X12) At the end of the third round of panning, plate binders were eliminated by binding to a BSA coated plate. 669 FYSHSFHENWPS(26) IKSNPQTSNEFL(1) SAHGTSTGVPWP(1) ELANSVPWASPP(1) WSPGQQRLHNST(1) 5 Phage display (common panning) 4-5 PMID:19891006 Troponin I, cardiac muscle Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Thirty clones (20 from the 5th round and 10 from the 4th round) were sequenced. Kinetic indirect phage ELISAs revealed that FYSHSFHENWPS was found to have nanomolar affinities for for human troponin I while attached to the phage particles. However, the binding affinity was decreased by the presence of complex tissue culture media (MEM), and the addition of 10% calf serum further interfered with the binding of the target proteins. Coincidently, FYSHSFHENWPS was reported to be a BSA binder by SAROTUP. 670 LANKHQRLHAAD(2) FHSSWPVNGSTI(2) STYFDPHGTHPQ(2) HGDISQINSSPW(1) DRAPLIPFASQH(1) ASITHFKSGKSH(1) TMGFTAPRFPHY(1) NHVHRMHATPAY(1) QHANHQAWNNLR(1) WNRSPAFSGPYL(1) HHEWTHHWPPPP(1) TIPYHVPYKPPR(1) HKTQTYPQAWMH(1) 13 Phage display (common panning) 4-5 PMID:19891006 Troponin I, cardiac muscle Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Of 54 clones sequenced at 4th or 5th round, 16 readable sequences were obtained. Kinetic indirect phage ELISAs revealed that FHSSWPVNGSTI was found to have nanomolar affinities for for human troponin I while attached to the phage particles. However, the binding affinity was decreased by the presence of complex tissue culture media (MEM), and the addition of 10% calf serum further interfered with the binding of the target proteins. Coincidently, FHSSWPVNGSTI was reported to be a plastic binder by SAROTUP. 671 CKVALTLRC(5) CLVYLTQRC(3) CIVYLTQKC(3) CKLAMTMKC(3) CKVILTHRC(2) CLWFPREQC(2) CILYLTQKC(1) CFLVMSQRC(1) CMLPLYFPC(1) CELPRSPSC(1) CTVPAFPAC(1) CTNSAMADC(1) CKHEPTPNC(1) V-x-L-T-x-R 13 Phage display (subtractive panning) 3 PMID:20156289 Tumor necrosis factor receptor superfamily member 10B Tumor necrosis factor ligand superfamily member 10 1D0G,1D4V,1DU3, Not determined. Ph.D.-C7C phage display library (CX7C) In fact, the target used is DR5-Fc, fusion of the ecto-domain of the receptor to the Fc-portion of human IgG1. Before the third round, a subtractive round was performed by incubating the amplified phage of round two with human IgG Fc fragment. After three rounds of biopanning against DR5, 25 clones were picked and sequenced. Only 14 clones showed binding to DR5 with ELISA. These DR5-binding clones correspond to the following sequences: CKVILTHRC, CKVALTLRC, CLVYLTQRC, CIVYLTQKC and CILYLTQKC. The synthesized peptide, YCKVILTHRCY, in both monomeric and dimeric forms, binds specifically to DR5 in such a way that TRAIL binding to DR5 is inhibited. 672 CSPGGVVSVCIC(1) CFSGCGWVVKWC(1) CGERFPLVDPCC(1) CSLRGLDLRCFC(1) CWLPVVVGSRCC(1) CVLGGLSVVFEC(1) 6 Phage display (subtractive panning) 3 PMID:20214576 Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Not determined. CX10C P99 β-lactamase-fusion library The target used is the extracellular domain of ErbB2 receptor. Before panning, subtractive steps were taken with plastic and casein to remove non-specific binders. 673 MCGVCLSAQRWT(1) VSCPWLKYSGAL(1) VWGLPSCTLAHG(1) QSIFVDTMFRGS(1) DDLSYLMVPGLL(1) SGLWWLGVDILG(1) 6 Phage display (subtractive panning) 3 PMID:20214576 Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Not determined. X12 P99 β-lactamase-fusion library The target used is the extracellular domain of ErbB2 receptor. Before panning, subtractive steps were taken with plastic and casein to remove non-specific binders. 674 CGDGVWVGWVRC(2) CGDRLCRMLWLC(1) CGRVGMDVMGGC(1) CGLGWCGVRWGC(1) GGLHKDVCVAIF(1) CMGWLPWWRTHC(1) CPWLIRGLVGCC(1) CGIRWLAFPYGC(1) 8 Phage display (subtractive panning) 3 PMID:20219829 Breast cancer cell line BT-474 Not determined. Not determined. Not determined. CX10C P99 β-lactamase-fusion library Before panning, subtractive steps were taken with plastic and human non-cancer epithelial MCF 10A cells to remove non-specific binders. 675 LSVCVRGLLGCG(2) VCMNFVPAICRV(1) KFLAYPSFFSRC(1) LFMAGGSCYLSS(1) DRCWSILATSTF(1) HLLWVCPGGAPC(1) VWGVFGGCSQRP(1) LSVWMQGLSRSL(1) VLGMARWVDLGS(1) SVVSSVWRVSDS(1) 10 Phage display (subtractive panning) 3 PMID:20219829 Breast cancer cell line BT-474 Not determined. Not determined. Not determined. X12 P99 β-lactamase-fusion library Before panning, subtractive steps were taken with plastic and human non-cancer epithelial MCF 10A cells to remove non-specific binders. 676 TCSKKYPRSPCM(16) 1 Phage display (in vivo) 3 PMID:20223498 Pea aphid guts Not determined. Not determined. Not determined. LX-8 phage display library (XCX8CX) Briefly, aphids were allowed to feed for several hours through parafilm membranes on a sucrose solution containing the f88.4-LX8 phage library. The guts were then dissected, unbound phage washed away, and bound phage removed, amplified and used for another round of feeding. After each round of biopanning, between 100 and 400 phage plaques were recovered from the aphid gut epithelium. After the third round of biopanning, the phage DNA from 16 plaques was extracted, and the DNA sequences encoding the peptides displayed by each phage were determined. TCSKKYPRSPCM bound to the midgut and hindgut of the pea aphid Acyrthosiphon pisum. Binding was confirmed by labeling the aphid gut with a TCSKKYPRSPCM-green fluorescent protein fusion. TCSKKYPRSPCM reduced uptake of Pea enation mosaic virus (Luteoviridae) from the pea aphid gut into the hemocoel. TCSKKYPRSPCM also bound to the gut epithelia of the green peach aphid and the soybean aphid. 677 LALPPLAPNHHH(7) 1 Phage display (common panning) 3 PMID:20300752 Purified serum IgG from ankylosing spondylitis patients Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Seventeen out of twenty randomly selected phage clones exhibited specific reaction with purified sera IgG from ankylosing spondylitis patients, among them seven coming from the same clone. And only this clone's inserted peptide sequence was given in the original paper. 678 GHLHLRVPTLKM EFIRSPHSVDWL SQSRNPPMPPPR RRPPYRVPPKLF SLYERHPASTYP HTVSRRPLPSSG RHTHGNLLRFPP RKPTQSLPTRLV TRRPHKMRSDPL SRQFLHSLDRLP QTPYQARLPAVA RNNLNQTYPERR YSLLPVRPVALT HLHHHLDHRPHR TLTWHTKTPVRP WHPHRHHHLQWD R-x-P 16 Phage display (common panning) 3 PMID:16690883 Gap junction alpha-1 protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After removing the unbound viruses by washing with Tris buffered saline containing 0.5% tween-20 (TBS-T), low affinity binders were eluted with a solution of TBS-containing Cx43CT at a concentration of 100 μg/ml. High affinity binders were then recovered by overlaying the well with a culture of E. coli, to allow the tightly bound phage to infect the bacteria. The paper suggest that RXP-based peptides could serve as tools to help determine the role of Cx43 as a regulator of function in conditions such as ischemia-induced arrhythmias. 679 VSYSELTSYYMR(27) ISWMDLTAYYRG(2) 2 Phage display (subtractive panning) 3 PMID:16041387 Capsid protein p24, CA Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Bovine serum albumin(BSA) was used for negative selection to screen out peptides that target the nucleic acid–binding domain. After three positive and two negative selections, 36 of 41 analyzed phages bound to CA but not to BSA. 680 ITFEDLLDYYGP(15) ISWSELDAFMQM(6) VSYSELTSYYMR(6) STTWQDFFKTFG(5) YNEPWWLTPSMF(3) VKYHDLQTFFDP(1) VTYAQLQAYFPD(1) LEFSDLEDFFRA(1) LNFSDLNNYFLL(1) LDYPWWLSMNNI(1) SYTQWDNAPGTR(1) IADRPRAWIGSP(1) AMKTHTAIAPRA(1) HPQMHATPYQTT(1) SPSNLYEQLLHW(1) LPMIDIYRTAEL(1) 16 Phage display (subtractive panning) 3 PMID:16041387 C-CANC Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) NC was used for negative selection to increase specificity. After three positive and two negative selections, The peptide coding regions from 46 were sequenced and shown to code for 16 different peptides. 681 STFTKSP(0.50) NHWASPR(0.28) 2 Phage display (in vivo) 5 PMID:15536193 Murine bone marrow Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) BALB/c mice 6-8 weeks of age (Taconic Farms, Germantown, NY) were injected intravenously (tail vein) with PDPL (Ph.D-7 Phage Display Peptide Library Kit, New England Biolabs) diluted in 0.25\r\nml Dulbecco's modified Eagle's medium (DMEM). 682 STFTKSP(0.85) 1 Phage display (common panning) 3 PMID:15536193 Murine hematopoietic stem cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Biopanning combined with fluorescence-activated cell sorting (FACS) was repeated three times, yielding a phage clone that attached to primitive\r\nLin-Hoechst/rhodamine stem cells. 683 THRTSTLDYFVI SHKYPLPPYFHW TIKMHTLSYTGL 3 Phage display (common panning) 5 PMID:15895095 Chlorine-doped polypyrrole, PPyCl Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 684 SGHQLLLNKMPN(19) LWATFPPRPPWL(15) WSAAPTKPPYHT(5) ILANDLTAPGPR(4) HHGHSPTSPQVR(3) LPYGTSNRHAPV(3) YVQGWNYHDLTR(3) LWAAFPPQASVA(3) FDTPHTLTWFHG(2) 9 Phage display (subtractive panning) 6 PMID:15319030 Human glioma cell lines Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The library was screened against\r\na mixture of human glioma cell lines including dGli36, SF767, U87MG, U251MG, and U373MG at 37°C to increase the probability of obtaining sequences that could interact generically with most glioma cells. Phages rescued were further subjected to negative panning with nonglioma cell lines A549, CNE2, and\r\nHepG2, to eliminate nonspecific background binding. LWATFPPRPPWL is able to target specifically to tumors of glial origin, which would allow the design of applications related to the diagnosis and treatment of human gliomas. 685 ALPSTSSQMPQL ALPSTSSQMQL ALPSTSSQMPQV YQSSVSVQLPTL TTRQVPVSYTSS RLGFPPQTHAL HQPIQILEQPYT NSQNIGVGSW HVDQRYWFLGAP TTGPNTRHHA MGIAEQLMH 11 Phage display (subtractive panning) 8 PMID:16889372 Undifferentiated P19 embryonic carcinoma stem cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) To reduce nonspecific binding of phages to the cell surface, the phage libraries were preadsorbed to the differentiated P19 cells before each selection on the undifferentiated P19 cells. ALPSTSSQMPQL may be targeted to a marker expressed on the stem cell and thus become a practical tool for the isolation of somatic stem cells. 686 SNTQLDQ AAKTWNA HQLTKTL TELLVVT TGGSYGS LPGIPWL TLQPPLL SPNMNLT LTPHPIY NDTRPPR VPLNHSP QHSSSFY TMTWTSQ TTATDYS SAPKTTF 15 Phage display (common panning) 3 PMID:12882541 Quinoprotein glucose dehydrogenase Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 687 NATLTRL(3) VTYRAPA(2) HTLNWTH(1) HTPMLHP(1) QGIAEFR(1) QLHPLKP(1) LGAPFTG(1) 7 Phage display (common panning) 4 PMID:12882541 Quinoprotein glucose dehydrogenase Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 688 NATLTRL(7) SILPYPY(3) 2 Phage display (common panning) 5 PMID:12882541 Quinoprotein glucose dehydrogenase Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 689 NATLTRL(10) 1 Phage display (common panning) 6 PMID:12882541 Quinoprotein glucose dehydrogenase Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 690 LPAPLLPAAPLY(4) SISKSPMSLLSP(1) WAPGSMPTSRLA(1) GNYWSNGHSYHS(1) 4 Phage display (common panning) 4 PMID:12882541 Quinoprotein glucose dehydrogenase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 691 LPAPLLPAAPLY(9) 1 Phage display (common panning) 5 PMID:12882541 Quinoprotein glucose dehydrogenase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 692 GETRAPL(5) EYHHYNK(2) AAPMQVT(1) AKPSPFP(1) ALQBKPI(1) AMPYAPR(1) ANMSLLT(1) ANSKLSP(1) APATSIG(1) APQPWLM(1) ASTQQPT(1) DLRIAAS(1) EGLPANP(1) EYTHTPY(1) FPGKQTT(1) GHSHSHS(1) GPGPNIS(1) GPNQVEW(1) GSTQPPW(1) HLHTIGR(1) HPFILKP(1) HPPBVSS(1) HSFPHAP(1) HVLWTPP(1) IRPPSII(1) KLVASNP(1) KVTTTRV(1) LAKHPDS(1) LERGPYG(1) LVPPSGT(1) MGPPSTP(1) MPPGYPH(1) NALKFSA(1) NPFYSLR(1) QLTMFPS(1) QPNNHAH(1) QQQHPFK(1) SAPERFS(1) SFGENSI(1) SGSPPSV(1) SLPDPIH(1) SLRPSID(1) SMPKLIN(1) SNAQSMR(1) SNMAQHR(1) SPIRHVH(1) SPQLPQL(1) SQSPFFP(1) SSHGSLS(1) SSQYAHL(1) SWLPHNA(1) SYMYKPQ(1) TARQDSI(1) TGHHIFY(1) THLSRTP(1) TLSNYSQ(1) TNGLRTA(1) TPPQSTG(1) TPTIHKT(1) TPVQQVA(1) TQEYRSA(1) TQMRQPP(1) TQPPIRT(1) TSPIPPK(1) TTPRFIL(1) TTTLRPS(1) TYATDRR(1) TYSQSMT(1) VKPBTGA(1) VLPRASY(1) VNPVNTH(1) VSAQTRQ(1) WAPPPAG(1) WNLQPPQ(1) WPNTYRL(1) YHPMSSL(1) YLKPPGP(1) 77 Phage display (subtractive panning) 4 PMID:14759804 Human saphenous vein vascular smooth muscle cell Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Following one round of biopanning against target HSVSMC, a preclearing step was introduced whereby biopanning on nontarget cells (HepG2 cells and peripheral blood mononuclear cells) was performed. 693 ATPPPWLLRTAP DGSIHKRNIMPL DYDSLSWRSTLH GEPTTDMRWRNP GLWPWNPVTVLP HMLNDPTPPPYW KPAYTHEYRWLA LETTCASLCYPS LGTDWHSVSYTL LGTLNAGVPGFP LTHSKNPVFLST LVPTTHRHWPVT LVSNARGFNNLS NTRIPEPIRFYM NVYTFHSMSPMP QHTTLTSHPRQY SDFSDTMPHRPS SIDTIQILSLRS SISWASQPPYSL SMVKFPRPLDSR SPTLGASVAQTN TMSPNVYYTAFG TQIPSRPQTPSQ VCSNMYFSCRLS VPPHPMTYSCQY VPRLEATMVPDI VPTKPELPVNFT WSSDLPQPASTY YITPYAHLRGGN NVYTDNTLSPTP 30 Phage display (in vivo) 4 PMID:15139528 Gastrointestinal tract tissue, GIT tissue Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage display libraries were screened in vivo within the gastrointestinal tract of a rat model by successive screenings across four cycles of selection. LETTCASLCYPS and VPPHPMTYSCQY were shown to bind to receptors on the surface of human intestinal tissue. 694 FHNHGAA(3) FHNHGKQ(2) RLFTWE(2) SSLPLRK(2) FHNHGIL(1) FHNHGAT(1) FHNHGST(1) FHNHGAP(1) FHGHGLY(1) HIEHLIA(1) HLEHLLF(1) PFFHLIG(1) STPIPAP(1) STFTHPR(1) 14 Phage display (common panning) 3 PMID:15476403 Integrase, IN Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) FHNHGKQ and HLEHLLF exhibit a high affinity for IN and were chemically synthesized. IN. These IN-binding agents could be used as a base for developing new anti-integrase compounds as well as for structural studies of the still unknown three-dimensional structure of the entire integrase molecule. 695 NPRLYE(4) TTYSRFP(3) AEPVAML(2) VPTGYKP(2) ASSRTPS(1) HFWNRPL(1) FHQNWPS(1) HWGMWSY(1) HAWNYIF(1) NSHAIYP(1) SLLSSPQ(1) SPYHTQP(1) LPPNPTN(1) 13 Phage display (common panning) 3 PMID:15476403 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 696 SWALWMLG(14)[NT] FVRWLLLGVP(9)[NT] WLGLMRW(4)[NT] WARWLLLTPA(4)[0.329 ± 0.071] PWLGLWLLGV(1)[0.333 ± 0.058] PWLARWLLGV(1)[NT] LGCWILPRL(1)[NT] DTWARWML(1)[NT] W-L(0,1)-A-R-W-L(2)-[GL] 8 Phage display (common panning) 4 PMID:15672819 Harpin hrpZ Not determined. Not determined. Not determined. Three pools of fUSE5 phage display library ELISA Optical density was quantified at 492 nm. Data for both phages were combined of the results of four assays, each with at least four replicates of the protein sample. Data shown were reproduced from the graph. NT denotes not tested. 697 VPSSGPQDTRTT(4) YSPDPRPWSSRY(3) TLWSQGRSAYPV(2) NNRPEPSPVVPH(1) SPLDGKNIPLGH(1) WPAPAIWHAPTL(1) 6 Phage display (common panning) 3 PMID:15703906 Aluminum Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Most of peptides are rich in hydroxyl-containing amino acids, serine or threonine, or contain histidine. For the aluminum-binding peptides, peptides with a higher number of hydroxyl-containing amino acids bind to the aluminum surface more tightly. The paper suggests an important role for the hydroxyl-containing amino acids in the metal-peptide interaction. 698 ATIHDAFYSAPE(1) NLNPNTASAMHV(1) NLTIASYPSMVV(1) QSHYRHISPAQV(1) QMDISLGRWSSM(1) YMKQIPAGRTNP(1) 6 Phage display (common panning) 3 PMID:15703906 Mild steel Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Most of peptides are rich in hydroxyl-containing amino acids, serine or threonine, or contain histidine. The paper suggests an important role for the hydroxyl-containing amino acids in the metal-peptide interaction. 699 CTVALCNMYFGAKLD(25) GWCDVALCDPLLP(16) GWGDVALCDPLLP(3) PSYCPAPAVSIVAVC(2) V-A-[LV]-C 4 Phage display (common panning) 3 PMID:15789570 Anti-phomopsin A monoclonal antibody C3C11 Phomopsin A Not determined. Not determined. f3-15mer phage display library (X15) The paper suggests that the mimotope peptides bind to mAb C3C11 at the same site as phomopsin A. 700 CGPAEDRVRPWC(7) CNFDRTAPGQWC(5) CGAQMSRVRPAC(5) CSAAVSRTQPWC(4) CDFSRTRAGTWC(3) CGGPNSRVRPWC(2) CGGPDSRTRTWC(2) CEPAVSRTRPWC(1) CDFSRSRPGIWC(1) CEAPESRVVAWC(1) CHAGFGQAWHSC(1) CHPEDRRWLFC(1) CGFYRSGVGQWC(1) CGPDKDRVGPWC(1) CGQHITRVRPWC(1) 15 Phage display (common panning) 3 PMID:15846144 Anti-HMW-MAA monoclonal antibody 225.28S High molecular weight melanoma-mssociated antigen, HMW-MAA Not determined. Not determined. CL10 phage display library (CX10C) 701 LHYSYPVADP QGQSYPDXVV PWSYPPVXES AVAYPVVDXD SVSYPSE AVAYPVVED PFAYPVXPPP PPVSVAYPEP PWSYPVQESP LAPAYPLPPD LHYSYPVTGP GSRAYPGAAI IYSYPVTEQQ PLAYPLPQSE 14 Phage display (common panning) 3 PMID:15882053 Anti-AMA-1 monoclonal antibody 5G8 Apical membrane antigen 1 Not determined. Not determined. Min-23-R10 phage display library 702 RLHGSWWPYP LPYLAWLPSQ RLQELWDVHT RIPAAWFSSA RLEAAWLPLP 5 Phage display (common panning) 4 PMID:15882053 Anti-apical membrane protein 1 monoclone antibody 1F9 Apical membrane antigen 1 Not determined. Not determined. Min-23-R10 phage display library 703 VSPSWWRGPL RLPPLPLWPG 2 Phage display (common panning) 4 PMID:15882053 Apical membrane antigen 1 Not determined. Not determined. Not determined. Min-23-R10 phage display library 704 QFHVAFWWPA ASVWWLGPVR MYTLWPSYGR SFEFFPRFGF RFAKRWPPVR LQVWWLGPVR 6 Phage display (common panning) 4 PMID:15882053 Mitochondrial import receptor subunit TOM70 Not determined. Not determined. Not determined. Min-23-R10 phage display library 705 IKPVFGVQA IHPLFGFRPS VHPVFAIPWG ARPFFGMFEG YSGITMLPYN VGWPYVYSKG 6 Phage display (common panning) 4 PMID:15882053 Anti-Nef antibody Protein Nef Not determined. Not determined. Min-23-R10 phage display library 706 TQVLSFVPWK 1 Phage display (common panning) 4 PMID:15882053 Protein Nef Not determined. Not determined. Not determined. Min-23-R10 phage display library 707 LGPTCL SDHLCL 2 Phage display (common panning) 3 PMID:15896002 1,3-diketones 1-BSA and 2-BSA Not determined. Not determined. Not determined. YLK-(NNK)6 phage display library 708 SPFLGQ 1 Phage display (common panning) 3 PMID:15896002 1,3-diketones 1-BSA and 2-BSA Not determined. Not determined. Not determined. (NNK)6-YLK Library 709 LHLHRRQMRP QHRRRSRLLMQKLPN RSRRMTRRARARAA RRTTHSPSTKLRPYP PSRRKQ SRRSPYY QRRLTRTA TSSRRLIRLMRRTQY TPRNIMLRRPHPRRP IRRNPS PPKTRRRRQRNTKNT RRISTN RRQQ RRIIP RRHNTTTY 15 Phage display (common panning) 2 PMID:15944412 Phosphatidylserine, PS Not determined. Not determined. Not determined. f3-15mer phage display library (X15) 15 clones encoded peptides that contained contiguous arginine residues. Unlike other Ca2+-dependent PS-binding proteins, these peptides did not require Ca2+ for binding to PS, and the addition of Ca2+ did not alter the phospholipid specificity. 710 HSLKNSMLTVMA(10) SPHTTPMQMLAH(2) 2 Phage display (common panning) 4 PMID:16041565 Tom20 missing residues 2-28 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) One had partial identity (SMLTVMA) with a bacterial signal peptide from Toho-1, a periplasmic protein. The other had partial identity with a mitochondrial inner membrane glutamate carrier. The bacterial signal peptide could carry a protein into mitochondria both in vivo and in vitro. The first six residues of the sequence, SMLTVM, were necessary for import but the two adjacent arginine residues in the 30-amino-acid leader were not critical for import. The signal peptides of Escherichia coli β-lactamase and Bacillsus subtilis lipase could not carry proteins into mitochondria. Presumably, the Toho-1 leader can adopt a structure compatible for recognition by the import apparatus. 711 HYTYWWL(10) 1 Phage display (competitive panning) 3 PMID:16196489 [PA63]7 complex Protective antigen, PA Not determined. Not determined. Ph.D.-7 phage display library (X7) Non-interacting phage were washed away, and then PA was added to elute phage that specifically bound PA. The series of truncated peptide mutants was used to identify a minimal peptide sequence, TYWWLD, that can be used to develop potent polyvalent inhibitors of anthrax toxin. 712 SSPWMRE(7)[2.325, 3.5] ELWRPTR(6)[2.659, 10] ARPHLSF(6)[2.474, 6.2] TLHLSPA(3)[2.763, 5.4] QLQKYPS(2)[2.615, 8.5] QTMTYSR(2)[2.546, 5.0] NLQEFLF(2)[1.988, 0.69] AAQTSTP(1)[1.912, 4.9] GIRHTNR(1)[2.156, 2.4] RPTR 9 Phage display (common panning) 5 PMID:16201785 Isotactic poly(methyl methacrylate), st-PMMA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The absorbance data were converted to affinity relative to that of a phage library for st-PMMA. Besides, the apparent affinity constants (Kapp, e11 1/M) were obtained. All data shown were reproduced from the graph. The paper report for the first time a peptide motif that specifically binds to a stereoregular polymer, isotactic (it) poly(methyl methacrylate) (PMMA). 713 HSKYPLPPLPSL(11) AHHHWHPLPTLP(5) HPGYPLPPFPLP(1) HTWHPLPILPPK(1) H-x-W-[YH]-P-L-P(2)-L-P 4 Phage display (common panning) PMID:16274251 SH3 domain of tyrosine-protein kinase Lck Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In fact, the target is glutathione s-transferase (GST)-SH3 fusion proteins. 714 CTKNSYLMC(3) CLSSSLSDC(3) CKNSLTMAC(2) CTTTDLRAC(2) CPVSLQALC(2) CSSTTPNAC(1) CNSTKYNQC(1) CTNTMLPQC(1) CDRHNLTFC(1) CPPSKMSQC(1) CQPALQMKC(1) CIPTHPRLC(1) 12 Phage display (subtractive panning) 4 PMID:16763842 HUVECs co-cultured with gastric cancer cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Immortal gastric epithelium mucosa cells GES and wild-type HUVECs were used as the absorber cells for whole-cell subtractive screening. 715 VYCVL FLFGLFFFTCRS FRIVTCATCVILKAFMGWHYHISRSRS CDSFARSCVRGVGIMKACGRTRVTS LFTDMALSGKVLVKACGRTRVTS LFLCCMGFHGPL LLLSHVSLVCRL CFLLSFVVLGYD 8 Phage display (common panning) 3 PMID:17979224 Paclitaxel (Taxol) Transcriptional repressor NF-X1 Not determined. Not determined. X12 T7 phage display library Among them, two phage clones included the same consensus amino acid sequence (KACGRTRVTS). Analysis of the protein database using BLAST revealed that a portion of this sequence is conserved in the zinc finger domain of human NFX1. 716 HDIYPRH(1) THRLLL(1) MPHIHRH(1) HPRPRNN(1) QGHIGNE(1) SPPQSRA(1) APPXSPT(1) YPXPPLX(1) RQGGQYP(1) QRATPFP(1) 10 Phage display (common panning) 3 PMID:17630779 Anti-N domain of human ACE monoclonal antibody 5F1 Angiotensin-converting enzyme, ACE Not determined. Not determined. Ph.D.-7 phage display library (X7) All sequences fall into two groups. Alignment of these sequences with each other and full-size ACE falls within amino acid residues 1256-1265 (in the cytoplasmic tail of ACE), whereas alignment with the N domain alone reveals similarity with amino acid residues 471-483. 717 QPHTLSMHRHYY(2) SPSDRLMHNHYH(2) TPPTSRAHAHYY(2) RNHQTHRRQDGK(1) FAWXQHLHALEP(1) 5 Phage display (common panning) 3 PMID:17630779 Anti-N domain of human ACE monoclonal antibody 5F1 Angiotensin-converting enzyme, ACE Not determined. Not determined. Ph.D.-12 phage display library (X12) All sequences fall into two groups. Alignment of these sequences with each other and full-size ACE falls within amino acid residues 1256-1265 (in the cytoplasmic tail of ACE), whereas alignment with the N domain alone reveals similarity with amino acid residues 471-483. 718 HITSLLS(6) HTTHREP(5) APASLYN(4) NQDVPLF(3) TTNGYI(3) AYPYPHT(2) NNATLG(2) NSLMRPA(2) QGFGSPL(2) THHPHQK(2) TLARPSV(2) WDPPLRL(2) WHRAPLP(2) 13 Phage display (in vivo) 2-3 PMID:17551506 Wistar Kyoto (WKY) rats' kidney Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Twelve- to thirteen-week-old WKY rats (n =3 rats/phage display round) were infused with 2e11 plaque-forming units (PFU) of PhD 7mer library in round 1, and with 2e11 PFU of amplified phage from the kidney for rounds 2 and 3. APASLYN were found in the kidneys and non-target organs(heart, brain, and lungs). Authors engineered the HI loop of Ad19p to accommodate peptide insertions and clones. Intravenous delivery of each peptide-modified virus resulted in selective renal targeting, with HTTHREP and HITSLLS-targeted viruses selectively transducing tubular epithelium and glomeruli, respectively. 719 CQSHKLPSC(2) CVGNDNSSC(1) CKPETAPLC(1) CKLPVPPHC(1) CEHPPYLQC(1) CTSSDHLQC(1) CIAPTHNLC(1) CKTHHNLIC(1) CPDALPSSC(1) CSTLAFPLC(1) CQSALTGVC(1) CTTLQSAQC(1) CPFSLSALC(1) 13 Phage display (in vivo) 3 PMID:17384922 Tumor endothelial cells, TEC Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Mice (n=4 per round) were injected intravenously with 1e11 PFU of the whole phage library. After 15 min of circulation time, the animals were killed, and the number of phages localizing to TEC formed vessels as well as to murine kidney (control murine tissue) was determined. No such enrichment was seen in the mouse kidneys, suggesting the isolation of specific phages that preferentially bound to human tumor vessels rather than to mouse normal vasculature. 720 CTVALPGGYVRVC(9) 1 Phage display (subtractive panning) 5 PMID:16878978 Human melanoma cell line Not determined. Not determined. Not determined. CXnC phage display library pool (n = 3-12) The highly metastatic melanoma clone Me6652/4 and the low metastatic clone Me6652/56 were chosen as positive and negative cell lines, respectively. A subtractive panning strategy was applied, in which the phage were first adsorbed onto reference cells before the supernatant of this reaction was transferred to the target melanoma cells, Me6652/4. DNA sequencing of the 15 clones revealed that 9 clones were identical. The corresponding predominant peptide sequence was the cyclic 13-mer CTVALPGGYVRVC, Pep42. The cellular receptor for Pep42 was identified as the surface membrane form of glucoseregulated protein 78 (GRP78), a member of the heat shock protein family and a marker on malignant cancer cells. 721 CHKKPSKSC(22) CTKRNNKRC(1) CRRWESKRC(1) 3 Phage display (common panning) 6 PMID:16841905 Silicon dioxide, SiO2 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Two of the phage surface displayed peptides enriched with basic amino acid residues, STB1 (HKKPSKS) and STB2 (TKRNNKR), showed a cross binding affinity to both metal oxides (SiO2, TiO2). 722 CHKKPSKSC(22) CTKRNNKRC(1) CDQQTSEFC(1) 3 Phage display (common panning) 6 PMID:16841905 Titanium dioxide, TiO2 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Two of the phage surface displayed peptides enriched with basic amino acid residues, STB1 (HKKPSKS) and STB2 (TKRNNKR), showed a cross binding affinity to both metal oxides (SiO2, TiO2). 723 RMWPSSTVNLSAGRR(4) RLAYWCFSGLFLLVC(2) PNLDFSPTCSFRFGC(1) GRVPSMFGGHFFFSR(1) PVAAVSFVPYLVKTY(1) 5 Phage display (common panning) 4 PMID:16536485 Complement receptor type 2, Cr2 Complement receptor type 2, Cr2 Not determined. Not determined. f3-15mer phage display library (X15) To select specifically bound phage and exclude nonspecifically bound phage, a stringent biopanning process was performed including several measures: extensive washing with TPBS (pH 7.4), stringent washing with TPBS (pH 5.0), decreasing the incubation time in each round of selection, and elution with sCD21 at the last round selection. Two sequences, PNLDFSPTCSFRFGC and RLAYWCFSGLFLLVC, contained two cysteine residues. The distance between the two cysteines are five and eight amino acid residues, respectively. Consequently, the formation of cyclic peptide structures with constrained conformations is more probable than cross-linking. Except for RMWPSSTVNLSAGRR, all sequences of selected peptides, consisted mostly of hydrophobic residues, which may participate in hydrophobic interactions with the CD21 receptor. However, the five sequences did not show obvious homology with each other. 724 SMSIASPYIALE(18) SMSIGSPYITFG(6) SPGPMKLLKTPL(6) SMSIASPYIPWS(5) TLNINRLILPRT(5) 5 Phage display (subtractive panning) 3 PMID:17043801 Human gastric cancer cells GC9811-P Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage selection was performed with the GC9811 cell line used for the depletion. GC9811-P cells were derived from its parental cell line, GC9811. No complete homologies were found with previously determined protein sequences using a GenBank search of short peptide sequences or nucleotide sequences. 725 IVNWNLTAPGPRFES(15) SEVRASERTAHYEH(13) VHHVGYRVPGPRFES(5) MWNMDPIWIAAVES(4) FGSSKGRAPGPRFES(4) WSNPFPGPAGES(3) DPEVAARELGPPVES(2) WWRLDPINVAAGES(2) SQYEDPFNVARRES(1) QLWWDPIGVARVES(1) YEFDPILISRVES(1) VPEVALKECGPPVES(1) QIEVALKELGPPVES(1) AIVTALIELGPPVES(1) AIQRAFQELGPPVES(1) AIRQAWLELGPPVES(1) AIMRSAQELGPPVES(1) WYIGKETHLGPPVES(1) WYIGPENNLGPPVES(1) GVRIDERELGPPVES(1) AVIRAQWELGPPVES(1) EPGRMVEALGPRFES(1) SEVAGFVVSDRW(1) TMRVDERELGPPVES(1) QDRLTRYVGPPVES(1) SLIGSGSWWPAVES(1) LINEATGPAFVES(1) QDITASFILGPPVES(1) 28 Phage display (common panning) 2-3 PMID:15777347 Anti-gp91-phox Monoclonal antibody CL5 Cytochrome b-245 heavy chain Not determined. Not determined. J404 phage display library (X9) Phage display analysis confirmed this specificity, indicating that the CL5 epitope contains the region 135-DPYSVALSELGDR of gp91-phox. 726 CRNCTVIQFSC CFQPLTPLCRC CQSYHELLLQC CQIPQRTATRC CSVRQGPVQKC CSSCQNSPALC CAKQRTDPGYC CWMSPRHLGTC CHYIAGTVQGC CPLVSLRDHSC CKQSYLHHLLC 11 Phage display (common panning) 3 PMID:15466209 HPV-16 E7 oncoprotein (28-LNDSSEEEDEI-38) Not determined. Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) 727 FTGCSRLLYGWCSLN AWTCLFSTGGICQSS TLICMVKIQVKCPMQ PTWCLEFMITACIRG SAVCVSFENFMCIGP 5 Phage display (common panning) 5 PMID:15996026 PreS region of the large surface antigen (LHB) Not determined. Not determined. Not determined. X3CX7CX3 phage display library 728 QEKIRVRLSA 1 Phage display (common panning) 3 PMID:15980334 TG1 Escherichia coli cells, E.coli Not determined. Not determined. Not determined. X10 phage display library The sequence QEKIRVRLSA showed features common to a variety of antimicrobial peptides: a net positive charge (
2) and a total residue hydrophobic ratio of 40%. However, no obvious similarity to other sequences was found in the Antimicrobial Sequences Database. 729 CIVPWGRYC(6) CLIPWGRFC(4) CLVPWGRYC(3) CAVPWARYC(3) CAVPWGRLC(3) CLVPWARWC(2) CKVPWARWC(1) CLVPWGRLC(1) CNVPWGRYC(1) CDVPWRDLC(1) CVPWRDWTC(1) CIPWGRYFC(1) CAVPWGRYC(1) CIVPWARYC(1) 14 Phage display (common panning) 5 PMID:15946218 Integrin alpha-IIb-beta-3, integrin αIIbβ3 von Willebrand factor (vWF) Not determined. Not determined. CX7C phage display library A CX7C phage display library was used to search for peptides that bind to integrin αIIbβ3 in the presence of GRGDS. Sequencing of the bound phage revealed only four RGD-containing sequences (CWARGDFRC, CEPRGDWRC, CVARGDWRC, CWARGDPRC). Consistent with \r\nselection strategy the majority of sequences lacked RGD and revealed novel binders. The consensus in these sequences was the tri-amino acid motif Val-Pro-Trp (VPW). 730 VGSLR[481000] RQTND[415000] NQRSS[388000] LQRAI[367000] QRLRD[307000] PDRHM[243000] LSGGR[207000] TVDYA[134000] LSRDN[127000] RGKTN[80000] NNKLR[74000] MQVKH[34000] TTDLR[27000] RVTST[26000] AYGYK[24000] STKGI[20000] KLKET[19000] VGLYD[18000] VVMKD[15000] RVDTG[15000] GHRIN[12000] YQSLN[12000] SDKVY[9000] HETLK[9000] TSYLN[9000] MQATK[8000] EAPAK[8000] PVHLY[7000] QPNGY[6000] AYGLA[6000] YQNSS[6000] SAVRP[5000] 32 Phage display (common panning) 6 PMID:15843175 Kallikrein-14, hK14 Not determined. Not determined. Not determined. Substrate phage display library based on a modified pH0508b phagemid Fluorescence The CFP fluorescent substrate assay has been utilized to determine the kinetics of peptide substrate selection from a phage display library. The kinetic parameter kcat/Km (1/M × 1/s) was determined and shown. In fact, the target is recombinant hK14. A SwissProt database search with selected sequences identified six potential human protein substrates for hK14. Two of them, laminin α-5 and collagen IV, which are major components of the extracellular matrix, have been demonstrated to be hydrolyzed efficiently by hK14. 731 SSRFESLFAGEKESR SSKFAALWDPPKLSR SRWQALFDDGTDTSR SRWAEVWDDNSKVSR SSNTPRFKEYFMQSR SRFADFFRNEGLSGSR SSRGLLWDLLTKDSR 7 Phage display (common panning) PMID:15328534 Ligand-binding domain of androgen receptor(AR) Not determined. Not determined. Not determined. X15 phage display library Binding affinities were measured for a selected set of these peptides and their interactions with AR determined by X-ray crystallography. Structures of AR in complex with FxxLF, LxxLL, FxxLW, WxxLF, WxxVW, FxxFF, and FxxYF motifs reveal a changing surface of the AR coactivator binding interface that permits accommodation of both AR-specific aromatic-rich motifs and canonical leucine-rich motifs. 732 PLSQLTFSHLGP PLSQLTDSHLGP NLSRHTYSHPHM LSLNQSSALTRL QTSLYENIGLLR AQPNWTSLRSLP HRSPHFSRHGLL HLAHHFHNRLKL PWTQHKAMSQM YTKASIFTVQPM 10 Phage display (common panning) 3 PMID:15337767 Interferon-binding domain of human p300 Cellular tumor antigen p53 Not determined. Not determined. Ph.D.-12 phage display library (X12) Thirty clones were tested, revealing consensus sequences with over 50% homology to the conserved BOX-I motif within p53. 733 SCPGEDCHTMYPFTYKGY(14)[0.229] TCPGEDCHTMYPFTYRGY(11)[0.197] RCQFSVWSWESLRCEFPY(2)[0.861] VCHAGELPFTFRGVPFPY(2)[0.185] TCPGEDCHSMYPFTYRGY(1)[0.259] RCHQNGVCMHRNVLLTTY(1)[0.149] RCDLYPDGNLWGMPCPTY(1)[0.185] NCNPLEKWCPTVEQTPWY(1)[0.780] 8 Phage display (common panning) 3 PMID:10725425 Anti-VP2 monoclonal antibody 2F2 Outer capsid protein VP2 Not determined. Not determined. XCX15 phage display library ELISA Absorbance at 450 nm were measured. 734 YLTMPTP[0.13] WPTPPYA[0.09] TPHNTVS[0.07] SLPAHAR[0.10] HSSLQTP[0.06] YSIPKSS[0.10] ALQPRYL[0.15] 7 Phage display (subtractive panning) PMID:10747021 Kinase domain of VEGFR2 Vascular endothelial growth factor A, VEGF-A Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Absorbance at 490 nm was measured. Data shown were reproduced from the graph. Chinese hamster ovary (CHO) cells expressing a recombinant KDR at their membrane surface were tested for their ability to bind VEGF in a variety of conditions. The last amplified eluate was absorbed twice on non-recombinant CHO cells to enrich for phage clones specifically binding KDR. At the end of the selection, 24 clones were isolated and analyzed. DNA sequencing showed that seven independent peptides had been selected, with no sequence homology. 735 ATWLPPR[1.01] NPRALNY[0.54] ANLFKAK[0.54] YHSSFQA[0.57] ILDNYKL[0.65] LPPNPTK[0.96] YAIMPLV[0.71] 7 Phage display (common panning) 3 PMID:10747021 Anti-human VEGF monoclonal antibody Vascular endothelial growth factor A, VEGF-A Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Absorbance at 490 nm was measured. Data shown were reproduced from the graph. 736 SSNHQSSRLIELLSR GSEPKSRLLELLSAPVTDV VESGSSRLMQLLMANDLLT HPTHSSRLWELLMEATPTM QEAHGPLLWNLLSRSDTDW HVYQHPLLLSLLSSEHESG HVEMHPLLMGLLMESQWGA GHEPLTLLERLLMDDKQAV LPYEGSLLLKLLRAPVEEV SGWENSILYSLLSDRVSLD AHGESSLLAWLLSGEYSSA GVFCDSILCQLLAHDNARL HHNGHSILYGLLAGSDAPS LGERASLLDMLLRQENPAW SGWNESILYRLLQADAFDV PSGGSSVLEYLLTHSTSIL PVGEPGLLWRLLSAPVWRE S-R-L-x(2)-L(2), P-[AVLIFYW]-L-x(2)-L, [ST]-[AVLIFYW]-L-x(2)-L(2) 17 Phage display (common panning) 3 PMID:10567548 Estrogen receptor, ER Not determined. Not determined. Not determined. (X)7LXXLL(X)7 library Peptides were classified into three different classes based on sequences flanking the conserved LXXLL motif. Peptide #293 was obtained in a similar manner from random peptide libraries. 737 FHYEIWIPPHRG(6) SSPWWLVSFTST(3) SWMTTPWGFLHP(2) SHSLIWRIPLLH(2) IYVPWYYAENLP(2) YNYSWNGVVFVP(1) AQSTPLMKPQKS(1) DQTTLQRFLGSH(1) QTIKPPITVHPS(1) QYNHILGYLPFQ(1) IMDPQNSKVTVA(1) LPIQNAKRSMVS(1) IMSPWDESFWNY(1) ASESYVLFPGTR(1) SNWHGPLSYQLM(1) ALPLQDTAATLS(1) QEIYLTPRGPQQ(1) IDRTQMWRQSDL(1) INRDHPLHAGQP(1) HQTPQSLARWSL(1) HSLRAIQLITGM(1) LPSHHHHRVPAA(1) IPTYHHHHPSLR(1) QMTHHHTHRPPI(1) DLHSHHHGHMNH(1) SMHHHHRPASPT(1) WIGDAKSSLHHA(1) HNHPHTTSHVSM(1) HNSIIYHWHTLP(1) HFNHNHRGFHLI(1) AASPHYSSSHSH(1) 31 Phage display (common panning) 4 PMID:10619429 60 kDa chaperonin Not determined. Not determined. 1DKD, Ph.D.-12 phage display library (X12) The authors have determined the crystal structures of the complexes formed by the most strongly bound peptide with the isolated apical domain, and with GroEL (PDB ID: 1DKD). The peptide interacts with the groove between paired α helices in a manner similar to that of the GroES mobile loop. 738 LNTKWLRGD(10) LPNPWVRGD(9) RDCAWCRGD(6) LKNPWQRGD(3) VNNKWARGD(3) VNTKWLRGD(3) KECAWCRGF(3) DWKARREG(3) YPGWPRKDL(3) YKNPWIRGM(2) WNKAWVRGS(2) FRCSWCRGE(2) GECRWCKGD(2) RIVRGVGGT(1) LNKAWVRGS(1) NANPWSRGF(1) LTNKWMRGD(1) IGGPWRRGF(1) INKAWVRGM(1) VSSPWVRGG(1) YISPWVRGF(1) AQSSWVRGN(1) VGGSWVRGS(1) GVRWWG(1) TSHWVRDTS(1) GWIKYRLEG(1) YPGWGRNDA(1) YPGWDRADK(1) YPGWPRSDY(1) [GAVLIMP]-[NK](2)-[PK]-W(0,1)-VRG-[DE] 29 Phage display (common panning) 3 PMID:11027685 Anti-Cyt b monoclonal antibodies 7D5 Cytochrome b-245 Not determined. Not determined. J404 phage display library (X9) The antibody 7D5, secreted by the hybridoma, bound to solubilized cytochrome b of the neutrophils but not to other proteins such as emoglobin, myeloperoxidase, and pig cytochrome P-450. 739 GPLGLS[+++] GPLGLK[+++] VRPLGI[++] APLGMS[+++] PLSISG[+++] PLQFRG[+++] PLTMMG[+++] PLPMRM[++] APLAMR[+++] PLSIQD[+++] PLSFQG[+++] VLPLPM[+++] PLSLLS[+++] PAGLSD[+++] PRGLVA[+++] PKGLRA[+++] PYGMRA[+++] PVALKA[+++] PKGVYS[+++] PKGITS[+++] PSGIHV[+++] PFGFKS[+++] GPPMSL[+++] PINLHG[+++] PHPLFL[++] PSELKG[++] MAPYAL[+++] PFAFQG[+++] PSAYHS[+++] PMSYNA[+++] PAEIVA[+++] PMEMVE[++] VTPYNM[+++] PPRAIR[+++] PRPFNY[+++] PLGMRG 35 Phage display (common panning) 2-5 PMID:10906330 Collagenase 3 Not determined. Not determined. Not determined. fTC-LIB-N6 phage display library Western blot +++ strong cleavage; ++ some cleavage; + less cleavage; ND no cleavage of phage by human collagenase 3 detected by Western blotting. The selected phages were shown to be more sensitive by Western blot analysis to human collagenase 3 treatment than the positive control phage fTCol-3S (Western blotting result, [+]) with the sequences of PLGLWAR and the negative control phage fTCol-3R (NPVEPA) (Western blotting result, [ND]). Phage display screening also selected five substrate sequences that share sequence homology with a major MMP cleavage sequence in aggrecan and seven substrate sequences that share sequence homology with the primary collagenase cleavage site of human type II collagen. In addition, putative cleavage sites similar to the consensus sequence are found in human type IV collagen. 740 TLIPRSFCPTHDRDC(6)[0.37] TARVFLLGSCRSCGP(2)[0.20] VRLDHGVWGSRLSLP(2)[0.37] SSSSFVLWLLRPGFS(2)[0.65] LVWISPPPGLFASFV(2)[0.23] WTSPWLLDYPVPSGA(1)[0.09] ASLMLLGGTSGPVTH(1)[0.16] RSKGVKFNLSGVFQA(1)[0.17] L-W-L(2) 8 Phage display (common panning) 3 PMID:11058544 Human spermatozoa Zona pellucida sperm-binding protein 3 Not determined. Not determined. f3-15mer phage display library (X15) ELISA Clones M1–M12 and a random unselected clone MR2, used as a control, were added to the plates coated with paraformaldehyde-fixed sperm. After repeated washing bound phage were detected by incubating with an anti-M13 phage antibody conjugated to HRP and measurements carried out at 450 nm. The A450 value of the control phage clone MR2 was 0.05 and clones with A450 less than 0.05 represent non-binders. Data shown were reproduced from the graph. SSSSFVLWLLRPGFS displayed a high level of affinity for the sperm surface and showed sequence homology with a dominant human ZP3 epitope (hZP 25-33). 741 HWPRPDDSFWRP(10) TSNFINRMNPGL(4) TLWSTSPLTNKL(2) TLSRITVSTFTH(2) NKWPLAHSQKKR(2) L-W-L(2) 5 Phage display (common panning) 3 PMID:11058544 Human spermatozoa Zona pellucida sperm-binding protein 3 Not determined. Not determined. Ph.D.-12 phage display library (X12) 742 RINNIPWSEAMM(8)[0.0087 ± 0.0021, 0.0061 ± 0.0016, 0.0094 ± 0.0005][-8.90] CHPLTPWEC(6)[>500, >500, >500][383.90] TSPYEDWQTYLM(5)[>500, >500, >500][463.14] FWHPMHD(4)[>500, 68 ± 4, >500][94.28] SWPWDTW(3)[>130, 21, ND][74.36] DTHRWPWYISQE(1)[500, 259 ± 12, 348 ± 3][88.35] SPWYSDYTRYLW(1)[>140, >140, ND][373.94] SDPYKLWASYMY(1)[>100, >100, ND][528.60] QPASNMTLGRGL(1)[NT][NT] SSVDESTAQVNW(1)[NT][NT] 10 Phage display (competitive panning) 8 PMID:10364331 Envelope glycoprotein gp120 (SU) Anti-gp120 monoclonal antibody 17b 1G9M,1G9N,1GC1,1RZJ,1RZK,2NXY,2NXZ,2NY0,2NY1,2NY2,2NY3,2NY4,2NY5, Not determined. Ph.D.-12, Ph.D.-7 and Ph.D.-C7C phage display library pool ELISA,Surface plasmon resonance (SPR) The inhibitory capacities of the identified peptides were tested with gp120 from an additional T-tropic strain (HXB2 and SF2) and an M-tropic strain (ADA). IC50s (μm, means ± standard errors) for inhibition of the 4dCD4-gp120 interaction was measured by competition ELISA. NT, not tested. ND, not determined. Besides, surface plasmon resonance (SPR) binding assays were performed on a Biacore (Piscataway, N.J.) instrument. The MAb 17b was coupled to a CM5 sensor chip. MAb 17b was dissolved in 10 mM acetic acid-sodium acetate buffer (pH 4.9). The surface was regenerated twice with 5 ml of 4.5 M MgCl2 after gp120 binding. Sequential injections (20 μl) of HXB2-gp120 (200 nM) with peptide at increasing concentrations (0.2 to 200 μM) were made over a surface with 1,500 RU of coupled MAb 17b. The amount of gp120 bound (expressed as surface plasmon resonance units [RU]) was measured 10 s after the end of the injection and by subtracting RUs bound for the same solution on a blank mock-coupled surface. Results are given as the percentage of gp120 bound relative to gp120 bound in the absence of peptide: 100 + 100 × [(RU - RUgp120)/RUgp120]. The data shown in the second square bracket were reproduced from FIG. 4 in the reference. In the first round of selection, bound phage were eluted with glycine of BSA. In subsequent rounds of selection, phage were eluted with solution of gp160(502-516) in PBSTB. RINNIPWSEAMM inhibit the interaction between gp120 and both four-domain soluble CD4 (4dCD4) and monoclonal antibody (MAb) 17b, a neutralizing antibody that covers the chemokine receptor binding surface on gp120. 743 PCQRAIFQSICN[0.40] 1 Phage display (in vivo) 4 PMID:11687659 Mosquito salivary gland Plasmodium Not determined. Not determined. LX-8 phage display library (XCX8CX) ELISA The absorbance at 450 nm was measured. Data shown were reproduced from the graph. For selection of phages that bind to salivary glands, 10 adult Anopheles gambiae females were dissected 30 min after injecting about 1.0e11 phages into their hemocoels. After the fourth round of selection, over one-third of the phages recovered from salivary glands displayed PCQRAIFQSICN. This peptide strongly inhibited Plasmodium invasion of salivary gland and midgut epithelia. 744 PCQRAIFQSICN[0.24] 1 Phage display (in vivo) 4 PMID:11687659 Mosquito midgut lumen Plasmodium Not determined. Not determined. LX-8 phage display library (XCX8CX) ELISA The absorbance at 450 nm was measured. Data shown were reproduced from the graph. For selection of phages that bind to the midgut lumen, phages (1e15/ml) suspended in 1 mg/ml of gamma globulin/120mM NaCl/20mM NaHCO3 were fed to the mosquitoes, and midguts were dissected 30 min later. After the fourth round of selection, almost half of the phages from midguts displayed PCQRAIFQSICN. This peptide strongly inhibited Plasmodium invasion of salivary gland and midgut epithelia. 745 TDITLF(18) FHFYAF(2) 2 Phage display (common panning) 3 PMID:10967104 TrpE protein Not determined. Not determined. Not determined. f3-6mer phage display library (X6) The TrpE proteins immobilized using anti-TrpE antibodies and protein A-Sepharose beads. 746 AARNSP(14) IARIRV(6) [LI]-A-[RK]-I-R 2 Phage display (common panning) 3 PMID:10967104 p85-N-SH2 domain Serine/threonine-protein kinase A-Raf Not determined. Not determined. f3-6mer phage display library (X6) The TrpE-p85-N-SH2 fusion proteins immobilized using anti-TrpE antibodies and protein A-Sepharose beads. 747 AIARIR(11) LARIRS(7) AKIRF(2) FTHGLA(1) [LI]-A-[RK]-I-R 4 Phage display (common panning) 3 PMID:10967104 p85-C-SH2 domain Serine/threonine-protein kinase A-Raf Not determined. Not determined. f3-6mer phage display library (X6) The TrpE-p85-C-SH2 fusion proteins immobilized using anti-TrpE antibodies and protein A-Sepharose beads. 748 WANANY(10) LDSFYF(3) GDYTLF(2) TRDSFN(1) EPGLRF(1) SEDGHF(1) AVRGYR(1) GRIDYA(1) FHAFAL(1) TWASGA(1) 10 Phage display (common panning) 3 PMID:10967104 PLCγ1-N-SH2 domain Not determined. Not determined. Not determined. f3-6mer phage display library (X6) The TrpE-PLC The sequence GDYTLF is selected by each of the SH2 domains of GAP. 749 GDYTLF(9) RWYIDD(2) WLEVGK(1) SWAVNR(1) QASPIL(1) VRELVR(1) RYVFNH(1) FTASGA(1) VSDIDQ(1) ALPDTL(1) ITTHMH(1) IYYFLS(1) APARMP(1) 13 Phage display (common panning) 3 PMID:10967104 PLCγ1-C-SH2 domain Not determined. Not determined. Not determined. f3-6mer phage display library (X6) The TrpE-PLC The sequence GDYTLF is selected by each of the SH2 domains of GAP. 750 GDYTLF(4) FAQVWG(3) GSGSWG(1) SFVAYA(1) YVNADA(1) HWFLRH(1) VGWHK(1) GITPNH(1) DSWYRD(1) YPAHGL(1) FSFRFD(1) 11 Phage display (common panning) 3 PMID:10967104 GAP-N-SH2 domain Not determined. Not determined. Not determined. f3-6mer phage display library (X6) The TrpE-GAP-N-SH2 fusion proteins immobilized using anti-TrpE antibodies and protein A-Sepharose beads. The sequence GDYTLF is selected by each of the SH2 domains of PLCγ1. 751 WYGRRW(6) GDYTLF(4) KRRQEP(3) RYFTYP(2) ACDRYV(1) AMDLFT(1) VQYVWT(1) VKNSGP(1) 8 Phage display (common panning) 3 PMID:10967104 GAP-C-SH2 domain Not determined. Not determined. Not determined. f3-6mer phage display library (X6) The TrpE-GAP-C-SH2 fusion proteins immobilized using anti-TrpE antibodies and protein A-Sepharose beads. The sequence GDYTLF is selected by each of the SH2 domains of PLC 752 GDYTLF(5) RILRFY(4) GSGSWG(1) SAVRII(1) SAVNVI(1) SATSRS(1) REAYVF(1) FAAALI(1) KYMPCG(1) ETELTY(1) HGVGSV(1) GVAVTT(1) GLVSNL(1) 13 Phage display (common panning) 3 PMID:10967104 GAP-N+C-SH2 (includes SH3 domain) Not determined. Not determined. Not determined. f3-6mer phage display library (X6) The TrpE-GAP-SH2 fusion proteins immobilized using anti-TrpE antibodies and protein A-Sepharose beads. The sequence GDYTLF is selected by each of the SH2 domains of PLC 753 RGDLKQLSELTW RGDLAALSAPPV SHARGDLVGLTP ILARGDLSTLSA NARGDLGTLPPR RGDLPHQYQHPS RGDLQYPRSLTL RGDLMLPFWPRL RGDLAAPHRTGA RGDLNNYLLMML RGDLPPVPMARE RGDLPQERHYPR RGDLLHASARAS RGDAPSPNIFRL VAPRGDRPTLPI RGDYYANLANWL HARGDFSWLMPE RGDFAQLLITWQ RGDIPSMSTLPR RTDLDSLRTYTL GDLDLLKLRLTR GDLHSLRQLLSR RTDLHTLSSTRH RDDLHMLRLQLW SSDLHALKKRYG TSDLETLRLRLH RDLETLYNRYAT RHDLSELIRRTT HPRTDLASLAKR QRSDLASLKLTL VTDLQSLYWRIK RDDLQTLLNRLG RSDLDTLLARHQ QSGDLYTLRNSR RSDLTALTQLLL KGDLATLIRTNL QSAHRGDIPNVL ASDISALSARLR RDFQYLWNRVSQ VSDVYQLTERLR AGDLVTFKLRLH HHELDSLRARLM RGD, D-L-x(2)-L 42 Phage display (common panning) 3 PMID:9890954 Integrin alpha-V-beta-6, integrin αVβ6 Fibronectin, FN Not determined. Not determined. Ph.D.-12 phage display library (X12) RTDLDSLRTYTL, selected from the phage sequences was a selective inhibitor of RGD-dependent ligand binding to αvβ6 in isolated receptor assays (IC50 = 20 nM), and in cell adhesion assays (IC50 = 50 mM). DLXXL peptides were highly specific inhibitors of αvβ6-fibronectin interaction as synthetic scrambled or reversed DLXXL peptides were inactive. 754 RTDLSML RGDLHSL RGDLYTL RGDLLHT RGDLTSQ RGDLPPS RGDLPPY RGDLPHS RGDLPSP RGDLYVG RGDIPPL RGDILPR RGDMQMP RGDMLTH RGDHLSH RGDAYPS RGDAMPP RGDTSAL LLRGDMS LLRGDMA RGD 20 Phage display (common panning) 3 PMID:9890954 Integrin alpha-V-beta-6, integrin αVβ6 Fibronectin, FN Not determined. Not determined. Ph.D.-7 phage display library (X7) 755 RGDALTILPPML RGDIYFHLAPRN RGDLDNYPGATM RGDSMGPFTVKM RGDSFDTRYAPL ARGDGLLFHLSG TRGDHFMPPWWG VRGDALPDPAIG IPRGDHYLWPQI AMRGDTYLHPTK WLQRGDGFLQLP YGPARGDHFRWG AFNIRGDHWVPD TEISPRGDNFIN NTHAPRGDGYPA GIVTCRGDLFCS DGRSYSTKYWFM NGRLDHPSSPGQ GRIFHTIGPSER GRMPFPNFSPHG GRFPFIASALPP RGD 21 Phage display (common panning) 3 PMID:9890954 Integrin alpha-V-beta-3, integrin αvβ3 Fibronectin, FN Not determined. Not determined. Ph.D.-12 phage display library (X12) 756 GAHWQFNALTVR(13)[1.15 ± 0.13] TSYGRPALLPAA(1)[0.36 ± 0.31] MDHLAPTFRPAI(1)[0.27 ± 0.27] TLRAIWPMWMSS(1)[0.22 ± 0.12] IPLTANYQGDFT(1)[NT] 5 Phage display (common panning) 4 PMID:10993908 Hyaluronan, HA Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay Uncoated paramagnetic beads (Dynal) were labeled with 2 mg/ml HA. To test binding, 125I labeled peptides in 3% BSA in PBS were incubated with the HA-coated beads, and then counted for radioactivities. HA-specific binding was determined by subtracting the baseline cpm that bound to noncoated beads from the cpm associated with the HA-coated beads. Specific binding was then calculated from the specific activity (~3.45e7 cpm/μg) and the number of beads added to the assay. Data shown were reproduced from the graph. NT denotes not tested. GAHWQFNALTVR showed specific binding to soluble, immobilized, and cell-associated forms of HA, and it inhibited leukocyte adhesion to HA substrates almost completely. 757 LQAKKKRPK(3) VIAKIKKPK(3) VEAKGHKKK(2) KWKTPKVRV(1) AIAKKHKWN(1) KWKLHGHIK(1) GKGPKWMR(1) 7 Phage display (common panning) 3 PMID:11070036 Transferrin receptor protein 1 Iron-loaded human transferrin Not determined. Not determined. X9 phage display library Bound phage were eluted with low-acid buffer (0.1 M glycine, pH 2.2). 758 GHKAKGPRK(3) KDKIKMDKK(1) 2 Phage display (competitive panning) 3 PMID:11070036 Transferrin receptor protein 1 Iron-loaded human transferrin Not determined. Not determined. X9 phage display library Bound phage were eluted with ligand (iron-loaded human transferrin; Sigma-Aldrich) in TBS buffer (50 mM Tris-Cl, pH 7.5; 150 mM NaCl). 759 IEAYAKKRK(2)[1.78] GHKVKRPKG(2)[1.94] KNKIPKSPK(1)[NT] 3 Phage display (competitive panning) 3 PMID:11070036 Transferrin receptor protein 1 Iron-loaded human transferrin Not determined. Not determined. X9 phage display library ELISA Absorbance at 490 nm was measured. Data shown were repeoduced from the graph and expressed as the means ± SEM. NT denotes not tested. Bound phage were eluted with low-acid buffer (0.1 M glycine, pH 2.2) and ligand (iron-loaded human transferrin; Sigma-Aldrich) in TBS buffer (50 mM Tris-Cl, pH 7.5; 150 mM NaCl). 760 FDDARL FSDARL FSDMRL FVDVRL FTDIRL FNDYRL FSDTRL 7 Phage display (common panning) 3/4 PMID:10583225 M protein from patient 3 Not determined. Not determined. Not determined. f3-6mer phage display library (X6) 761 DPAFIFYHSTLFFNS GGHDGDPVLTGTLFY AVDPRMFYLLLRGGA 3 Phage display (common panning) 3/4 PMID:10583225 M protein from patient 3 Not determined. Not determined. Not determined. X15 phage display library 762 PIHYIF YIHYIF RIHYIF WREWFL WWAMKP 5 Phage display (common panning) 3/4 PMID:10583225 M protein from patient 6 Not determined. Not determined. Not determined. f3-6mer phage display library (X6) 763 LILSSGELLRHPRGA 1 Phage display (common panning) 3/4 PMID:10583225 M protein from patient 6 Not determined. Not determined. Not determined. X15 phage display library 764 QGAAPFS(1) SLRDTPL(1) LPSYQAI(1) QNPHSPR(1) LNYHRPS(1) LTNHGTA(1) 6 Phage display (common panning) 1 PMID:11839467 Hhydroxyethyl radical-modified bovine serum albumin, BSA-HER Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 765 FGNPGAA(1) WAMSPTK(1) NIDNPNK(1) FHENWPS(1) GQLPSSP(1) YNLPWLN(1) 6 Phage display (common panning) 2 PMID:11839467 Hhydroxyethyl radical-modified bovine serum albumin, BSA-HER Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 766 FHENWPS(15) 1 Phage display (common panning) 3 PMID:11839467 Hhydroxyethyl radical-modified bovine serum albumin, BSA-HER Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 767 SPRPPYP(1) SLMANLI(1) LQPTYSL(1) IPSIPTM(1) 4 Phage display (common panning) 1 PMID:11839467 Acetaldehyde-modified bovine serum albumin, BSA-AcH Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 768 NMERVPV(2) ASFAYTT(1) TPTTTRS(1) WPTMLSK(1) SLPPNWP(1) 5 Phage display (common panning) 2 PMID:11839467 Acetaldehyde-modified bovine serum albumin, BSA-AcH Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 769 GPRAYHE(2) YGTTASL(2) LRHPTLP(1) IGLPLRS(1) WTWPPAY(1) SPDSRLR(1) AAPMALF(1) LPAPPFR(1) TPQTWPL(1) DVELRPL(1) TNNKPIF(1) 11 Phage display (common panning) 3 PMID:11839467 Acetaldehyde-modified bovine serum albumin, BSA-AcH Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 770 IMFESN(2)[ND, ND] IMFQSA(1)[ND, ND] NYFVQG(1)[ND, ND] NYFVET(1)[ND, ND] LFTEYG(1)[0.0027 ± 0.0007, 0.00090 ± 0.00007] NYLVTS(1)[0.036 ± 0.008, 0.024 ± 0.006] NMLVYS(1)[0.16 ± 0.03, 0.032 ± 0.003] VYHVST(1)[0.031 ± 0.003, 0.04 ± 0.01] VFVETS(1)[0.9 ± 0.2, 0.10 ± 0.02] IFLETS(1)[1.3 ± 0.3, 0.2 ± 0.1] VYLATD(1)[0.028 ± 0.005, ND] VFVVNG(1)[0.03 ± 0.01, ND] VFTEER(1)[NT, NT] LTMVQE(1)[NT, NT] QDFPMY(1)[NT, NT] VMFQTD(1)[NT, NT] SYVEYQ(1)[NT, NT] LVAFAN(1)[NT, NT] VHFISR(1)[NT, NT] NSVMIA(1)[NT, NT] FIGVSY(1)[NT, NT] VHVEYT(1)[NT, NT] MWFEAP(1)[NT, NT] VFYSRE(1)[NT, NT] EMFFPV(1)[NT, NT] AFMTRG(1)[NT, NT] IYLVEN(1)[NT, NT] VFVEMP(1)[NT, NT] LYYVTE(1)[NT, NT] WVMTEA(1)[NT, NT] IYYSVS(1)[NT, NT] LMEVSK(1)[NT, NT] VLFVSS(1)[NT, NT] YLEVSS(1)[NT, NT] YVVVSN(1)[NT, NT] NMVMFG(1)[NT, NT] GLNMPA(1)[NT, NT] NFAAFS(1)[NT, NT] FAMAEA(1)[NT, NT] INFESG(1)[NT, NT] LVLQEG(1)[NT, NT] VMPTMS(1)[NT, NT] AYLIQG(1)[NT, NT] TFQVQ(1)[NT, NT] 44 Phage display (common panning) 1-5 PMID:10964781 HIV-1 protease, HIV-1 PR Not determined. Not determined. Not determined. fTC-LIB-N6 phage display library Binding assay The kcat (1/s) and Km (μM) values of peptides with sufficient solubility to conduct kinetic measurements were determined at the pH used in the phage selections, pH 6.7, and additionally at pH 5.6, which is closer to the pH used in many substrate and inhibitor studies of HIV-1 protease. Data were shown as Kcat/Km (1/μMs). ND represents not determined, NT not tested. 771 PIGLRS(4) RPYGLL(3) RPAPLM(2) PALRSR(1) PAGIYA(1) HPLPLS(1) LPRPML(1) PTPAVT(1) GPRPLK(1) PSPDPL(1) PVPLPL(1) KTLPQA(1) SPYPLR(1) KPFLRG(1) PRPFLR(1) RALPPS(1) GPGPGL(1) PRALRA(1) PLPLVR(1) P-x-[GP]-L 19 Phage display (common panning) 4 PMID:10600499 Matrix metalloproteinase-14, MMP-14 proMMP-2 Not determined. Not determined. fUSE-His-N6 phage display library At round 4 selection, the phage were initially immobilized to Ni-NTA and substrate phage were released by proteolytic cleavage. This technique was adopted to prevent an accumulation of "nonreactive" phage, which do not interact with Ni-NTA even before treatment with protease. 772 SRLPSCKRSYCSVAH(6) SRLVGYCTRSPAVCR(5) TFTRVVTDVYRGRLS(3) TWPVVHGACRAHGHC(2) TLIPRSFCPTHDRDC(2) VSLFYTSHVAVRCCN(2) VRLPGKLLRSCERGF(1) RASWLLAFVKRAPFS(1) SAVGFWKSRLAGAYF(1) TRVMFDRGITHHCCH(1) TFRVVCSLPRAGCTAH(1) LCMVRNLLTSPLHHA(1) VWWRALLVFATLYSV(1) MSVWVTHNRVESWFA(1) SSIRISYTARFSGFL(1) 15 Phage display (common panning) 4 PMID:9761687 Protein Rev Not determined. Not determined. Not determined. f3-15mer phage display library (X15) 773 SRLPSCKRSYCSVAH(17) TLIPRSFCPTHDRDC(4) WRISGGLVYFMRYAS(2) SRLVGYCTRSPAVCR(2) TFTRVVTDVYRGRLS(2) SRLPSYNHFFILGSH(1) SHLVRGSFGAGVLVR(1) TRVMFDRGITHHCCH(1) 8 Phage display (common panning) 5 PMID:9761687 Protein Rev Not determined. Not determined. Not determined. f3-15mer phage display library (X15) 774 FLPRGDDYPRR[1.66] VQGRGDSHPAI[1.37] HPCRGDCLA[1.02] KPDRGDYAP[0.91] TTARGDSDK[1.57] ILDRGDSYY[NT] 6 Phage display (common panning) 3 PMID:10869561 Integrin alpha-V-beta-3, integrin αvβ3 Cytotoxic T-lymphocyte protein 4 Not determined. Not determined. CTLA-4 (X3-RGD-X3) scaffold library ELISA Absorbance at 450 nm was measured. Data shown were reproduced from the graph. NT denotes not tested. 775 CPWWDDPQDIC CRWWQETDDSC CEADDSSPGRC CNIYAELDDSC CEYSDSGIEVC CALSENKLRVC CAQSENARPVC CHSSENRSSC CFPSAFSENQC CLPFSENRTC CPTSENRAHSC CLPSPLLENRC 12 Phage display (common panning) 3 PMID:9671121 Anti-BLG polyclonal antibody Beta-lactoglobulin, Beta-LG Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) 776 CEYSDSGIEVC CALSENKLRVC CHSSENRSSC CPTSENRAHSC CLPSPLLENRC CSLYENRRVC CIKNPSSENAC CFTPEGDHRPC CQFFPEGDYLC CSDIDLLSQVC CRQDLSTLDC CRASVDRDLAC 12 Phage display (common panning) 3 PMID:9671121 Anti-BLG polyclonal antibody Beta-lactoglobulin, Beta-LG Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) 777 YSPFSW[37, 110] YVSWSPD[35, 100] FSWSDT[19, 90] YSWSDM[10, 60] WSPFPS[26, 60] DSPFSF[22, 80] FSPFDW[21, 90] TSPFPW[31, 90] YSFFPW[16, 70] YSDFPW[26, 60] DSWFPW[14, 20] SSFYSS[22, 90] DSWSTS[NT] FSPFSF[NT] SSPFDW[NT] 15 Phage display (common panning) 8 PMID:10547288 Protein Nef Tyrosine-protein kinase HCK Not determined. Not determined. RRT-SH3 phage display library Binding assay The relative potency of the corresponding SH3 domains to compete with labeled Hck-SH3 Nef and RRT.A1 Nef in binding to Nef, respectively, expressed as probe:competitor ratio at 50% inhibition. NT denotes not tested. To examine if it would be possible to present functional SH3 domains on the surface of bacteriophages authors constructed a phagemid containing Hck-SH3, and by using a M13KO7 helper virus produced recombinant phages expressing it fused to the pIII coat protein. 778 QGFLDQ[60] NAFLPS[40] EAWSPL[80] ESYSEW[10] 4 Phage display (common panning) 8 PMID:10547288 Protein NefR90 Tyrosine-protein kinase HCK Not determined. Not determined. RRT-SH3 phage display library Binding assay The relative potency of the corresponding SH3 domains to compete with labeled RRT.A1 NefR90 in binding to NefR90, expressed as probe:competitor ratio at 50% inhibition. To examine if it would be possible to present functional SH3 domains on the surface of bacteriophages authors constructed a phagemid containing Hck-SH3, and by using a M13KO7 helper virus produced recombinant phages expressing it fused to the pIII coat protein. 779 SPTRAVNELFGM(11)[0.21] SLPRVYYEIFGA(6)[0.40] HTNLPQALWGMQ(5)[0.10] KLAQTFPQVLLA(5)[NT] HNNQRLMLYGMH(3)[0.05] HWPFLQWHMYYP(3)[NT] YTAAPRYMAELF(2)[NT] YVGPLTQTLYGM(2)[0.08] GHPRTAELLGML(1)[0.19] LRPFHQILYGMS(1)[0.14] TLNWPPTLWGMH(1)[0.05] YFWPMTKFQLWR(1)[0.05] TASTLHKTLFGM(1)[NT] FYGLHASLFGMR(1)[NT] HWSHAFMSVWGM(1)[NT] QMLNRLSELNGM(1)[NT] SLVELPSPPMLA(1)[NT] TFLANPSSNLVM(1)[NT] SFAASEPDRART(1)[NT] TSFTGWWNLMAR(1)[NT] GPEQLLFLRPFP(1)[NT] APMHKALWVQPG(1)[NT] GQLQFSNRSLSV(1)[NT] MYNDRTRSFIAS(1)[NT] FNSSHSLLRRPP(1)[NT] QIRTLPIYSLQS(1)[NT] YHWWQFQMKHSA(1)[NT] YRWWPFQVKPSA(1)[NT] WWHPKIPSPSAR(1)[NT] WWQFPWPPYTSV(1)[NT] NSVIGCRTQSCD(1)[NT] NQLIASLSPRVD(1)[NT] RFPWFFMESSHS(1)[NT] 33 Phage display (common panning) 4 PMID:11275260 Anti-NTX monoclonal antibody BNTX18 Potassium channel toxin alpha-KTx 2.1 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance at 405 nm was measured. Each value represents mean of triplicates. Data shown were reproduced from the graph. NT denotes not tested. We used the biopanning procedure in two conditions: with the mAb immobilized on a plastic surface (three rounds) and in solution (the final, fourth, round). In total authors determined the sequences of 33 different peptides displayed by 59 clones isolated from the eluates after the third and the fourth rounds of panning of the 12-mer peptide library. 780 MVLWGMH(7) QWIFGMS(1) EQWLGMS(1) EALYGMS(1) QWLYGMS(1) AIAPFFA(1) PHLPMQS(1) SIASLSR(1) SDSSTLN(1) HTTQHRL(1) 10 Phage display (common panning) 3 PMID:11275260 Anti-NTX monoclonal antibody BNTX18 Potassium channel toxin alpha-KTx 2.1 Not determined. Not determined. Ph.D.-7 phage display library (X7) In total authors determined the sequences of 16 clones containing ten different peptides derived from the 7mer library. 781 CIVPKPASEQC[0.91] CLVPRRPPPQC[0.66] CVFLTAQTVPC[0.78] CVAYETAIASC[0.67] CFIPPINPATC[0.64] CRPPLRTKKSC[0.59] CGPAAARLDMC[0.54] CFVPAVQPATC[0.84] CIISPRSVSPC[1.05] CLVPAPIIALC[0.61] 10 Phage display (common panning) 3 PMID:10381087 Cucumber mosaic virus, CMV Not determined. Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA Absorbance at 405 nm was measured. Background signals of 0.11, obtained with M13KO7 helper phage, were subtracted from signals produced with peptide displaying phage. Data shown were reproduced from the graph. CMV was also isolated in house from 50g of infected Nicotiana cle6elandii leaf tissue. Sequencing of the genes encoding 10 of these peptides revealed an absence of any conserved motifs, although nine of them contained a high proportion of proline residues. 782 CVFTSDYAFC(2)[7.8] CVIYDGNHWC(2)[NT] CIFEPDYSYC(2)[NT] CVFDDLYSFC(1)[NT] 4 Phage display (common panning) 3 PMID:11012675 Prostate-specific antigen Not determined. Not determined. Not determined. CX8C phage display library Surface plasmon resonance (SPR) Equilibrium dissociation constant (KD, μM) was measured. The values shown are the average KD of three different concentrations of analyte (GST-peptides). PSA was immobilized on microtiter wells coated with the monoclonal IgG 5E4, which binds both free and complexed PSA. 783 CTFSVDYKYLMC(15)[NT] CVFAHNYDYLVC(2)[3.5] CRFDKEYRTLVC(1)[NT] 3 Phage display (common panning) 3 PMID:11012675 Prostate-specific antigen Not determined. Not determined. Not determined. CX10C phage display library Surface plasmon resonance (SPR) Equilibrium dissociation constant (KD, μM) was measured. The values shown are the average KD of three different concentrations of analyte (GST-peptides). PSA was immobilized on microtiter wells coated with the monoclonal IgG 5E4, which binds both free and complexed PSA. 784 CLSTCAQSCRISC(7) CLLYCHDACWWVC(2) 2 Phage display (common panning) 3 PMID:11012675 Prostate-specific antigen Not determined. Not determined. Not determined. CX3CX3CX3C phage display library PSA was immobilized on microtiter wells coated with the monoclonal IgG 5E4, which binds both free and complexed PSA. 785 CVEYCWEGSCYVC(7)[NT] CVAYCIEHHCWTC(3)[2.9] CVSYCLFEFCYVC(2)[NT] CVAYCEEWECYVC(1)[NT] CVSYCDGLQCWMC(1)[NT] 5 Phage display (common panning) 3 PMID:11012675 Prostate-specific antigen Not determined. Not determined. Not determined. CX3CX4CX2C phage display library Surface plasmon resonance (SPR) Equilibrium dissociation constant (KD, μM) was measured. The values shown are the average KD of three different concentrations of analyte (GST-peptides). PSA was immobilized on microtiter wells coated with the monoclonal IgG 5E4, which binds both free and complexed PSA. 786 DPGVEVWLGN(5) GGTTVWLGEG(3) GTEVWLWGGG(3) RGITVVLGKS(2) WGTEVWLGMG(1) RGVTVWLGGT(1) RGVTVWLGQF(1) LVGTDIWLWT(1) G-x(2)-V-W-L-G 8 Phage display (common panning) 3 PMID:9894603 PDZ2 domain of hPTPE1 Not determined. Not determined. Not determined. PDL-10R phage display library PDZ2 expressed as a fusion protein (GST-PDZ2) was immobilized in wells of microplates, and affinity panning was done. 787 KAYGTVVWLG(2) NSYGTVVWLG(1) VVYGTVVWLG(1) DSYGIEVWLG(1) WEYGIEVWLG(1) G-x(2)-V-W-L-G 5 Phage display (common panning) 3 PMID:9894603 PDZ2 domain of hPTPE1 Not determined. Not determined. Not determined. PDL-10Y phage display library PDZ2 expressed as a fusion protein (GST-PDZ2) was immobilized in wells of microplates, and affinity panning was done. 788 QKRAKGLSA(8)[0.441 ± 0.060] YRKKSSAEL(2)[0.496/0.363] MRKANSPPT(2)[0.491/0.401] ARKVKGAAG(2)[0.404/0.384] RGLKSKIAM(1)[0.466] FRKTSISGA(1)[0.421] LSRKKTLTT(1)[0.421] RSLKTKLPS(1)[0.402] LRKKGYDPG(1)[0.400] SRKKTFTGA(1)[0.400] TKLSNTPSP(1)[0.396] GRKSAVSPA(1)[0.358] HRKSTVAGS(1)[0.345] FRKTSKGGS(1)[0.341] HKKSLSSPS(1)[0.338] RRKPGTEQL(1)[0.332] YRKTAKEGT(1)[0.323] YKKGARPIQ(1)[0.323] RK 18 Phage display (subtractive panning) 3/5 PMID:11034389 Anti-GAD65 monoclonal antibody MICA3 Glutamate decarboxylase 2 Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA The reactivity of phagotopes expressing that peptide with MICA3, as determined by capture ELISA, is given as an OD value measured at 405 nm. After each round of positive selection, there was a negative selection step in which phage not specifically reactive with the mAbs were removed using magnetic beads coated with anti-human Ig in the absence of primary Ab. Twenty-eight of 35 phagotopes from the fifth round of positive selection and 0 of 32 phage from the third round of positive selection were reactive by capture ELISA. 789 MRKSTGTAS(2)[0.461/0.473] QKKMVALSG(1)[0.733] SRKVALQGG(1)[0.639] RRKLRTNAG(1)[0.631] RKRTRHLEG(1)[0.562] TQDLGYSRV(1)[0.561] RKKKPPGLA(1)[0.554] TRKALTQNT(1)[0.541] NRKLKSNMM(1)[0.523] KRRASAGGP(1)[0.491] RKVVQTSLE(1)[0.476] KGKKALQSA(1)[0.467] VRKLTVTSA(1)[0.446] KKTKGLVTT(1)[0.435] KSVKPRQAT(1)[0.434] FRKADKFPM(1)[0.434] RKTALGTRQ(1)[0.426] SRKSGSLTK(1)[0.425] KKKKYAEAV(1)[0.400] ARLSVVRNG(1)[0.370] SRKATASLP(1)[0.367] TRGLKGAPQ(1)[0.320] RKLAHPGTS(1)[0.308] GGTTTPRTT(1)[0.294] NRRLLPMPE(1)[0.290] V-A-L-x-G 25 Phage display (subtractive panning) 3/5 PMID:11034389 Anti-GAD65 monoclonal antibody MICA4 Glutamate decarboxylase 2 Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA The reactivity of phagotopes expressing that peptide with MICA4, as determined by capture ELISA, is given as an OD value measured at 405 nm. After each round of positive selection, there was a negative selection step in which phage not specifically reactive with the mAbs were removed using magnetic beads coated with anti-human Ig in the absence of primary Ab. Twelve of 14 phage from the fifth round of positive selection and 14 of 21 phage from the third round of positive selection reacted by capture ELISA. 790 WDPAMNVSLQNS(4) YDPAQGRIAGIR(1) TDPAHTPFPHTR(1) WDPAQQVRHQDN(1) SDPAQAVGRMRP(1) GGSGTSRTPILG(1) IHLPIASASQMT(1) 7 Phage display (common panning) 1 PMID:11602054 Anti-E1 protein polyclonal antibody IgG HCV E1 protein Not determined. Not determined. Ph.D.-12 phage display library (X12) 791 NLIGHHFPHFRALSS TWPVVHGACRAHGHC GGFSLHPWWRFYHDR THSHQWRHHQFPAPT QHGHFTDGYHLPSRL GFTDVHLHLPGNSHR LTLSHPHWVLNHFVS 7 Phage display (common panning) 4 PMID:12174288 HrNMT Not determined. Not determined. Not determined. f3-15mer phage display library (X15) 792 TWPVVHGACRAHGHC NLIGHHFPHFRALSS GLDLLGAVRKPVVRR GALVLSRVDRWFLVA TASLVFFRDGTLSNR LVRHGRYLVTAWHSY AFVALGSLSYGLHGP FAFGFFGGRVWIPRG EFLHLSPTGGVWLAS AGPLRGFYMCSWCTP 10 Phage display (common panning) 4 PMID:12174288 Glycylpeptide N-tetradecanoyltransferase Not determined. Not determined. Not determined. f3-15mer phage display library (X15) 793 PHCVPRDLSWLDLEANMCLP(18) VTCGSIAEYGWLDLAAACSS(1) PRCMQTSYWMDGLQPESCKG(1) RVCAAPESRLFRGMPLGCDD(1) 4 Phage display (common panning) 3 PMID:10493820 14-3-3 protein theta Not determined. Not determined. Not determined. X2CX14CX2 phage display library In fact, microtiter wells were coated overnight with GST-14-3-3τ in TBS.', '3', '10493820', 'PHCVPRDLSWLDLEANMCLP efficiently blocked the binding of 14-3-3 to the kinase Raf-1, a physiological ligand of 14-3-3, and effectively abolished the protective role of 14-3-3 against phosphatase-induced inactivation of Raf-1. PHCVPRDLSWLDLEANMCLP efficiently blocked the binding of 14-3-3 to the kinase Raf-1, a physiological ligand of 14-3-3, and effectively abolished the protective role of 14-3-3 against phosphatase-induced inactivation of Raf-1. 794 RNCWGNIPLTSSSVERLCDAR(1) DACSKQGMGVLLSGWPGPCTT(1) PACLLRSEEYVVECGGDVGLE(1) VCCGVNESLSRSAHADSALMR(1) EVCHMPVSCGPTERSLGGESL(1) 5 Phage display (common panning) 3 PMID:10493820 14-3-3 protein theta Not determined. Not determined. Not determined. X2CX18 phage display library In fact, microtiter wells were coated overnight with GST-14-3-3τ in TBS. 795 NCPLNYTYTYRQ HPPDYNVIYRTP DSPLNYTYTYRQ EAPLQYSYSYRA YQPFNYSVLYRA APQTYQTSYRSP MHWPVDYSSTYR RFPVSYTQSYRT WLHPLSYTEAYR VNQPVDYSSTYR SNRPVEYTHVYR APVSYDSHYRGR VNQPSNYTQIYR SNRPLDYSHVYR YSPVTYTKYRLP 15 Phage display (common panning) 3 PMID:10353843 Anti-human SP-A monoclonal antibody PE10 Pulmonary surfactant-associated protein A1 Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage selected by mAbs displayed the consensus peptide sequences that are nearly identical to 184TPVNYTNWYRG194 of human SP-A. 796 SFPPDYWRHYRM CQIPHSYGGYRS VLPHTYSQYRL SALSYTTVYRSP HQQQQDNSSYRQ GSTPQHYTQLYR SPIPHSYGGHLR YRSPPNRWIQYR QNPESFHEYRTN IPHAPRSYVTRY TGPLHYSSQNRH YYPLDYSRHYRM YPSPPNRLIQYR DHPPWYTWLYR TGTPPFQCASYR NQEPGNYASTYR 16 Phage display (common panning) 3 PMID:10353843 Anti-human SP-A monoclonal antibody PC6 Pulmonary surfactant-associated protein A1 Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage selected by mAbs displayed the consensus peptide sequences that are nearly identical to 184TPVNYTNWYRG194 of human SP-A. 797 SDLVSSRML(2) ELFSTKMDI(2) DDELHTSRY(2) EDLETSIWR(1) RDLLTMRML(1) NDLLTSRVL(1) MDLYSQRIL(1) YWDLTSSKL(1) MMDLDTEYL(1) GTDLESYKL(1) ELKTTKYQD(1) EELSTSKWL(1) IDMVMSKMV(1) SLQTFKLMP(1) ETLTTSKWR(1) SNLETSRFY(1) EGLDTWPMR(1) GTEIMTQYL(1) RTSLETWAI(1) GFLDAWLGE(1) 20 Phage display (common panning) 3 PMID:10029604 Human neutrophil cells Not determined. Not determined. Not determined. J404 phage display library (X9) Freshly isolated PMN were incubated with 20 μL aliquots of phage library in 1 mL HBSS containing 0.1% bovine serum albumin (phage buffer) for 1 hour at 20°C in a 1.5 mL microcentrifuge tube. PMN were then washed five times with phage buffer and bound phage were eluted for 5 minutes in 2 mL of 0.1 mol/L glycine (pH 2.2). 798 DLETSFMKV(3)[NT] DLVTSKLQV(1)[0.76 ± 0.02] DLETSFMPA(1)[NT] DLQTEKYMG(1)[NT] DLMTSKYMA(1)[NT] DLETKFMRQ(1)[NT] KDLLTTKLS(1)[NT] KDLQTRAMH(1)[NT] KDLATSKLL(1)[NT] RDLSTRPLW(1)[NT] RDLLTSKYV(1)[NT] TDLQTMAMR(1)[NT] NLDLHTEKI(1)[NT] RAGDLTTTK(1)[NT] ELNTWKMDW(1)[NT] DPESSFMKV(1)[NT] NPRLETYAM(1)[NT] RALSPPPMS(1)[NT] SSRLETSYL(1)[NT] ENLSNWQIS(1)[NT] 20 Phage display (common panning) 3 PMID:10029604 Human neutrophil cells Not determined. Not determined. Not determined. J404 phage display library (X9) ELISA Purified phage clones were incubated in microtiter plates coated with PMN freshly isolated from peripheral venous blood. Bound phages were detected with polyclonal anti-phage antibody and quantified by light absorbance. Optical density values, OD (405 nm), are representative of one of two experiments showing the mean ± SD of quadruplicate determinations. NT represents not tested. Freshly isolated PMN were incubated with 20 μL aliquots of phage library in 1 mL HBSS containing 0.1% bovine serum albumin (phage buffer) for 2 hours at 4°C in a 1.5 mL microcentrifuge tube. PMN were then washed five times with phage buffer and bound phage were eluted for 5 minutes in 2 mL of 0.1 mol/L glycine (pH 2.2) followed by addition of phage buffer containing 0.5% Tween 20 to the remaining cell pellet. 799 FGPNLTGRW(3)[1.16 ± 0.04] DDLSTTRLP(3)[NT] GTDLESYKL(3)[NT] ADLMTKAMH(1)[NT] TPDLETSRL(1)[NT] SNLATSKLL(1)[NT] QWGLQTYKM(1)[NT] SWELLTEAM(1)[NT] SPSLQTFKM(1)[NT] KMDWTTVYK(1)[NT] GAPNLTGRW(1)[NT] GGPNLTGRW(1)[NT] GWLDDMWKG(1)[NT] TLSPWSVWR(1)[NT] APDWWNIWS(1)[NT] WNRIDAWDS(1)[NT] 16 Phage display (common panning) 3 PMID:10029604 Human neutrophil cells Not determined. Not determined. Not determined. J404 phage display library (X9) ELISA Purified phage clones were incubated in microtiter plates coated with PMN freshly isolated from peripheral venous blood. Bound phages were detected with polyclonal anti-phage antibody and quantified by light absorbance. Optical density values, OD (405 nm), are representative of one of two experiments showing the mean ± SD of quadruplicate determinations. NT represents not tested. Freshly isolated PMN were incubated with 20 μL aliquots of phage library in 1 mL HBSS containing 0.1% bovine serum albumin (phage buffer) for 1 hour at 20°C in a 1.5 mL microcentrifuge tube. PMN were then washed five times with phage buffer and bound phage were eluted for 5 minutes in 2 mL of 0.1 mol/L glycine (pH 2.2). 800 DLVTSKLQV(4)[[0.76 ± 0.02]] NDLLTSKMP(4)[NT] DLSTSKLQI(2)[NT] DLPTSKMNL(2)[NT] DLETSFMKV(2)[NT] ADLAMSKWR(2)[NT] LHFADVWK(2)[NT] DLNTYKMSL(1)[NT] DLAKSKLML(1)[NT] DDLVTQKMA(1)[NT] KDLFSWQLI(1)[NT] KDLVTQKYF(1)[NT] EDLSTQRIK(1)[NT] EMDWMKIWR(1)[NT] 14 Phage display (common panning) 3 PMID:10029604 Human neutrophil cells Not determined. Not determined. Not determined. J404 phage display library (X9) ELISA Purified phage clones were incubated in microtiter plates coated with PMN freshly isolated from peripheral venous blood. Bound phages were detected with polyclonal anti-phage antibody and quantified by light absorbance. Optical density values, OD (405 nm), are representative of one of two experiments showing the mean ± SD of quadruplicate determinations. NT represents not tested. Freshly isolated PMN were incubated with 20 μL aliquots of phage library in 1 mL HBSS containing 0.1% bovine serum albumin (phage buffer) for for 1 hour at 20°C in a 1.5 mL microcentrifuge tube. PMN were then washed five times with phage buffer and bound phage were eluted for 5 minutes in 2 mL of 0.1 mol/L glycine (pH 2.2) followed by addition of phage buffer containing 0.5% Tween 20 to the remaining cell pellet. 801 CLDTAKYFWC(2) CSCAGKMWCRC(2) CADGTSKVRLC(1) CLVSERLC(1) CLSKDLASSPMC(1) CDWGDLESSVLC(1) CLASYRFGDRYC(1) CWMGWKCSDTGC(1) CDPISMLGKWYC(1) CLVEGWCNPWVC(1) 10 Phage display (common panning) 3 PMID:10029604 Human neutrophil cells Not determined. Not determined. Not determined. CL10 phage display library (CX10C) Freshly isolated PMN were incubated with 20 μL aliquots of phage library in 1 mL HBSS containing 0.1% bovine serum albumin (phage buffer) for 2 hours at 4°C in a 1.5 mL microcentrifuge tube. PMN were then washed five times with phage buffer and bound phage were eluted for 5 minutes in 2 mL of 0.1 mol/L glycine (pH 2.2). 802 CNLVTTTKYNPC(9) CSCLVTEKFSSC(1) CEIQTTKMGMAC(1) 3 Phage display (common panning) 3 PMID:10029604 Human neutrophil cells Not determined. Not determined. Not determined. CL10 phage display library (CX10C) Freshly isolated PMN were incubated with 20 μL aliquots of phage library in 1 mL HBSS containing 0.1% bovine serum albumin (phage buffer) for 2 hours at 4°C in a 1.5 mL microcentrifuge tube. PMN were then washed five times with phage buffer and bound phage were eluted for 5 minutes in 2 mL of 0.1 mol/L glycine (pH 2.2) followed by addition of phage buffer containing 0.5% Tween 20 to the remaining cell pellet. 803 CWWDGMCAALKC(3) CWPSWMCDRLQC(2) CLVEGWCNPWVC(2) CRNCGWDRPNIC(2) CPFGPDLQGKWC(1) CRSNMWLGNWLC(1) CSKDSWLGRIYC(1) CLVKDWCGTMRC(1) CLGDWWQRWELC(1) CTEWGGMWNILC(1) CSMWGGMWEMFC(1) CGSWEMWGKEEC(1) CGRWPTWGDPRC(1) CGDPGTWELLWC(1) CRGMWALLNVAC(1) 15 Phage display (common panning) 3 PMID:10029604 Human neutrophil cells Not determined. Not determined. Not determined. CL10 phage display library (CX10C) Freshly isolated PMN were incubated with 20 μL aliquots of phage library in 1 mL HBSS containing 0.1% bovine serum albumin (phage buffer) for 1 hour at 20°C in a 1.5 mL microcentrifuge tube. PMN were then washed five times with phage buffer and bound phage were eluted for 5 minutes in 2 mL of 0.1 mol/L glycine (pH 2.2). 804 CKDGLFLGSWLC(1)[1.46 ± 0.09] CTRSMFLGEWLC(1)[NT] CMAGVFLGALGC(1)[NT] CDVGALLGPWLC(1)[NT] CNLGNFLGSWLC(1)[NT] CNSEGPLGRWYC(1)[NT] CRKGTLLGEYFC(1)[NT] CADGLLLGKWYC(1)[NT] CNSQGILGAWYC(1)[NT] CDPRQWLGQYLC(1)[NT] CARNGFLGNWLC(1)[NT] CGKDSFLGMWLC(1)[NT] CDKKSWLGSWLC(1)[NT] CDRKFFLGKNWC(1)[NT] CRNCGWDRPMIC(1)[NT] CSTCGWNNEMWC(1)[NT] CLVKDWCGTMRC(1)[NT] 17 Phage display (common panning) 3 PMID:10029604 Human neutrophil cells Not determined. Not determined. Not determined. CL10 phage display library (CX10C) ELISA Absorbance at 405 nm was measured. Data were reproduced from the graph and expressed as representative of one of two experiments showing the mean ± SD of quadruplicate determinations. NT represents not tested. Freshly isolated PMN were incubated with 20 μL aliquots of phage library in 1 mL HBSS containing 0.1% bovine serum albumin (phage buffer) for for 1 hour at 20°C in a 1.5 mL microcentrifuge tube. PMN were then washed five times with phage buffer and bound phage were eluted for 5 minutes in 2 mL of 0.1 mol/L glycine (pH 2.2) followed by addition of phage buffer containing 0.5% Tween 20 to the remaining cell pellet. 805 CQQFLSVRALC CGGTPPSLPTC CGGTRRPIASC CFPYCYPSESA CSEYREPLLC 5 Phage display (common panning) 4 PMID:11053238 Anti-Bet v 1 polyclonal antibody IgE Major pollen allergen Bet v 1-A Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) 806 AGTHTPGAT TSKASELAV 2 Phage display (common panning) 4 PMID:11053238 Anti-Bet v 1 polyclonal antibody IgE Major pollen allergen Bet v 1-A Not determined. Not determined. pVIII-9aa phage display library (X9) 807 AAHMQRFPLLHQ(4) FSKPSAALTHDT(3) MSPMHKQSRATY(3) SFLTPWFTHFPR(2) LATTKPSHLTRL(1) SIHHWPMLFRLP(1) LSPFPKYPPKIP(1) VSHLPTLKMPSA(1) 8 Phage display (common panning) 4 PMID:10936026 Anti-gpIIIa monoclonal antibody Y2/51 Integrin beta-3 Not determined. Not determined. Ph.D.-12 phage display library (X12) 808 HTYSWPTLHPNG(2) MTFSTKGSTTSG(1) STMHHHLAPFIA(1) YVHSTQPLTPQN(1) TPVMPPRWAVAL(1) SSHVQSWRLPKA(1) QGLPFHAALRLL(1) IRPWIWTPDIRS(1) YNTTTPVPQDLS(1) IPLHWHADQTRW(1) LPSSIRLHSHFP(1) FPFQHMSLKHID(1) FSTSPSYSPVWR(1) SWHVWPYHQDRS(1) TPVSNDDLAQKR(1) 15 Phage display (common panning) 4 PMID:10936026 Anti-gpIIb/IIIa monoclonal antibody 5B12 Integrin alpha-IIb/beta-3 Not determined. Not determined. Ph.D.-12 phage display library (X12) 809 LAHPTSGHTLPW(3)[2.22, 1.53] SGLAKFCLAPLP(2)[NT, NT] DVLMHGFDPALI(1)[0.70, 1.53] WRILASGHTLPW(1)[NT, 1.17] QIWHHYPNSQLL(1)[NT, NT] LEMFPDKNQRLT(1)[NT, NT] AKTMVIFLRRRL(1)[NT, NT] YLQSNTEPKQDS(1)[NT, NT] MFPMGMSQRPFS(1)[NT, NT] NRTPEILHSIMH(1)[NT, NT] SVRSPHLFPVMR(1)[NT, NT] TTHHYGYKSGY(1)[NT, NT] 12 Phage display (common panning) 4 PMID:10936026 Anti-human platelet polyclonal antibody Human platelet Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Two types of ELISA were performed. In the first set of experiments all phage clones were evaluated using plates coated directly with phage. In the second set of experiments plates were previously coated with anti-M13 monoclonal antibody, then phage (only phage clones selected with rabbit antihuman platelet antibody) were added. In this case anti-M13 monoclonal antibody reacted with pVIII protein of phage particle, and peptides expressed as fusion with pIII would be accessible for serum IgG. The absorbance was measured at 405 nm, and each OD value represents the mean of duplicate determinations. Platelet lysate was used as positive control. Non-related phage GM6 was used as negative control. Data shown were reproduced from the graph. NT denotes not tested. 810 LASLCNPKPSDAPVT(9) LVRAYAGLARIILKW(4) YASPSLFGFPFALEA(2) ICACLCCACSYNSXV(2) KAHDLSTASRPHVVT(1) PAGEIKVGKLRQLQR(1) SQYGIRGSFLLYVLP(1) 7 Phage display (common panning) 3 PMID:9698107 Anti-chlamydia pneumoniae monoclonal antibody RR-402 Chlamydia pneumoniae Not determined. Not determined. f88-15mer phage display library (X15) LASLCNPKPSDAPVT showed higher ELISA reactivity with C. pneumoniae MIF-positive sera compared to patients with other chlamydial infections, non-chlamydial respiratory infections and normal healthy sera (MIF-negative). 811 GLISDSGFLIPSKRR(2) FVKDRPLELRTFGTL(2) GTVTLGRFSRLSLFG(2) VRVLWGGFSRLSLFX(1) PTTPKSILHFLFPLF(1) LASSVKSAEVGCAGY(1) SVTSATPRGERHVSP(1) QFCGWGEFEVRVLWP(1) NSKGKMNTCHKVGNS(1) 9 Phage display (common panning) 3 PMID:9698107 Polyclonal sera from patients with C. pneumoniae infection Chlamydia pneumoniae Not determined. Not determined. f88-15mer phage display library (X15) LASLCNPKPSDAPVT showed higher ELISA reactivity with C. pneumoniae MIF-positive sera compared to patients with other chlamydial infections, non-chlamydial respiratory infections and normal healthy sera (MIF-negative). 812 VGFALFNARY KYNALYGVRT KGNARYTLLS GWNAQYEWRG KWNGAYRSER VARLNSSYRN N-A-x-Y 6 Phage display (common panning) 3 PMID:9675087 Anti-SHP-1 monoclonal antibody 6D41 Tyrosine-protein phosphatase non-receptor type 6 Not determined. Not determined. PDL-10R phage display library 813 LLYNALYGRE GLYGWNSGYS MKYNAQYRWD PHYNAHYQRG SRYTWNSRYT RMYNALYEAS AWYKLNAGYS N-A-x-Y 7 Phage display (common panning) 3 PMID:9675087 Anti-SHP-1 monoclonal antibody 6D41 Tyrosine-protein phosphatase non-receptor type 6 Not determined. Not determined. PDL-10Y phage display library 814 SKRPLWPVFA PWWPARNRPP MPNWPVLRGP GNPPAWPAPG PAWPDLRGWV DKVTPFWPAL KMPWWPFLEV KLKGPWWPKA PRWPWLEGVG P-x-W-P 9 Phage display (common panning) 3 PMID:9675087 Anti-SHP-1 monoclonal antibody 3E7 Tyrosine-protein phosphatase non-receptor type 6 Not determined. Not determined. PDL-10R phage display library 815 RPYWPIKNLA SPYWPYRMWT KPYWPGGGEA HPYWPDTSRS PYWP 4 Phage display (common panning) 3 PMID:9675087 Anti-SHP-1 monoclonal antibody 3E7 Tyrosine-protein phosphatase non-receptor type 6 Not determined. Not determined. PDL-10Y phage display library 816 GFVWYGLNIK ILDWYGVRID LDWYGILLET RWYDLILLQD WY 4 Phage display (common panning) 3 PMID:9675087 Anti-SHP-1 monoclonal antibody 4E6 Tyrosine-protein phosphatase non-receptor type 6 Not determined. Not determined. PDL-10R phage display library 817 MWYGVEVPWS ELYWYGLPIR YWYGLEIPSA WY 3 Phage display (common panning) 3 PMID:9675087 Anti-SHP-1 monoclonal antibody 4E6 Tyrosine-protein phosphatase non-receptor type 6 Not determined. Not determined. PDL-10Y phage display library 818 KHVQYWTQMFYS(5) DFLQWKLARQKP(1) 2 Phage display (common panning) 4 PMID:9862799 Texas red fluorophore dye Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Each biopanning round consisted of four sequential steps: phage binding with a fluorophore dye carrier, washing unbound phage from the beads, nonspecific elution of bound phage and amplification of bound phage. 819 IPHPPMYWTRVF(6) 6 Phage display (common panning) 4 PMID:9862799 Rhodamine red fluorophore dye Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Each biopanning round consisted of four sequential steps: phage binding with a fluorophore dye carrier, washing unbound phage from the beads, nonspecific elution of bound phage and amplification of bound phage. 820 HGWDYYWDWTAW(4) ASDYWDWEWYYS(2) YPNDFEWWEYYF(2) HTSHISWPPWYF(1) LEPRWGFGWWLK(1) QYYGWYYDH(1) YMYDEYQYWNFW(1) 7 Phage display (common panning) 4 PMID:9862799 Oregon green 514 fluorophore dye Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Each biopanning round consisted of four sequential steps: phage binding with a fluorophore dye carrier, washing unbound phage from the beads, nonspecific elution of bound phage and amplification of bound phage. 821 YPNDFEWWEYYF(7) ASDYWDWEWYYS(2) WYDDWNDWHAWP(2) WHMSPSWGWGYW(1) HMSWWEFYLVPP(1) YWDYSWHYYAPT(1) 6 Phage display (common panning) 4 PMID:9862799 Fluorescein fluorophore dye Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Each biopanning round consisted of four sequential steps: phage binding with a fluorophore dye carrier, washing unbound phage from the beads, nonspecific elution of bound phage and amplification of bound phage. 822 GYARTFWLPGLWSVP(8)[+] WVSHFSVFAFPMSSW(4)[++] AGLHLFWSYSSYVTP(4)[++] SGPFIWNFWLGARAV(3)[+++] RLLVFRYWNLFPNVS(1)[-] GVRVPFWEGLEFFTL(1)[+/-] GSLIVPEWHLAPPGL(1)[-] TNLPPIFYDLHCFAF(1)[+] SGWSAFYNVAFFSTR(1)[++] RNLPPIFNDVYWIAF(1)[-] 10 Phage display (common panning) 4 PMID:10449040 U1 small nuclear RNA, U1 snRNA Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Dot blot Fourth-round clones were initially screened using dot blots. Intensity of each dot was determined subjectively and is indicated as being either strong (+++), moderate (++), weak (+), very weak (+/-), or absent (-) relative to the negative control. 823 AGLHLFWSYSSYVTP(8) FRSWMHFVTYVSTGG(8) EAWARWVFAWLWGHG(6) GYARTFLWPGLWSVP(5) WAYYHAGHSSFAVWT(3) GPQSATSCCYVSTGG(3) YRVSLSVLHNSLLPS(2) VVPRFWFWLHGPGPF(1) VVRAFLGISLGLWSV(1) AHLYFDSGWASRHWT(1) 10 Phage display (common panning) 5 PMID:10449040 U1 small nuclear RNA, U1 snRNA Not determined. Not determined. Not determined. f3-15mer phage display library (X15) 824 SISPWGFSGLLRWSY(28)[+++] TYALTSSILVYLTSS(1)[+] EVPRLSLLAVSLVAN(1)[+] TVSAGIICSFLSVSC(1)[+] VFQFRASVGGSHTVI(1)[-] RLVCWRLGCVSPMGS(1)[+/-] KWPVAVTLAAGIRIP(1)[+/-] SVLSHVRTVVLFSRS(1)[-] LRYTLETRWAVLSVY(1)[++] LILFSYDFHGDYYFY(1)[-] 10 Phage display (common panning) 4 PMID:10449040 Anticodon stem and loop of yeast tRNA(Phe) (tRNA(ACPhe)) Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Dot blot Fourth-round clones were initially screened using dot blots. Intensity of each dot was determined subjectively and is indicated as being either strong (+++), moderate (++), weak (+), very weak (+/-), or absent (-) relative to the negative control. 825 SISPWGFSGLLRWSY(42) AAGGFFRWIFSPVYV(5) VSVYYGSFCLHSAYG(1) NNHPWYYGRSHHVSG(1) VFQFRASVGGSHTVI(1) TFRLLALWRGGLYPG(1) 6 Phage display (common panning) 5 PMID:10449040 Anticodon stem and loop of yeast tRNA(Phe) (tRNA(ACPhe)) Not determined. Not determined. Not determined. f3-15mer phage display library (X15) 826 MDDDTERFPTHRSLP[1.16] XPPFTSAVGGVDHRS[0.45] STTNTPLVSHLEHRS[0.42] LEHRS 3 Phage display (common panning) PMID:10725197 Anti-gG2 monoclonal antibody H5 Envelope glycoprotein G, gG, gG-2 Not determined. Not determined. f3-15mer phage display library (X15) ELISA In ELISA, the wild-type phage f88-4 was used as negative control. Absorbance at 490 nm was measured. Data shown were reproduced from the graph. The A490 of the wild-type phage f88-4 was 0.30. 827 QPTSKPTRPWLVSFL NAETAFSLSSFPPSP 2 Phage display (common panning) PMID:10725197 Anti-gG2 monoclonal antibody H7 Envelope glycoprotein G, gG, gG-2 Not determined. Not determined. f3-15mer phage display library (X15) 828 TPESTPLLPPFPRSV ALSSQGGMSPEPTPL DYTPQTSLELPPESF TPL 3 Phage display (common panning) PMID:10725197 Anti-gG2 monoclonal antibody F11 Envelope glycoprotein G, gG, gG-2 Not determined. Not determined. f3-15mer phage display library (X15) 829 YMQPDPPPPLHAPDY SPLPEPPPEHRALVP TSTPTEHTYPLEIIT P(3)-E-H 3 Phage display (common panning) PMID:10725197 Anti-gG2 monoclonal antibody E5 Envelope glycoprotein G, gG, gG-2 Not determined. Not determined. f3-15mer phage display library (X15) 830 GEASGLCCRWSSLKGC(30)[NA] GHREAFLPPWVFXFL(11)[NA] GEASGLCCRWSLRVV(8)[NA] YFTIPATLLPFGV(1)[NA] GNRPVRPQVYFSASS(1)[82%] SGFVAGKGFRLDFGS(1)[88%] 6 Phage display (common panning) 3 PMID:10649627 Anti-MAPS monoclonal antibody 9C10 Meningococcal group A polysaccharide (MAPS) Not determined. Not determined. f3-15mer phage display library (X15) ELISA Six phage clones were screened for MAPS mimicry by their ability to inhibit binding of human hyperimmune sera to nominal antigen, MAPS, by inhibition ELISA. The results of the inhibition ELISA with phage clone six (SGFVAGKGFRLDFGS) indicate that 5.5e10 TU of phage clone 6 inhibited the binding of human hyperimmune sera to MAPS (88% inhibition). The control phage, containing no insert, failed to inhibit binding in this assay. Phage clone 4 (GNRPVRPQVYFSASS) was also capable of inhibiting the binding of human hyperimmune sera to MAPS (82% inhibition). Other phage clone failed to inhibit binding of human hyperimmune sera to MAPS (data not shown). 831 RVVMSYPRHYWFSVR TRLGRIYWAAPSGIV ASRHAIRFIVFPAT LPLVFLTCLIMLSRV VSAAPTPAYWFGFYY 5 Phage display (common panning) PMID:10552746 Pancreatic alpha-amylase, PA Not determined. Not determined. Not determined. f3-15mer phage display library (X15) The two phage display ligands (RVVMSYPRHYWFSVR and TRLGRIYWAAPSGIV) with high-binding activities contained a high content of Arg, Tyr, and Trp residues with the short consensus sequence Arg-X-Tyr-Trp. These clones were shown to exhibit comparable binding interactions toward barley R-amylase based on transducing units titering and measurement of the dissociation constants. 832 ALSMWQPT LRWSQPTN APLTSPMM YNGRRKQV LLVRGLAP VSTVLGST KRVKSTSF HALAKRVV SRLMASSH YLVPNNN YEPVPVRG 11 Phage display (common panning) 5 PMID:11717916 Anti-c-myc protein monoclonal antibody 9E10 Myc proto-oncogene protein Not determined. Not determined. X8 phage display library The binding of peptides to 9E10 could be specifically inhibited by free c-myc decapeptide. 833 ASNHTHSSSIQF(3) EPIHRSTLTALL(1) EPIHRSTLTAPL(1) YQDMIYMRPLDS(1) DARHSSSLQMLF(1) FSLRPTMNFTNL(1) DPGKIYFHIAVS(1) PRPSPKMGVSV(1) TATHRHSSSI(1) 9 Phage display (common panning) 4 PMID:11694293 pFc' fragments of HAT Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 834 SHLGFDD(3) RQLVQPL(1) SPAPSDS(1) SSQSDPA(1) SKPTQLH(1) GTATSPH(1) TSDTGWR(1) 7 Phage display (common panning) 4 PMID:11694293 pFc' fragments of HAT Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The constant region of humanized anti-Tac (HAT), prepared by pepsin digestion and receptor-affinity chromatography, was used as the target for phage display in this study. 835 KPGDPSALHVVRCWIC(2) DGRRDVVVRSATFYL(1) EGLYSPWWPRSLPVL(1) SSKTRSFGVHLVGPY(1) LPPGLHVFPLASNRS(1) 5 Phage display (common panning) 3 PMID:11468163 Anti-GPIb monoclonal antibody 6B4 Platelet glycoprotein Ib alpha chain Not determined. Not determined. X15 phage display library 836 KPGEMR(3) CDTLKPGEC(3) CNKPGERTC(2) CADQKPGEC(2) CYKPGEWAC(1) CKPGEVQQC(1) CKPVENRAC(1) KPGEMRGAA(1) 8 Phage display (common panning) 3 PMID:11468163 Anti-GPIb monoclonal antibody 6B4 Platelet glycoprotein Ib alpha chain Not determined. Not determined. CX7C fUSE5 phage display library 837 PVLLFCFLAGRCVSV(7) LRCYSHCTFYFASST(1) 2 Phage display (common panning) 3 PMID:11468163 Anti-GPIb monoclonal antibody 12G1 Platelet glycoprotein Ib alpha chain Not determined. Not determined. X15 phage display library 838 MCPEMSFVAVHSCAR(15) 1 Phage display (common panning) 3 PMID:11468163 Anti-GPIb monoclonal antibody 12E4 Platelet glycoprotein Ib alpha chain Not determined. Not determined. X15 phage display library 839 GFLSSCNFRFCELVR(24) 1 Phage display (common panning) 3 PMID:11468163 Anti-GPIb monoclonal antibody 27A10 Platelet glycoprotein Ib alpha chain Not determined. Not determined. X15 phage display library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hage display (common panning) 4 PMID:10064083 HLA class II histocompatibility antigen, DRB1-1 beta chain HLA class II histocompatibility antigen, DM Not determined. Not determined. M13lib phage display library ELISA Purified DR1 molecules (35 nM) were incubated with the N-terminal biotinylated peptide GFKA7 (0.2 μM) and competitor peptides in concentrations ranging from 0.001 to 100 μM. Competitive peptide binding to DR1 was analyzed using the high-throughput ELISA-based assay. IC50 (μM) values represent the concentration of peptide competitors yielding 50 % binding inhibition of biotinylated GFKA7 to DR1. 841 GPRPPSP(1) GPRPTGH(1) GPRPHTP(1) GPRPPAP(1) GPRPPSH(1) GPRPPMS(1) GPRPHLM(1) GPRPNLT(1) GPRPPYA(1) GPRPATM(1) GPRPVNP(1) LHSTTFW(1) KHATTFW(1) EHSLTFW(1) DHVNTFW(1) ATPIRQP(1) ASTSESL(1) GPRP 17 Phage display (common panning) 1 PMID:10677284 High molecular mass serum fraction Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Here this procedure is used to probe a mixture of high molecular mass components of human cord blood with phage-peptide display libraries. The initial panning recovered phage that bore the consensus motif Gly-Pro-Arg-Pro, a known fibrinogen-binding motif. These phage bound specifically to purified fibrinogen. A series of peptides containing the Gly-Pro-Arg-Pro motif efficiently blocked the binding of phage having the same motif, presumably by binding to their common target. 842 HLHGTPRPSSGL(12) HLHGTPRMLPPL(3) GPRPPLPPALPL(2) GPRPPVAGSWPF(2) GPRPWPPQELSR(1) GPRPPAALPHPL(1) GPRPPSLPPDPA(1) GPRPSMGLATNL(1) GPRPLNPPDKLP(1) GPRPVYPRHDYT(1) GPRPPSATAFPP(1) GPRPAHPPPLTH(1) GPRPPASNFSFG(1) GPRPHTILTSPS(1) GPRPATTWYPNS(1) GPRPSLSHAHNW(1) GPRPAWTSPTFI(1) GPRPPSTHWMQQ(1) GPRPNFPSLPTQ(1) GPRPNPLTVALH(1) GPRPPITMPMFM(1) GPRPVHNTFYYA(1) GPRPATYMPPSL(1) GPRPLYTLSPHT(1) LLAGPRPPSMHV(1) GPRP 25 Phage display (common panning) 1 PMID:10677284 High molecular mass serum fraction Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Here this procedure is used to probe a mixture of high molecular mass components of human cord blood with phage-peptide display libraries. The initial panning recovered phage that bore the consensus motif Gly-Pro-Arg-Pro, a known fibrinogen-binding motif. These phage bound specifically to purified fibrinogen. A series of peptides containing the Gly-Pro-Arg-Pro motif efficiently blocked the binding of phage having the same motif, presumably by binding to their common target. 843 CAGPRPNPS(13) CAGPRPALG(6) CAGPRPPHC(1) CKTNDNTQC(1) CQGDPSHHC(1) CLNTSSMTC(1) CSTAVGPQC(1) CELSARGMC(1) CNHQHNNAC(1) CKSGLTPTC(1) CRVNFQNVC(1) GPRP 11 Phage display (common panning) 1 PMID:10677284 High molecular mass serum fraction Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Here this procedure is used to probe a mixture of high molecular mass components of human cord blood with phage-peptide display libraries. The initial panning recovered phage that bore the consensus motif Gly-Pro-Arg-Pro, a known fibrinogen-binding motif. These phage bound specifically to purified fibrinogen. A series of peptides containing the Gly-Pro-Arg-Pro motif efficiently blocked the binding of phage having the same motif, presumably by binding to their common target. 844 GHWKWYE(1) SHYMRMQ(1) NHYMSWQ(1) NHYTPLW(1) THMIEFW(1) SHFQNFW(1) SHYRNPQ(1) SHYTQFY(1) HHYTLPQ(1) SHYPYWE(1) HHADGDT(1) NHHAMFW(1) SHY 12 Phage display (common panning) 2 PMID:10677284 High molecular mass serum fraction Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Here this procedure is used to probe a mixture of high molecular mass components of human cord blood with phage-peptide display libraries. The initial panning recovered phage that bore the consensus motif Gly-Pro-Arg-Pro, a known fibrinogen-binding motif. These phage bound specifically to purified fibrinogen. A second round of panning was performed against the same target mixture in the presence of this blocking peptide. Phage recovered from this second panning exhibited a motif (Ser-His-Tyr) that was subsequently shown to bind specifically to complement component C1q. A second peptide containing this motif specifically blocked the interaction of the phage with C1q. 845 FHWSWYTPSRPS(2) NHMSRWEAWDRL(2) HLHGTPRMLPPL(2) HLHGTPRPSSGL(2) AHLRQDSAWKLN(1) SHTPDRPGAFYA(1) SHYTSTWAASEG(1) SHYPQELWAGSS(1) SHY 8 Phage display (common panning) 2 PMID:10677284 High molecular mass serum fraction Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Here this procedure is used to probe a mixture of high molecular mass components of human cord blood with phage-peptide display libraries. The initial panning recovered phage that bore the consensus motif Gly-Pro-Arg-Pro, a known fibrinogen-binding motif. These phage bound specifically to purified fibrinogen. A second round of panning was performed against the same target mixture in the presence of this blocking peptide. Phage recovered from this second panning exhibited a motif (Ser-His-Tyr) that was subsequently shown to bind specifically to complement component C1q. A second peptide containing this motif specifically blocked the interaction of the phage with C1q. 846 CAHHLKQFW(1) CAAHYGYEW(1) CWFAPHLSC(1) CWFAPHLRC(1) 4 Phage display (common panning) 2 PMID:10677284 High molecular mass serum fraction Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Here this procedure is used to probe a mixture of high molecular mass components of human cord blood with phage-peptide display libraries. The initial panning recovered phage that bore the consensus motif Gly-Pro-Arg-Pro, a known fibrinogen-binding motif. These phage bound specifically to purified fibrinogen. A second round of panning was performed against the same target mixture in the presence of this blocking peptide. Phage recovered from this second panning exhibited a motif (Ser-His-Tyr) that was subsequently shown to bind specifically to complement component C1q. A second peptide containing this motif specifically blocked the interaction of the phage with C1q. 847 FNPFLLD(9) YWNPFFL(4) FNPFSAG(1) SYALRAP(1) ALGLSPL(1) KVWIPQK(1) NPL 6 Phage display (common panning) 3 PMID:10677284 High molecular mass serum fraction Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Here this procedure is used to probe a mixture of high molecular mass components of human cord blood with phage-peptide display libraries. The initial panning recovered phage that bore the consensus motif Gly-Pro-Arg-Pro, a known fibrinogen-binding motif. These phage bound specifically to purified fibrinogen. A second round of panning was performed against the same target mixture in the presence of this blocking peptide. Phage recovered from this second panning exhibited a motif (Ser-His-Tyr) that was subsequently shown to bind specifically to complement component C1q. A third round of panning performed in the presence of both the fibrinogen- and the C1q- blocking peptides yielded phage with a new peptide motif (Asn-Pro-Phe) that also bound specifically to C1q, apparently at a new site. 848 CRYRGDLGRRC(8) 1 Phage display (common panning) 4 PMID:11006114 Madin-Darby canine kidney cells strain II, MDCKII Not determined. Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) MDCK strain II (MDCKII) cells transfected with a rabbit receptor for polymeric immunoglobulins (pIgR) were kindly provided by Dr. K. E. Mostov (University of California). The identified peptide contained a putative integrinbinding (RGD) motif suggesting the involvement of integrins, but not pIgR, in phage transcytosis. The paper data suggest the feasibility of using short peptides for targeting transcytotic pathways nand facilitating delivery of macromolecules across cellular barriers. 849 SVSVGMKPSPRP(16) 1 Phage display (competitive panning) 5 PMID:10856231 Tobacco plasmodesmal-enriched cell wall fraction, W2 cell fraction Homeotic protein knotted-1 Not determined. Not determined. Ph.D.-12 phage display library (X12) Elution of bound phages was achieved by addition of KN1. A KN1 peptide antagonist had the capacity to interact with a motif involved in the dilation of plasmodesmal microchannels. 850 QDVHLTQQSRYT(7) SFHLELLERGQS(6) LSTHTTESRSMV(1) DPLNKMLLSQDL(1) 4 Phage display (competitive panning) 5 PMID:10856231 Tobacco plasmodesmal-enriched cell wall fraction, W2 cell fraction Movement protein, MP Not determined. Not determined. Ph.D.-12 phage display library (X12) Elution of bound phages was achieved by addition of CMV-MP. 851 RFDSLKV(6) GRGDGDV(2) 2 Phage display (subtractive panning) 4 PMID:10913838 Human foetal tracheal epithelial cell line CFT-2 Not determined. Not determined. Not determined. HyC, HyB and HyA phage display library pool Before incubation with the cells, the phage libraries were subtracted to remove phage particles that absorb to polystyrene and to serum proteins. 852 FRTAISGTPQFY(6) SYRTPITGTLIT(2) HSHTHKALAGTP(1) LVAKPHMRTPNL(1) WHWQYTPWWRGS(1) 5 Phage display (common panning) 4 PMID:10880841 Rabbit hyperimmune serum against peptide (MSESHVKISRTIIRGTSPST) AA1–20 of Taenia solium paramyosin (MSESHVKISRTIIRGTSPST) Not determined. Not determined. Ph.D.-12 phage display library (X12) Rabbit hyperimmune serum, diluted 1:50 in PBS to give a final volume 300 ml, was distributed in 6 wells (50 ml:well) of 96-well flat bottom polystyrene microtitration plates. Eight clones presented two sequences with internal RT-I-GT motif and three peptides contained only a part of this motif: RT or GT/S, or T-RGS. Comparison with the amino-terminal 20aa sequence of the TPmy highlighted the Arg10-Thr16 region of apparent homology to the motif shared by two of these peptides and partially by three of them. 853 SLSRMPIIGTSA(4) AWTHTLIRLPD(2) NEMTLIRMNMAA(2) DIRQPIIGTLHP(1) HIARTPIAGTNL(1) HRVPISGTSA(1) LEPTLIRLPQTL(1) MKETHSTLHQP(1) TLIR 8 Phage display (common panning) 3 PMID:10880841 Anti-peptide (MSESHVKISRTIIRGTSPST) polyclonal antibody IgG AA1–20 of Taenia solium paramyosin (MSESHVKISRTIIRGTSPST) Not determined. Not determined. Ph.D.-12 phage display library (X12) Peptide sequences was selected on panning of the 12-mer library in solution. A total of 300 ng of purified IgG fraction were mixed with the library (2e10 phage virions) and incubated 20min at room temperature. Then 50 ml of protein G-Agarose suspension (Gibco BRL) were added and the incubation was continued 15 min at the same temperature. The incubation mixture was centrifuged, supernatant discarded and the pellet washed 10 times with PBS-T. The final pellet was resuspended in 1 ml of glycine-HCl, pH 2.2 and after 10 min incubation at room temperature the eluate was collected after centrifugation The phage were titered, amplified and the selection was repeated two more times, first with 100 ng and then with 30 ng of IgG except for the third round where phage were eluted in two steps, first at pH 4.0 and then at pH 2.2. 854 FPNRTPISGTTW(1) 1 Phage display (common panning) 4 PMID:10880841 Anti-peptide (MSESHVKISRTIIRGTSPST) polyclonal antibody IgG AA1–20 of Taenia solium paramyosin (MSESHVKISRTIIRGTSPST) Not determined. Not determined. Ph.D.-12 phage display library (X12) Peptide sequences was selected on panning of the 12-mer library in solution. A total of 300 ng of purified IgG fraction were mixed with the library (2e10 phage virions) and incubated 20min at room temperature. Then 50 ml of protein G-Agarose suspension (Gibco BRL) were added and the incubation was continued 15 min at the same temperature. The incubation mixture was centrifuged, supernatant discarded and the pellet washed 10 times with PBS-T. The final pellet was resuspended in 1 ml of glycine–HCl, pH 2.2 and after 10 min incubation at room temperature the eluate was collected after centrifugation The phage were titered, amplified and the selection was repeated two more times, first with 100 ng and then with 30 ng of IgG except for the third round where phage were eluted in two steps, first at pH 4.0 and then at pH 2.2. 855 KAAQIYSEP(8)[0.684, 20.0 ± 6.0] DYDGYGWRE(3)[0.360, 9.3 ± 0.2] YAADITNGL(3)[0.648, 11.0 ± 2.3] NKPNWDGYA(1)[0.648, 10.1 ± 0.2] IQWDGYARQ(1)[0.432, ND] WSSGYGTGR(1)[0.234, 8.2 ± 1.0] AYESGYNLP(1)[0.252, 9.7 ± 1.5] CSAGICFSE(1)[0.414, 7.2 ± 1.6] [DS]-G-Y-[AG], A(2)-A-x-I 8 Phage display (common panning) 2 PMID:10759860 Anti-CA monoclonal antibody A-56.36 Crotoxin acid chain, CA Not determined. Not determined. X9 and CX9C phage display library pool ELISA ELISA measurements of the binding to A-56.36 expressed as A490, SD ± 10%. ELISA measurements of the competition with CA, expressed as the inhibition percentage of the binding of A-56.36 to CA-coated plates in the presence of a fixed amount of phages (e12 p.f.u./mL). Average values and SD on two experiments are shown. ND denotes not determined. By screening random phage-displayed libraries with the mAb, phagotopes bearing the (D/S)GY(A/G) or AAXI consensus motifs were selected. They all bound A-56.36 in ELISA and competed with CA for mAb binding, although with different reactivities. 856 DFFEII(1) DFLMLV(1) QFVFCW(1) DFLVQL(1) DFLAIV(1) 5 Phage display (common panning) 2 PMID:1696028 Anti-Myohemerythrin monoclonal antibody A2 Myohemerythrin, MHr Not determined. Not determined. f3-6mer phage display library (X6) In the first round, 1e12 phage particles were incubated overnight with 1000nM bio-MAb in 10 microliter reaction mixtures; the phages were diluted and reacted 10 to 20 minutes on streptavidin-coated dishes; the dishes were extensively washed; and the bound phages were eluted in acid buffer. Phages eluted in the first round were amplified on agar medium and subjected to second round of paning with the same bio-MAb at concentrations of 10 nM. 857 DFLAIV(4) DFLEYI(2) DFLEIL(1) DFLEIV(1) 4 Phage display (common panning) 2 PMID:1696028 Anti-Myohemerythrin monoclonal antibody A2 Myohemerythrin, MHr Not determined. Not determined. f3-6mer phage display library (X6) In the first round, 1e12 phage particles were incubated overnight with 1000nM bio-MAb in 10 microliter reaction mixtures; the phages were diluted and reacted 10 to 20 minutes on streptavidin-coated dishes; the dishes were extensively washed; and the bound phages were eluted in acid buffer. Phages eluted in the first round were amplified on agar medium and subjected to second round of paning with the same bio-MAb at concentrations of 0.1 nM. 858 DFLAWL(14) DFLEML(2) 2 Phage display (common panning) 3 PMID:1696028 Anti-Myohemerythrin monoclonal antibody A2 Myohemerythrin, MHr Not determined. Not determined. f3-6mer phage display library (X6) In the first round, 1e12 phage particles were incubated overnight with 1000nM bio-MAb in 10 microliter reaction mixtures; the phages were diluted and reacted 10 to 20 minutes on streptavidin-coated dishes; the dishes were extensively washed; and the bound phages were eluted in acid buffer. Phages eluted in the first round were amplified on agar medium and subjected to second round of paning with the same bio-MAb at concentrations of 0.1 nM. Phages from the second round were amplified and subjected to third round with 0.1 nM bio-MAb. 859 DFLEWL(4) DFMEWL(1) DFLEQL(1) DFLHFI(1) AWERRG(1) 5 Phage display (common panning) 2 PMID:1696028 Anti-Myohemerythrin monoclonal antibody M33 Myohemerythrin, MHr Not determined. Not determined. f3-6mer phage display library (X6) In the first round, 1e12 phage particles were incubated overnight with 1000nM bio-MAb in 10 microliter reaction mixtures; the phages were diluted and reacted 10 to 20 minutes on streptavidin-coated dishes; the dishes were extensively washed; and the bound phages were eluted in acid buffer. Phages eluted in the first round were amplified on agar medium and subjected to second round of paning with the same bio-MAb at concentrations of 10 nM. 860 CRFVWC(2) DFLERI(1) CELEKC(1) CRLEKC(1) DFMEWL(1) DFLVQL(1) 6 Phage display (common panning) 2 PMID:1696028 Anti-Myohemerythrin monoclonal antibody M33 Myohemerythrin, MHr Not determined. Not determined. f3-6mer phage display library (X6) In the first round, 1e12 phage particles were incubated overnight with 1000nM bio-MAb in 10 microliter reaction mixtures; the phages were diluted and reacted 10 to 20 minutes on streptavidin-coated dishes; the dishes were extensively washed; and the bound phages were eluted in acid buffer. Phages eluted in the first round were amplified on agar medium and subjected to second round of paning with the same bio-MAb at concentrations of 0.1 nM. 861 DFLEWL(16) DFLEML(1) 2 Phage display (common panning) 3 PMID:1696028 Anti-Myohemerythrin monoclonal antibody M33 Myohemerythrin, MHr Not determined. Not determined. f3-6mer phage display library (X6) In the first round, 1e12 phage particles were incubated overnight with 1000nM bio-MAb in 10 microliter reaction mixtures; the phages were diluted and reacted 10 to 20 minutes on streptavidin-coated dishes; the dishes were extensively washed; and the bound phages were eluted in acid buffer. Phages eluted in the first round were amplified on agar medium and subjected to second round of paning with the same bio-MAb at concentrations of 0.1 nM. Phages from the second round were amplified and subjected to third round with 0.1 nM bio-MAb. 862 RWWWPTR RWWWLPR RWWWFPR KWWMASR RWWWDPF RWWWSFS VKWWWSA IWKWWWK 8 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab003 Not determined. Not determined. Not determined. X7 phage display library The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 863 FCSFCVL FCSDCIL FCGDCVL 3 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab004 Not determined. Not determined. Not determined. X7 phage display library The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 864 CRWWSGQPC CRWWWQDTC CRWWMGVSC CRWWNGSWC CRWWMSYTC CRWWRGQGC CRWWFGQFC CRWWRGEVC CRWWLGSAC CRWWAGSTC CRWWWVETC CKWWGGRGC 12 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab003 Not determined. Not determined. Not determined. CX7C phage display library The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 865 WNWPLPPVRQFS WPWPLPPEPPLG SWYWPLPPWRLG SWPWYHPHIKSH 4 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab001 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 866 TPEKWHRLLTMS ATPWSQWLDAPR 2 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab003 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 867 EMSVDWWSPISS ASSVDWWPVRPP FMWPDQNPRHSM 3 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab004 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 868 QLLFPSSPRAPAPWTFTF 1 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab006 Not determined. Not determined. Not determined. X18 phage display library The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 869 YYCYFTEAPYSYWGNLVC SQSPPRYWAWCAGYWCEL 2 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab007 Not determined. Not determined. Not determined. X18 phage display library The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 870 SYWPCHPGTRHCSNRV 1 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab003 Not determined. Not determined. Not determined. Beta-sheet (BS) phage display library (X4CX6CX4) The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 871 PVLVCGPKWSNCSPAN SPSLCWPWLEQCTEGI GDQPCPIFDRECHKPT GIGPCELIDSECEASE 4 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab004 Not determined. Not determined. Not determined. Beta-sheet (BS) phage display library (X4CX6CX4) The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 872 YHRWCVMDRRACFEAP NLSECAMPTRKCSRTA 2 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab005 Not determined. Not determined. Not determined. Beta-sheet (BS) phage display library (X4CX6CX4) The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 873 LEYACNGPSQLCSYVR LPWACPFTGWFCDLIT TEKTCAGPGDLCLLVR AQRKCAGNWAICRLVY 4 Phage display (subtractive panning) 3/4 PMID:21117166 Chronic Lymphocytic Leukemia Fragment Antigen-Binding (Fab) Fragments Fab006 Not determined. Not determined. Not determined. Beta-sheet (BS) phage display library (X4CX6CX4) The library was screened on BCR Fab fragments after a 2-fold negative selection on polyclonal human immunoglobulins (Octapharma, Lachen, Switzerland). 874 SWLSRRPLPPLPP(2) VPLARRPLPPLPP(2) SSLRFRPLPPLPP(2) WSLFHRPLPPLPP(1) CLLCSRPLPPLPP(1) IHLRTRPLPPLPP(1) PHPARRPLPPLPP(1) VSLAQRPLPPLPP(1) ISLARRPLPPLPP(1) SWLANRPLPPLPP(1) LSLQMRPLPPLPP(1) SSLSRRPLPPLPP(1) LSLAQRPLPPLPP(1) LSLFRRPLPPLPP(1) SWLSHRPLPPLPP(1) SSLCGRPLPPLPP(1) 16 Phage display (common panning) 4 PMID:7479908 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. N+5 class I phage display library GST-Src SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 875 SAPSRRPLPPLPPP(1) LRWSARPLPPLPPP(1) KALASRPLPPLPPP(1) TTLSARPLPPLPPP(1) SYLSRRPLPPLPPP(1) LSPMSRPLPPLPPP(1) LPMARRPLPPLPPP(1) ASFASRPLPPLPPP(1) VGLSNRPLPPLPPP(1) IPIGHRPLPPLPPP(1) IALSTRPLPPLPPP(1) FFLASRPLPPLPPP(1) LDRSRRPLPPLPPP(1) RWITSRPLPPLPPP(1) GLGARRPLPPLPPP(1) DDLSGRPLPPLPPP(1) NPLLRRPLPPLPPP(1) 17 Phage display (common panning) 4 PMID:7479908 Fyn SH3 domain of tyrosine-protein kinase Fyn Not determined. Not determined. Not determined. N+5 class I phage display library GST-Fyn SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 876 SWLIARPLPPLPPP(1)[NT] RALPPRPLPPLPPP(1)[NT] WSIRSRPLPPLPPP(1)[NT] YPFHGRPLPPLPPP(1)[NT] PWLLRRPLPPLPPP(1)[NT] SWLSVRPLPPLPPP(1)[NT] SWLGSRPLPPLPPP(1)[NT] PWWLDRPLPPLPPP(1)[1451.141] LTMAGRPLPPLPPP(1)[NT] DFMASRPLPPLPPP(1)[NT] DWYHSRPLPPLPPP(1)[NT] LSFSNRPLPPLPPP(1)[NT] MDLLRRPLPPLPPP(1)[NT] AAMVGRPLPPLPPP(1)[NT] RLLPPRPLPPLPPP(1)[NT] SWVACRPLPPLPPP(1)[NT] PWPVGRPLPPLPPP(1)[NT] 17 Phage display (common panning) 4 PMID:7479908 Lyn SH3 domain of tyrosine-protein kinase Lyn Not determined. Not determined. Not determined. N+5 class I phage display library Surface plasmon resonance (SPR) Binding of phage-displayed peptides to Src was measured by BIAcore technology. The amount of phage binding to immobilized SH3 domain is given as arbitrary resonance units (RU) and has been corrected for bulk refractive-index contributions of the unbound phage. Data shown were reproduced from the graph. NT denotes not tested. GST-Lyn SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 877 SALARRPLPPLPPP(2) PNLRTRPLPPLPPP(1) LPLSRRPLPPLPPP(1) SWWSRRPLPPLPPP(1) TFLFRRPLPPLPPP(1) THPARRPLPPLPPP(1) FPVAQRPLPPLPPP(1) PSPRSRPLPPLPPP(1) RSLSMRPLPPLPPP(1) VLLALRPLPPLPPP(1) SVLARRPLPPLPPP(1) NRAAKRPLPPLPPP(1) SSLAQRPLPPLPPP(1) TPLGRRPLPPLPPP(1) PNPSYRPLPPLPPP(1) SASARRPLPPLPPP(1) SWMARRPLPPLPPP(1) SPLARRPLPPLPPP(1) 18 Phage display (common panning) 4 PMID:7479908 Tyrosine-protein kinase Yes Not determined. Not determined. Not determined. N+5 class I phage display library GST-Yes SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 878 CLLTTRPLPPLPPP(7)[NT] CLNCFRPLPPLPPP(6)[984.609] CLNCSRPLPPLPPP(5)[NT] LLSGPRPLPPLPPP(4)[NT] CLNCLRPLPPLPPP(4)[NT] FKRPPRPLPPLPPP(1)[NT] FKRPFRPLPPLPPP(1)[NT] LVLPRRPLPPLPPP(1)[NT] FHRPNRPLPPLPPP(1)[NT] RRVPPRPLPPLPPP(1)[NT] WSRTKRPLPPLPPP(1)[NT] RVRPSRPLPPLPPP(1)[NT] RYLPWRPLPPLPPP(1)[NT] CCLTTRPLPPLPPP(1)[NT] RPFPRRPLPPLPPP(1)[NT] RVPIRRPLPPLPPP(1)[NT] ARLPPRPLPPLPPP(1)[NT] RYLSWRPLPPLPPP(1)[NT] 18 Phage display (common panning) 4 PMID:7479908 PI3K SH3 domain of PI3-kinase subunit p85-alpha Not determined. Not determined. Not determined. N+5 class I phage display library Surface plasmon resonance (SPR) Binding of phage-displayed peptides to Src was measured by BIAcore technology. The amount of phage binding to immobilized SH3 domain is given as arbitrary resonance units (RU) and has been corrected for bulk refractive-index contributions of the unbound phage. Data shown were reproduced from the graph. NT denotes not tested. GST-PI3K SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 879 RSLRPLPPPPVPSL(12)[352.868] RSLRPLPLPPVPSL(1)[139.410] RSLPLPPLPARPHP(1)[NT] RSFRPLPPLPTLLPS(1)[NT] RSLRPLPPLPTPPLH(1)[NT] 5 Phage display (common panning) 4 PMID:7479908 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. C+5 class I phage display library Surface plasmon resonance (SPR) Binding of phage-displayed peptides to Src was measured by BIAcore technology. The amount of phage binding to immobilized SH3 domain is given as arbitrary resonance units (RU) and has been corrected for bulk refractive-index contributions of the unbound phage. Data shown were reproduced from the graph. NT denotes not tested. GST-Src SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 880 RSLRPLPPLPLPPLT(1) RSLRPLPPLPSAPRV(1) RSLRPLPPLPVLSEP(1) RSLRPLPPLPTSTQP(1) RSLRPLPPLPHLPDS(1) RSLRPLPPLPSYTSH(1) RSLRPLPPLPVATHP(1) RSLRPLPPLPSSLSR(1) RSLRPLPPLPAFHAR(1) RSFRPLPPLPTVASP(1) 10 Phage display (common panning) 4 PMID:7479908 Fyn SH3 domain of tyrosine-protein kinase Fyn Not determined. Not determined. Not determined. C+5 class I phage display library GST-Fyn SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 881 RSLRPLPPLPLPPRL(1)[NT] RSLRPLPPLPFPRYL(1)[NT] RSLRPLPPLPPSSAP(1)[NT] RSLRPLPPLPPRAPF(1)[NT] RSLRPLPPLPPRPPF(1)[377.387] RSLRPLPPLPSRPGP(1)[NT] RSLRPLPPLPPRPAY(1)[NT] 7 Phage display (common panning) 4 PMID:7479908 Lyn SH3 domain of tyrosine-protein kinase Lyn Not determined. Not determined. Not determined. C+5 class I phage display library Surface plasmon resonance (SPR) Binding of phage-displayed peptides to Src was measured by BIAcore technology. The amount of phage binding to immobilized SH3 domain is given as arbitrary resonance units (RU) and has been corrected for bulk refractive-index contributions of the unbound phage. Data shown were reproduced from the graph. NT denotes not tested. GST-Lyn SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 882 RSLRPLPPLPLPPRH(9)[477.953] RSLRPLPPLPLPPRT(1)[357.737] RSLRPLPPLPLPPRL(1)[NT] RSLRPLPPLPLPPPH(1)[NT] 4 Phage display (common panning) 4 PMID:7479908 PI3K SH3 domain of PI3-kinase subunit p85-alpha Not determined. Not determined. Not determined. C+5 class I phage display library Surface plasmon resonance (SPR) Binding of phage-displayed peptides to Src was measured by BIAcore technology. The amount of phage binding to immobilized SH3 domain is given as arbitrary resonance units (RU) and has been corrected for bulk refractive-index contributions of the unbound phage. Data shown were reproduced from the graph. NT denotes not tested. GST-PI3K SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 883 GAAPPLPPRNPPRT(2) GAAPPLPPRNKPRL(2) GAAPPLPPRNPPRL(1) GAAPPLPPRNKVRG(1) GAAPPLPPRNQTFR(1) GAAPPLPPRNRLHR(1) GAAPPLPPRNVPRL(1) GAAPPLPPRNRLHA(1) GAAPPLPPRNRVKL(1) GAAPPLPPRNRVRL(1) GAAPPLPPRNRVVL(1) GAAPPLPPRNIPRV(1) GAAPPLPPRNPPRK(1) GAAPPLPPRNKARL(1) 14 Phage display (common panning) 4 PMID:7479908 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. C+5 class II phage display library GST-Src SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 884 GAAPPLPPRNRVRL(2) GAAPPLPPRNPPRV(1) GAAPPLPPRNPRPI(1) GAAPPLPPRNAVRL(1) GAAPPLPPRNRPHF(1) GAAPPLPPRNPKHR(1) GAAPPLPPRNPPRF(1) GAAPPLPPRNSRPL(1) GAAPPLPPRNPARL(1) GAAPPLPPRNTPRL(1) GAAPPLPPRNPRPL(1) GAAPPLPPRNPPRT(1) GAAPPLPPRNPPTV(1) 13 Phage display (common panning) 4 PMID:7479908 Fyn SH3 domain of tyrosine-protein kinase Fyn Not determined. Not determined. Not determined. C+5 class II phage display library GST-Fyn SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 885 GAAPPLPPRTYLLP(2) GAAPPLPPRTYGTT(1) GAAPPLPPRPPWMM(1) GAAPPLPPRTRYSL(1) GAAPPLPPRPPWMT(1) GAAPPLPPRTYWMP(1) GAAPPLPPRPTFMA(1) GAAPPLPPRPTWMT(1) GAAPPLPPRTYLMP(1) GAAPPLPPRPAWMA(1) GAAPPLPPRSYLLP(1) GAAPPLPPRTTPYT(1) GAAPPLPPRPSWMS(1) GAAPPLPPRPVTWI(1) GAAPPLPPRPSFMS(1) GAAPPLPPRPAFFS(1) GAAPPLPPRPSFLQ(1) 17 Phage display (common panning) 4 PMID:7479908 Lyn SH3 domain of tyrosine-protein kinase Lyn Not determined. Not determined. Not determined. C+5 class II phage display library GST-Lyn SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 886 GAAPPLPPRNPPRT(2) GAAPPLPPRNPPRI(2) GAAPPLPPRNRPRV(1) GAAPPLPPRNPKHR(1) GAAPPLPPRNPPRS(1) GAAPPLPPRNPKHW(1) GAAPPLPPRNPVRV(1) GAAPPLPPRNKARL(1) GAAPPLPPRNKHPP(1) GAAPPLPPRNPPRK(1) GAAPPLPPRNKRHL(1) GAAPPLPPRKHLSS(1) GAAPPLPPRNPPRV(1) GAAPPLPPRNKLRV(1) 14 Phage display (common panning) 4 PMID:7479908 Tyrosine-protein kinase Yes Not determined. Not determined. Not determined. C+5 class II phage display library GST-Yes SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 887 GAAPPLPPRPPRPA(2) GAAPPLPPRPPRPL(2) GAAPPLPPRPPLRS(1) GAAPPLPPRPPIRK(1) GAAPPLPPRPPLPF(1) GAAPPLPPRPPKPA(1) GAAPPLPPRPPRPP(1) GAAPPLPPRPPLRQ(1) GAAPPLPPRRALRL(1) GAAPPLPPRRLRVG(1) GAAPPLPPRPPRKP(1) GAAPPLPPRPPRTP(1) GAAPPLPPRPPRPS(1) GAAPPLPPRPPRPF(1) 14 Phage display (common panning) 4 PMID:7479908 PI3K SH3 domain of PI3-kinase subunit p85-alpha Not determined. Not determined. Not determined. C+5 class II phage display library GST-PI3K SH3 was produced in Escherichia coli strain BL21, purified and immobilized on polystyrene as the target. 888 CHPQGPPC(56)[8.1, 0.230] CHPQFPLC(4)[NT] CHPQFTLC(2)[NT] CHPQFSNC(1)[14.0, 0.400] CHPQFNLC(1)[NT] CHPQGDRC(1)[NT] CHPQFRHC(1)[NT] CHPQSGRC(1)[NT] HPQ 8 Phage display (common panning) 3 PMID:7492543 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, 1VWF,1VWG,1VWH,1VWI,1VWJ,1VWK,1VWL,1VWO,1VWP,1VWQ,1VWR,1SLE, CX6C M13 phage display library Surface plasmon resonance (SPR) KD (μM) and IC50 ((μM)) values were shown. Streptavidin-coated paramagnetic particles (Promega, Madison, WI) were used in the biopanning procedure. HPQGPPC showed the highest affinity, which was 3 orders of magnitude higher than those of the linear peptides such as FSHPQNT and HDHPQNL, identified previously. The structure of HPQGPPC-Streptavidin complex have been solved (1SLE). 889 CHPQFPC(28)[32.6, 0.930] CHPQFNC(4)[246.0, 7.0] CHPQVPC(4)[NT] CHPQVAC(3)[NT] CHPQFMC(2)[NT] CHPQFRC(2)[NT] CHPQNAC(1)[NT] CHPQVSC(1)[NT] CHPQFAC(1)[NT] CHPQVRC(1)[NT] CWHPQFC(1)[NT] HPQ 11 Phage display (common panning) 3 PMID:7492543 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. CX5C M13 phage display library Surface plasmon resonance (SPR) KD (μM) and IC50 ((μM)) values were shown. Streptavidin-coated paramagnetic particles (Promega, Madison, WI) were used in the biopanning procedure. 890 CHPQFC(20)[16.0, 0.47] HPQ 1 Phage display (common panning) 3 PMID:7492543 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, 1SLD,1VWB,1VWC,1VWD,1VWE,1VWM,1VWN, CX4C M13 phage display library Surface plasmon resonance (SPR) KD (μM) and IC50 ((μM)) values were shown. Streptavidin-coated paramagnetic particles (Promega, Madison, WI) were used in the biopanning procedure. The structure of CHPQFC-Streptavidin complex have been solved (1SLD). 891 ISFENTWLWHPQFSS(5) LNHPMDNRLHVSTSP(4) SDDWWHDHPQNLRSS(3) PCHPQYRLCQRPLKQ(2) QPFLHPQGDERWYMI(2) MLWYSPHSFSHPQNT(1) SWWLSWHPQNTKELG(1) LCHPQFPRCNLFRKV(1) ALCCLSSPHPNGAIF(1) HPQ 9 Phage display (common panning) 2 PMID:2143033 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. X15 M13 phage display library Streptavidin-coated polystyrene plates were used as an affinity matrix for panning. 892 GAVITH(1) RDIVVA(1) VYSHAS(1) GSYSAG(1) LDIVVA(1) 5 Phage display (subtractive panning) 7 PMID:7496483 Chymotrypsinogen A Not determined. Not determined. Not determined. f3-6mer phage display library (X6) Since BSA was used as a blocking agent in the immobilization procedure, an addition of BSA was made in the seventh round to exclude possible BSA-binding phage clones. 893 VIRYKDVRKYYLGPARAVLT(3) FQKETLMRYWGPGRYFTAY(2) MVNGRSRAVFYYLGPGRLKM(2) SNAVLSGHPQNDYQAFVALR(2) VYAVKQGIRLRYYVGPGRIL(1) EIYGAQGAVGVLFGPGRVWL(1) AAPAGVLHIRHLVGPNRYNF(1) GIFLSEGHSPVNVGPNRLKV(1) VLGAQNTKHLGPGRFWYLL(1) SLVNIPRYFVGPGRRPMLVL(1) SLVPSPLLYIGPGRLRMTDH(1) SVELRSIQLIGIGRNFHWMG(1) MHSFYKGPGRRGYDTLWNQF(1) YILGPGRGRFPGVNCMPYI(1) NTFALVPDWLGSPHGHSHHP(1) PTDTWQAFWLRHPQNHVFLA(1) [YL]-[VLI]-G-P-G-R-x-F 16 Phage display (common panning) 2 PMID:7506691 Anti-HIV-l isotype MN gp120 monoclonal antibody 58.2 Surface protein gp120 of Envelope glycoprotein gp160 Not determined. Not determined. PDL-20 phage display library SA-bio biopanning protocol: mixing phage with biotinylated anti-HIV-l mAB 58.2 in solution overnight at 4°C and then binding to SA coated plate. The bound bacteriophage were washed 12 times with 0.5% Tween20 in PBS, eluted by treatment with 0.1 M HCl, adjusted to pH 2.2 with glycine and neutralized by addition of 1M Tris base. The phage were enumerated by titering transducing units and the phage population amplified by growth in yeast tryptone broth containing 20 p/ml of tetracycline. In subsequent cycles, the biotinylated mAb 58.2 was added to the SA coated plates for 2h at 4°C and the free SA blocked by the addition of bio for 1h at 4°C before the addition of the phage. Washing, elution and amplification of bacteriophage were as above. 894 MDHLGPGRYHWFNRNDRVFK(4) GVRHMGPGRASFDEFQSLHV(3) WVFSPSTGGHRLYTGPGRHF(2) RFMSAYVMKIGPNRHGFRAP(2) RSQGLMVLGPGRGFMIFNAG(2) RRWCRSPNNLFIASPHGPLR(1) AAPAGVLHIRHLVGPNRYNF(1) NLRRTSIRAFYMLGPSRWMP(1) VRKPFQFAASLGPHRAFSHW(1) RAKWNKHLYFEGPSRMFNFT(1) SLVPSPLLYIGPGRLRMTDH(1) ASRYEVFPLLGPHRMMFKFP(1) TEVWSRFWIGPARSLGVDQS(1) FIGGNYKLGRGRANFVNYAT(1) SQLHWRGPGRTRFVSDRFH(1) AASNEFLGAGRLNFDKPKHS(1) RKDGLIGPGRGGFQGMRWVM(1) GRYLLGPGRELVSLVRPAMH(1) RIVLGPARFFFPQSYLLDQG(1) [YL]-[VLI]-G-P-G-R-x-F 19 Phage display (common panning) 2 PMID:7506691 Anti-HIV-l isotype MN gp120 monoclonal antibody 58.2 Surface protein gp120 of Envelope glycoprotein gp160 Not determined. Not determined. PDL-20 phage display library Micropanning protocol: the mAB was diluted to 50 pg/ml in 0.1M NaHCO3, buffer, pH 9.2, and 50pl added to each well of a Nunc Maxisorb microtiter plate. The mAb was bound to the plate for 2h at room temperature, the wells washed with PBS and blocked with 1% powdered milk in PBS for 1h at room temperature. After blocking, the wells were washed four times with PBS, the phage added in 50pl of milk/PBS and bound to the mAb for 1 h at room temperature. The plates were washed 12 times with 0.5% Tween20 in PBS to remove non-specific phage and the phage were eluted by treatment with 50pl of 0.1M HCl adjusted to pH 2.2 with glycine. Eluted phage were immediately neutralized with 10ul of 1M Tris base. 895 RGDFWQ(2) CRGDCA(1) RGDHWT(1) RRGDLI(1) LRGDLH(1) KRGDLG(1) LKRGDI(1) RCDVW(1) NGRAHA(1) RGD 9 Phage display (subtractive panning) 2-3 PMID:7690752 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. f3-6mer phage display library (X6) Phage were elute with GRGDSP peptide. 896 PKRGDL(3) RGDRLF(1) RGDLFI(1) RRGDLR(1) HRRGDL(1) RGDLHF(1) HRGDLH(1) MRRGDL(1) RGDLFL(1) RGD 9 Phage display (common panning) 2-3 PMID:7690752 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. f3-6mer phage display library (X6) Phage were elute with EDTA after the GRGDSP elution. 897 RGDGWL(5) PKRGDL(4) SRRGDL(4) MRGDLR(1) SLIDIP(1) RGD 5 Phage display (common panning) 2-3 PMID:7690752 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. f3-6mer phage display library (X6) Phage were elute with glycine-HCl buffer of pH 2.2 after the GRGDSP and EDTA elution. 898 CRGDCL(6) RGDRPG(2) TIRSVD(2) RGDSWG(1) RRGDLR(1) KRGDLP(1) PKRGDL(1) KRGDLG(1) RGDLHL(1) RGDFWQ(1) GRRGDI(1) RGDLWT(1) RGDGWL(1) RGD 13 Phage display (common panning) 2-3 PMID:7690752 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. f3-6mer phage display library (X6) The panning was performed by coating with a low concentration (10ng/well) of integrin. Phage were elute with GRGDSP peptide. CRGDCL was 10-fold more efficient than any of the linear RGD-containing hexapeptides in inhibiting experiment. 899 CSRGDGFSC(2) CLSRGDTPC(1) CDRRGDGFC(1) CFTRGDAPC(1) CTSRGDMPC(1) CQLRGDGWC(1) CEGRGDWHC(1) CTLRGDNHC(1) CHLRGDGWC(1) CMLRGDSFC(1) CMPRGDGFC(1) CFRGDHVRC(1) CGRGDSVPC(1) CSRGDGFRC(1) CGRGDNLPC(1) CRGDLRFNC(1) RGD 16 Phage display (common panning) 2 PMID:7507494 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. CX7C phage display library In the first and second panning, the coating concentration of the integrin was 5 microgram/well. 900 CIPRGDGWC(5) CTRGDGWFC(3) CFRGDGFKC(2) CRSRGDFPC(1) CVARGDGWC(1) CFRGDFPEC(1) CRRGDGWEC(1) CRGDWPNYC(1) RGD 8 Phage display (common panning) 3 PMID:7507494 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. CX7C phage display library In the first and second panning, the coating concentration of the integrin was 5 microgram/well. In the third round of panning, the coating concentration of the integrin was 1 microgram/well. 901 CIPRGDGWC(3) CTRGDGWFC(3) CERGDLRMC(2) CYRRGDGHC(1) CELRGDGWC(1) CVARGDGWC(1) CTRGDMQWC(1) CSRGDGWIC(1) CLRGDGFLC(1) CYRGDHLLC(1) CLRGDARFC(1) CTRGDGWPC(1) CGRGDRPQC(1) CKRGDGFWC(1) CRGDFSYMC(1) RGD 15 Phage display (common panning) 3 PMID:7507494 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. CX7C phage display library In the first and second panning, the coating concentration of the integrin was 5 microgram/well. In the third round of panning, the coating concentration of the integrin was 100 nanogram/well. 902 CELRGDGWC(2) CEYRGDGFC(1) CVARGDGWC(1) RGD 3 Phage display (common panning) 4 PMID:7507494 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. CX7C phage display library In the first and second panning, the coating concentration of the integrin was 5 microgram/well. In the third and fourth panning, the concentrations were 100 and 10 ng/well, respectively. 903 CIPRGDGWC(2) CVARGDGWC(1) CQTRGDGWC(1) CLFRGDGWC(1) CLSRGDGWC(1) CFRGDGFVC(1) CRGDGFGSC(1) RGD 7 Phage display (common panning) 5 PMID:7507494 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. CX7C phage display library In the first and second panning, the coating concentration of the integrin was 5 microgram/well. In the fifth round of panning, the coating concentration of the integrin was 10 ng/well. 904 CIPRGDGWC(1) CMTRGDGFC(1) CLFRGDGWC(1) CRGDGFGSC(1) RGD 4 Phage display (common panning) 5 PMID:7507494 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. CX7C phage display library In the first and second panning, the coating concentration of the integrin was 5 microgram/well. In the fifth round of panning, the coating concentration of the integrin was 1 ng/well. 905 CRRETAWAC(15) CASVNGHTC(4) CSTSDVGGC(3) CWANGRSHC(3) CLNTNLGFC(2) CFVNGRSFC(2) CVRLNSLAC(2) CEIVKSSSC(1) CPELFVESC(1) CGPCSGKSC(1) CRGAPRAWC(1) CFAGSLLVC(1) CNLTLSVSC(1) CFANGRHSC(1) CRFGSHVPC(1) CTLVPSRSC(1) CVLNGRMEC(1) CSRPSTFEC(1) CYVNGRVSC(1) CSVANSVVC(1) CWLNGRINC(1) CASFFAVQC(1) CVNVEYRNC(1) CMANGRLLC(1) CHVLASAFC(1) CLNGRGLMC(1) CVFSIAHEC(1) CLVASMTPC(1) CIGTFHHNC(1) CAFYQGLPC(1) CQNAFGYSC(1) CLGEFAFAC(1) 32 Phage display (common panning) 2-5 PMID:7507494 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. CX7C phage display library In the first and second panning, the coating concentration of the integrin was 5 microgram/well. NGR, and its variation, NGH represented alpha5/beta1 integrin binding motif other than RGD. Surprisingly, CRRETAWAC does not bear obvious resemblance to known integrin ligand sequences. However, synthetic cyclic peptide GACRRETAWACGA was a potent inhibitor of integrin alpha5/beta1-mediated cell attachment to fibronectin. This peptide is nearly specific for alpha5/beta1 integrin. CRRETAWAC appears to interact with the same or an overlapping binding site in integrin alpha5/beta1 as RGD. The importance of the disulfide bond was indicated by the fact that the activity of CRRETAWAC peptide was greatly diminished after reduction of the disulfide bond and alkylation of the cysteines. 906 ARLSRK(2) RLSRK(1) LRLSRK(1) GPLRSM(1) RQLRSM(1) LRSM(1) RGMTLS(1) LHMTLS(1) LVAVPT(1) PCSVHT(1) ALAGHS(1) QCGDVK(1) PVLCIT(1) ACCCLN(1) 14 Phage display (common panning) 4 PMID:7523009 Anti-ovarian carcinoma monoclonal antibody OV-TL3 Leukocyte surface antigen CD47 Not determined. Not determined. f3-6mer phage display library (X6) 907 ARLSRK(2) AKLSRK(1) ANLSRK(1) GPFRSM(1) RQLRSM(1) VSRHSM(1) RKLLRT(1) QCVSIM(1) SSPIVS(1) TEPIVS(1) TSLDRI(1) GAYHTV(1) ASCILS(1) SRYIGS(1) 14 Phage display (common panning) 4 PMID:7523009 Anti-ovarian carcinoma monoclonal antibody OV-TL16 Leukocyte surface antigen CD47 Not determined. Not determined. f3-6mer phage display library (X6) 908 SCRGDC(2) DTRGDW(2) CRGDCA(2) FMTRGD(1) CRGDCT(1) SRGDSF(1) NCRGDC(1) SRGDNT(1) FSRGDR(1) LNRGDG(1) WLGRGD(1) DRGDTY(1) RGDSFN(1) NGRIPD(1) TNGRGP(1) NGRSRF(1) TVVRGD(1) RGDAWL(1) RGDSLT(1) TMRGDV(1) VRGDGV(1) TRGDPA(1) RSRNGR(1) NGRNTV(1) RGD 24 Phage display (competitive panning) 3 PMID:7535098 Integrin alpha-V-beta-3, integrin αvβ3 Not determined. Not determined. Not determined. f3-6mer phage display library (X6) Phage were eluted with 0.1mL of 1mM GRGDSP peptide in incubation buffer. Integrin alpha-V beta-3 binds promiscuously to cell-adhesive proteins: vitronectin, fibronectin, and several others containing the RGD motif. 909 SCRGDC(2) SLPRGD(2) WMRGDV(1) GSRGDF(1) NSRGDF(1) FLTRGD(1) AFRGDS(1) VLFRGD(1) KSMRGD(1) GRGDGS(1) PVRRGD(1) MRGDRG(1) SRGDGI(1) RGDSYP(1) VLARGD(1) INSRGD(1) RGDGYV(1) RENRGD(1) GFRGDG(1) TIRGDQ(1) TWNHLS(1) RGD 21 Phage display (common panning) 3 PMID:7535098 Integrin alpha-V-beta-3, integrin αvβ3 Not determined. Not determined. Not determined. f3-6mer phage display library (X6) Phage were eluted with 0.1 M glycine, pH 2.0, containing 1 mg/mL BSA. Integrin alpha-V beta-3 binds promiscuously to cell-adhesive proteins: vitronectin, fibronectin, and several others containing the RGD motif. 910 TWRRGD(2) NGLRGD(2) TKRGDH(1) RRGDHI(1) RGDGSS(1) LQRGDW(1) ATRGDT(1) FRGDFS(1) VLRGDN(1) TRRGDT(1) QIARGD(1) FPVRGD(1) RGDFFS(1) QYLRGD(1) ARWRGD(1) DSARGD(1) RGDSLQ(1) SRLRGD(1) RGDRSL(1) RGDGWL(1) PTQRGD(1) RGDYMD(1) TRGDSL(1) RGDFAF(1) RGD 24 Phage display (competitive panning) 3 PMID:7535098 Integrin alpha-V-beta-3, integrin αvβ3 Not determined. Not determined. Not determined. f3-6mer phage display library (X6) Phage were eluted with low pH in the first round of screening, followed by elution with GRGDSP peptide in the second and third rounds. Integrin alpha-V beta-3 binds promiscuously to cell-adhesive proteins: vitronectin, fibronectin, and several others containing the RGD motif. 911 RGDMSLLGEFTSPYG(8) SRGDGGFIKVLHGSW(3) RGDAIVTFPGSFMLY(2) RRGDIGPRFESAIVD(2) PRGDAFPHMQSSASV(1) PEVIARGDVVFFLRP(1) DALSCRGDCVWPTRG(1) ARGDVFYEGSRGAWY(1) RGDAYAFGNAGVDLI(1) ACGSAGTCSPHLRRP(1) RGD 10 Phage display (competitive panning) 3 PMID:7535098 Integrin alpha-V-beta-3, integrin αvβ3 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Phage were eluted with 0.1mL of 1mM GRGDSP peptide in incubation buffer. Integrin alpha-V beta-3 binds promiscuously to cell-adhesive proteins: vitronectin, fibronectin, and several others containing the RGD motif. 912 RGDAIVTFPGSFMLY(4) GRGDRTDGSSGHVWG(2) GERGDGSFFAFRSPF(2) HAAFEPRGDVRHTLL(2) SRGDGGFIKVLHGSW(2) RGDMSLLGEFTSPYG(2) VRGDSLLFGVQAVLH(1) AGRGDSLGNYRNFNS(1) RGVRGDSFFLVMDAH(1) PRGDAFPHMQSSASV(1) SLRGDHRVRWVLTPH(1) DNSHWFRRISRGDAG(1) MWVFSRGDSSLFCCG(1) RALRGDRGWIVFWDP(1) RGD 14 Phage display (common panning) 3 PMID:7535098 Integrin alpha-V-beta-3, integrin αvβ3 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Phage were eluted with 0.1 M glycine, pH 2.0, containing 1 mg/mL BSA. Integrin alpha-V beta-3 binds promiscuously to cell-adhesive proteins: vitronectin, fibronectin, and several others containing the RGD motif. 913 RGDMSLLGEFTSPYG(10)[NT] DALSCRGDCVWPTRG(6)[100] RRGDIGPREFSAIVD(4)[NT] SPARGDLFRFMGAVH(2)[NT] PRDHIARRGDLAFQA(1)[NT] RGD 5 Phage display (competitive panning) 3 PMID:7535098 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. f3-15mer phage display library (X15) Binding assay IC50 (nM) values for the inhibition of binding of adhesive proteins to αvβ3 or α5βl integrins by synthetic peptides were measured using a solid-phase assay. Phage were eluted with 0.1mL of 1mM GRGDSP peptide in incubation buffer. 914 RRGDIGPREFSAIVD(5)[NT] ISRRGDLSGLSFSRL(4)[4000] RGDMSLLGEFTSPYG(4)[NT] DALSCRGDCVWPTRG(3)[100] RGVKMRRGDFSTIMD(1)[NT] SPARGDLFRFMGAVH(1)[NT] RVFRGDLGYRTPYIG(1)[NT] RGDVWTLWSVGDTRS(1)[NT] SEELLVESSAIRSRE(1)[NT] RGD 9 Phage display (common panning) 3 PMID:7535098 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. f3-15mer phage display library (X15) Binding assay IC50 (nM) values for the inhibition of binding of adhesive proteins to α5β1 integrins by synthetic peptides were measured using a solid-phase assay. Phage were eluted with 0.1 M glycine, pH 2.0, containing 1 mg/mL BSA. 915 RETADDLLSLLL VRYPSDLLLMIL STEPDLLWIIMG QDDILWEILAVD LGNNDDLLFFNL RGCGVDLLDCLL D-L(2)-x(2)-L(2) 6 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE14 Proenkephalin-A Not determined. Not determined. X12 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 916 HDDSDLLLTFLR RETADDLLSLLL EYTNDLLLRLLS STEPDLLWIIMS IDTDLLYVITQS TADFLYLITTWS TSDLGMYLFGSV L(2)-x(2)-L 7 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE15 Proenkephalin-A Not determined. Not determined. X12 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 917 RETADDLLSLLL EYTNDLLLRLLS D-L(2)-x(2)-L(2) 2 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE19 Proenkephalin-A Not determined. Not determined. X12 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 918 HDDSDLLLTFLR EYTNDLLLRLLS D-L(2)-x(2)-L 2 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE23 Proenkephalin-A Not determined. Not determined. X12 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 919 RETADDLLSLLL EYTNDLLLRLLS LDSDLQLFFLSS 3 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE25 Proenkephalin-A Not determined. Not determined. X12 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 920 GDVTWPQGEEWTEEHWVRWS DNGLAGRDYERDVRERVMKL DPRVWGRDVEPHVFYKYWRN QTSDSLGGKDDESWHTTAKL QEEFVHGMDRERVVWLLTSA G-x-D-x-E-x(2)-V 5 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE13 Proenkephalin-A Not determined. Not determined. X20 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 921 LGAENSGGQDRSDVTAARYG AGRMSQRGVDQVEWHLSRFY SVPAGADQVSFHTDRALGAV AGEDSGAHHRLRWETGRARW 4 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE20 Proenkephalin-A Not determined. Not determined. X20 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 922 SVPAGADQVSFHTDRALGAV ERTSGVDNPRYHATWTTRLR HPQGEDVQKSVVARWGDFLR VKGELRGVDERTRVMQQYG RDGKEGTQEAMYVMERYSIL AGEDSGAHHRLRWETGRARW TWKDRSTGGGMDMPGWHLQF 7 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE21 Proenkephalin-A Not determined. Not determined. X20 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 923 HPQGEDVQKSVVARWGDFLR GYGEDRERETWARWDRVTQA VAQGMAGQDKEREFRERLQN SVPAGADQVSFHTDRALGAV GLVLWGEDREEWHQSLQAGL THSAKTSLEGIDREAMVFIW KDMGRYGEDREHKVREWMRQ 7 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE22 Proenkephalin-A Not determined. Not determined. X20 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 924 MFGEDNYEFHFR MRMGYDNEKYHT G-x-D-N 2 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE24 Proenkephalin-A Not determined. Not determined. X12 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 925 SKLSPAMPGEDKERELAAKL VASIGGKDNEEAVWEHRGAV EPTGRDAEDWIYRAQGRFYL DLGVDIEMEVASRVRGGHSY VAQGMAGQDKEREFRERLQN WPYSCGQDNTSKVANRYARF HPQGEDVQKSVVARWGDFLR G-x-D-N 7 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE24 Proenkephalin-A Not determined. Not determined. X20 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 926 DLLFEL DLLWDL DLLYDL DLLYHL DLLWLV DLFWEL DLMFIM D-L(2)-x(2)-L 7 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE14 Proenkephalin-A Not determined. Not determined. f3-6mer phage display library (X6) Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 927 DLLWSL DLLYEL DLLWHL DLLYDL DLMWAL DLLWES DLLFYY DWLFDR DRLWHY DWLWHY DFMIEL 11 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE15 Proenkephalin-A Not determined. Not determined. f3-6mer phage display library (X6) Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 928 GGWMRGHTHLST TDGGGWMVGYRM KFQSPIAWGFVR GMTILGGRMGFR 4 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE1 Proenkephalin-A Not determined. Not determined. X12 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 929 GPEEEAGGGGFMRGASRMTQ KLAGGWMQGHTVVPSREAKA GGFMAGIREKGAGREWNALT TVGGWMWGQGLKMAGQGRQS MKPSCAHADHGALGWMRGRL TEDRDLPEVLGGWMRGRYPN EGWMRGYRGFLQSQNLESGW G(2)-W-M-R-G 7 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE1 Proenkephalin-A Not determined. Not determined. X20 fAFF1 phage display library Three rounds of biopanning were carried out with decreased antibody concentrations (1 mg/ml and 0.1 mg/ml). 930 IWTWEPSKRYQL 1 Phage display (common panning) 3 PMID:7536850 Anti-proenkephalin monoclonal antibody PE2 Proenkephalin-A Not determined. Not determined. X12 fAFF1 phage display library For antibody PE2, the amplified library after the first round was subjected to two further rounds of antibody panning. In each round, 10 ml of library was panned on Petri dishes coated with 10 mg/ml PE2 and subsequently blocked with TBST buffer containing 5% (w/v) low-fat milk powder, for one hour. 931 YGSWAS(2)[NT] YGGLGL(1)[NT] YGSLVL(1)[NT] YGGLGI(1)[NT] YGSLVQ(1)[NT] YGGLGR(1)[NT] YGSLVR(1)[NT] YGGLNV(1)[NT] YGSLAD(1)[NT] YGGLRA(1)[NT] YGSLLS(1)[NT] YGGLEM(1)[NT] YGSLNG(1)[NT] YGSLYE(1)[NT] YGGIAS(1)[NT] YGGIAV(1)[NT] YGGIRP(1)[NT] YGSWQA(1)[NT] YGGWAG(1)[7.8] YGGWGP(1)[NT] YGSFLH(1)[NT] YGGWSS(1)[NT] YGGMKV(1)[NT] YGGFPD(1)[2.3] YGALGG(1)[NT] YGALSW(1)[NT] YGALDT(1)[NT] YGALEL(1)[NT] YGAIGF(1)[NT] YGAWTR(1)[NT] YGWWGL(1)[NT] YGLWQS(1)[NT] YGWWLT(1)[NT] YGFWGM(1)[0.35] YGWLAT(1)[NT] YGKWSG(1)[NT] YGWANK(1)[NT] YGPFWS(1)[1.9] YGEFVL(1)[NT] YGNWTY(1)[7.8] YGDFAF(1)[NT] YGNFAD(1)[NT] YGNFPA(1)[NT] YAWGWG(1)[NT] YAGFAQ(1)[8.3] YGTFIL(1)[NT] YGTWST(1)[NT] YSMFKE(1)[NT] YGVWAS(1)[NT] YGVWWR(1)[NT] 52 Phage display (common panning) 3 PMID:2201029 Anti-beta-endorphin monoclonal antibody 3-E7 Beta-endorphin Not determined. Not determined. X6 fAFF1 phage display library Radioimmunoassay A solution radioimmunoassay (RIA) was used to estimate the binding affinities of peptides for mAb 3-E7. Six hexapeptides with sequences corresponding to those of antibody-selected phage were chemically synthesized and assayed for binding to 3-E7. Each Kd (μM) value is the antilogarithm of the mean of log Kd from three to six independent determinations with standard deviations of <0.25 logarithm units in all cases. NT denotes not tested. Compared with 7.1 nM for a known high-affinity ligand (YGGFL), the binding affinities of six chemically synthesized hexapeptides from this set range from 0.35uM (YGFWGM) to 8.3uM (YAGFAQ). 932 WNVHGIWQE(2) DWDTRGLWVA(2) WWTDTGLW(1) WWTDDGLW(1) WWDTRGLWVWTI(1) FWGNDGIWLESG(1) DWDQFGLWRGAA(1) RWDDNGLWVVVL(1) CWSMHGLWLC(1) SGMWSHYGIWMG(1) GGRWDQAGLWVA(1) KLWSEQGIWMGE(1) GCWDNTGIWVPC(1) DWDTRGLWVY(1) SLWDENGAWI(1) KWDDRGLWMH(1) QAWNERGLWT(1) QWDTRGLWVA(1) SWDTRGLWVE(1) SWGRDGLWIE(1) EWTDNGLWAL(1) SWDEKGLWSA(1) SWDSSGLWMD(1) W-x(3)-G-x-W 23 Phage display (common panning) 3-4 PMID:8693002 Interleukin-1 receptor type 1 Not determined. Not determined. Not determined. X8, X10 and X12 phage display library pool Extracellular domain of type I IL-1 receptor was fuse to Fc domain of IgG1. The IL-lR-Fc fusion was then used as target. These peptides blocked binding of IL-la to the type I IL-1 receptor with IC50 values of 45-140uM. They also blocked IL-1-driven responses in human and monkey cells; howver, they did not bind the human type II IL-1 receptor or the murine type I IL-1 receptor. 933 HITWDQLWNVMN 1 Phage display (common panning) 4 PMID:7545665 E-selectin Not determined. Not determined. Not determined. X12 fAFF1 phage display library The soluble extracellular domain of E-selectin was expressed in a phosphatidylinositol (PI)-glycan-linked form on Chinese hamster ovary cells and was released from the cell surface with PI-PLC. A monoclonal antibody, mAb179, recognizes a portion of the PI-glycan linkage signal sequence at the C terminus of the released receptor and was used to immobilize active receptor in microtiter wells for panning. 934 LRHSVI(5) RHSVIS(1) FRHSLL(1) WRHSVV(1) FKHSVV(1) LRHSIL(1) YRHSIV(1) RHS 7 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody PAb240 Cellular tumor antigen p53 Not determined. Not determined. f3-6mer phage display library (X6) First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 935 EGRHSVVHQGAA(2) FRHSVVDYGLYN(1) YSRHSVVYGDGQ(1) TLRHSIIFGGEW(1) AVRHSVIERTLS(1) GPGIRHSTVPAY(1) DRAPRIRHSVII(1) WIATTLMRHSVV(1) RHS 8 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody PAb240 Cellular tumor antigen p53 Not determined. Not determined. X12 fAFF1 phage display library First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 936 DGEQGFMINERHSVIPPWSS(4) DRHSVVTEDPGALDRRTHSF(1) DGALEERHSVLRRTEAEYQM(1) TQASAELERHSVVIRQPRFV(1) EPLEKCVRRHSMRDLSGVCP(1) TEVGNMASRVRHSVIEAANG(1) IELGEPKKVVVRHSIIGSHL(1) SGHHSEMRALKSSLRHSVIS(1) RHS 8 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody PAb240 Cellular tumor antigen p53 Not determined. Not determined. X20 fAFF1 phage display library First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 937 SDLYKL(6) SDLHKL(5) SDLNKL(1) S-D-L-x-K-L 3 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody DO-1 Cellular tumor antigen p53 Not determined. Not determined. f3-6mer phage display library (X6) First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 938 GWSDLHKLPPHT(3) DPQSDLGKLRTG(2) TGAPNSDLFKLS(2) YSDLWKLYRFDA(1) GGASDLDRLLSE(1) NMQSDAWKLLHN(1) WLASDLDKLVLG(1) NEVPWRGFQDPN(1) S-D-L-x-[KR]-L 8 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody DO-1 Cellular tumor antigen p53 Not determined. Not determined. X12 fAFF1 phage display library First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 939 SDLNKL(6) SDLHKL(4) SDLFKL(2) S-D-L-x-K-L 3 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody DO-7 Cellular tumor antigen p53 Not determined. Not determined. f3-6mer phage display library (X6) First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 940 GWSDLHKLPPHT(5) DPQSDLGKLRTG(3) GGASDLDRLLSE(2) YSDLWKLYRFDA(1) LEYSDLDRMTWP(1) S-D-L-x-[KR]-[LM] 5 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody DO-7 Cellular tumor antigen p53 Not determined. Not determined. X12 fAFF1 phage display library First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 941 SDLHKL(4) FSDLHK(2) FSDLNK(2) SDLWRL(1) SDLYKL(1) SDLGKL(1) FSDLYK(1) S-D-L-x-[KR] 7 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody Bp53-11 Cellular tumor antigen p53 Not determined. Not determined. f3-6mer phage display library (X6) First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 942 TGAPNSDLFKLS(5) DPQSDLGKLRTG(4) GWSDLHKLPPHT(1) GGASDLDRLLSE(1) NDTYSDLTRMIH(1) S-D-L-x-[KR]-[LM] 5 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody Bp53-11 Cellular tumor antigen p53 Not determined. Not determined. X12 fAFF1 phage display library First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 943 SDLHKL(13) 1 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody Bp53-12 Cellular tumor antigen p53 Not determined. Not determined. f3-6mer phage display library (X6) First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 944 YSDLWKLYRFDA(4) GWSDLHKLPPHT(4) HFSSRFSDLGKM(4) LTTHMSDLDRMH(1) S-D-L-x-[KR]-[LM] 4 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody Bp53-12 Cellular tumor antigen p53 Not determined. Not determined. X12 fAFF1 phage display library First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 945 DLWSWR(2) FDLWSF(2) DLWALL(1) DLWAFD(1) DLWSLL(1) DLWHFP(1) DLWRLV(1) DLWRFS(1) PDLWRF(1) LPDLWR(1) DLW 10 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody Bp53-19 Cellular tumor antigen p53 Not determined. Not determined. f3-6mer phage display library (X6) First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 946 SPTTVADLWAIM(3) FGDLWCFACNAV(1) LNDLWSISDPEW(1) ALTDLWSFLVLE(1) TTRLVDLWGFHL(1) TFHFSTDLWRLA(1) DYGRAMADLWQF(1) SNPNYPGDLWMF(1) RAHTGIMDLWGF(1) IARSAIHDLWSL(1) DLW 10 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody Bp53-19 Cellular tumor antigen p53 Not determined. Not determined. X12 fAFF1 phage display library First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 947 NKMWGK(5) RMNRPK(3) RFHAKF(3) RFWAKF(2) NRMWGK(1) 5 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody PAb421 Cellular tumor antigen p53 Not determined. Not determined. f3-6mer phage display library (X6) First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 948 KYLYSTTSSHVV(5) EWKGMNFIGSRP(4) TTSWHGRPDRNL(3) ITSHHGRPRALI(2) KEAASTSSADRR(2) LWVPSLFFSKLS(1) GDTWHGAYFYGR(1) KTNNTVCSKSVC(1) 8 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody PAb421 Cellular tumor antigen p53 Not determined. Not determined. X12 fAFF1 phage display library First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 949 GNGAVWEGKKLSTSSSLRVL(4) TKGKVDTGTIFGNAPRMAIS(2) GCGTSWQGELCKSIRLRYVP(2) APTGAKKGGTTSSLASHYRN(2) VHGEMVRKSFNSTTSRVPRT(1) WTESGRLTKRESSTNKNLGR(1) 6 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody PAb421 Cellular tumor antigen p53 Not determined. Not determined. X20 fAFF1 phage display library First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 950 KYSDSVSLTHRI(10) KLSQSTSPPVTG(1) GIKLGDSVSVWR(1) 3 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody PAb122 Cellular tumor antigen p53 Not determined. Not determined. X12 fAFF1 phage display library First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 951 YLHQPGRIDCTARPLEKCSR(7) KSEGKAEAVWAFKAHGSAVS(3) VGQTKPKVEKLGESTSMRLL(2) GSESTAERKSPDTPQLVEKL(1) MAARWVQPMPFSKWTESLRG(1) 5 Phage display (common panning) 3 PMID:7537340 Anti-P53 monoclonal antibody PAb122 Cellular tumor antigen p53 Not determined. Not determined. X20 fAFF1 phage display library First round, 10000ng of monoclonal antibody; second round, 100ng; third round, 10ng. 952 TTRSPRLPIELLFAG(4) LHTPKPPWSVTSFVP(3) VSRIPISYLLQYSNR(2) TLTNVRLSLDSWLKR(2) SYPKLPWSFIKQLNK(1) SLSTEPRLPWSFFFS(1) MQISLSPRIPWSKIL(1) PTVLIRKSLPWSLIL(1) TLWTSTRFPIPWSLL(1) PQTPWGYLWDTSRFR(1) SMLEPRSKISWKSIL(1) NLSKKLSWDRVLTMM(1) HAKLNWDYILNTANV(1) LSLPKKLAWADILPS(1) LSQANRALLSWDKIL(1) LFSSKVPISYLFDIT(1) NLKLPLTFLDLSRLY(1) RIPLFNLSDLMGMQS(1) TTQRLPMRIPEVFSF(1) LPAKLPMESWLRSHP(1) KIPAQLWMQFAKSTN(1) PYWSNRAKLPMQWYQ(1) LKSPLTWDQVLLQSS(1) TYPPRLSTEFLFGNV(1) VTSALHQRLNISHIF(1) LSSVKLTLRQLFPYV(1) MPRGRLKVSEIFSTR(1) LNPGYLRKLNVLSIL(1) QWLLMKANPLDLLYR(1) LPMMKRDLRNLLNIV(1) STVTRMDLLKLLSSM(1) PVKLHMDILEVLQSH(1) PTPKQKINLQDWLLL(1) KLSIWDVLFDQAFAG(1) ATQHRKYLTLDDWLR(1) SPPWVFREHVYNLLP(1) DVPLNWSTFFSPSTR(1) AYHWSQSPLAIERIL(1) RHHTLIDLMSLLALQ(1) RLLSWSQILTLSDST(1) YPLPIMRLLDSFSSK(1) LLNMSRVLEYHTPSK(1) MQQTNSWMKWKTQAF(1) AFPHAPMSIDHIFFT(1) [KR]-[LI]-x-W-x(2)-I-L 44 Phage display (common panning) 3-5 PMID:7540176 Protein S100-B F-actin-capping protein subunit alpha-1 Not determined. Not determined. X15 M13 phage display library S-100b binding bacteriophages were selected by Ca(2+)-dependent affinity chromatography, and the sequence of the random peptide insert contained in 51 clones was determined. 953 EHDFHHIREWGNHWKNFLAVM[150 ± 30] YEKPWQNLWDWGAEAFKDLLDK[240 ± 80] LSFDWSELRRWGTWAAAEVFEL[150 ± 40] ELWRNLRLWGYCMNLSNMPL[930 ± 130] EFSLFKDIYRWGNWAAGFYYGV[NT] SFCLFDSIFDWGARGAEWHVG[NT] SCNMWRNIYSWGATFPQDHIFF[NT] DMDLFQHLSCWGTDFADMMFDE[NT] SCAWFLHLRQWGNSNWTYIFGV[NT] MHFGWYHLSEWGKIAALIGFEM[NT] ETGWFHNLTAWGEESVYALHIM[NT] SPGLFDNLKTWGTRMEHFMSLS[NT] QSFSMMSRWGQEFHNMLINV[NT] TALGWVHMHEWGEEFSKFPWGN[NT] QDYWTNMLIWGSMLLWSLFGADP[NT] HDYWQSMNHWGGVYGGCVPMCK[NT] YEEMWQWVFKAGQEWPWSYFPM[NT] QIPFQIQAYWLDVCCQEVTDDL[NT] IDQMWQWDPRWGXXXEEIDIHD[NT] SAFEYIQLTVSGWLPHLMGLF[NT] EKFEILCINYQGWISDLCEAIA[NT] WHDEDVLFRNLGEATLWLAEYY[NT] DWECMYDFCWRGXCVALNSTG[NT] DQTYKFEYNMGKDVTSLVCYW[NT] DNWIDSLVCIVGTGYGSLAAYY[NT] [WF]-x-[LIM]-x-W 25 Phage display (common panning) PMID:7542403 Antigen-presenting glycoprotein CD1d1 Not determined. Not determined. Not determined. X11GX12 phage display library Binding assay Dissociation constants (nM) for unlabeled peptides were determined from the molar concentration giving 50% inhibition of 125I-p99 binding to mCD1 in a competitive binding assay. NT denotes not tested. The recombinant soluble CD1-beta2 microglobulin complexes (mCD1) expressed in Drosophila melanogaster SC2 cells were used in the panning. The mCD1 was also engineered with a COOH-terminal hemagglutinin (HA) tag (YPYDVPDYAS), an epitope derived from the influenza HA protein. In this way, mCDl-phage complexes could be identified with a HA tag-specific antibody. Forty-seven different clones were selected by mCD1 binding. However, only 25 sequences shows a well-defined core motif were given in the original paper. 954 AGELSMQEWQRFRDLSLGRS RTPPLAETPEWQYQRYLFLL 2 Phage display (common panning) 3 PMID:7543411 Anti-keratin 19 monoclonal antibody BM19.21 Keratin, type I cytoskeletal 19 Not determined. Not determined. X20 fAFF1 phage display library At least 10 individual phage clones were sequenced. Only unique sequences were given. 955 MWDERWEQEELLGRRLLEAP 1 Phage display (common panning) 3 PMID:7543411 Anti-keratin 19 monoclonal antibody LE64 Keratin, type I cytoskeletal 19 Not determined. Not determined. X20 fAFF1 phage display library At least 10 individual phage clones were sequenced. Only unique sequences were given. 956 GESMAERSMAQERLLTYFMV GGGERSLVQESLLWSASSYA EADRAADQEAQMWTTKAVML ESDRDWGQECSNWYADCRYP [ED]-R-x(3)-Q-E 4 Phage display (common panning) 3 PMID:7543411 Anti-keratin 19 monoclonal antibody KM4.62 Keratin, type I cytoskeletal 19 Not determined. Not determined. X20 fAFF1 phage display library At least 10 individual phage clones were sequenced. Only unique sequences were given. 957 NEWGSKDELEFYSHLVSSLR EQAELTRTMQQLFRGTQVRS GEVMHQGDWGAENFYLTLSA AREGKDDQLFLYQSLQRYWR EAWDKRDEQTIYESLSWLSA LGEDKERHRESWLVYKTLTW GENAEALEAQVYYQSLRDFW QGWDHLYGPYYFTAADEGNL 8 Phage display (common panning) 3 PMID:7543411 Anti-keratin 19 monoclonal antibody LP2K Keratin, type I cytoskeletal 19 Not determined. Not determined. X20 fAFF1 phage display library At least 10 individual phage clones were sequenced. Only unique sequences were given. 958 AIVIENYYTLCC HLQIYYTSLA 2 Phage display (common panning) 3 PMID:7543411 Anti-keratin 19 monoclonal antibody LP2K Keratin, type I cytoskeletal 19 Not determined. Not determined. X12 fAFF1 phage display library At least 10 individual phage clones were sequenced. Only unique sequences were given. 959 SRGDYLSWEAYKTLMGASSS VSWEAYGEGAAGFFRLAEAL SWEAY 2 Phage display (common panning) 3 PMID:7543411 Anti-keratin 19 monoclonal antibody BA16 Keratin, type I cytoskeletal 19 Not determined. Not determined. X20 fAFF1 phage display library At least 10 individual phage clones were sequenced. Only unique sequences were given. 960 CRQNWGLEGC(26)[9548] CVDVVSASTC(4)[5720] CVRSESMLHC(3)[176] CRDPRAQDLC(1)[89] CLRSGIQNDC(1)[79] CAHLRAGRQC(1)[76] CSTMPPFASC(1)[64] CMDRGISQAC(1)[64] CLSAMVSGRC(1)[46] CPNSSGIRHC(1)[68] CWSLSTSIDC(1)[46] CSGGGFGSYC(1)[49] 12 Phage display (common panning) 7 PMID:7559473 MAR DNA Affinity Column DNA-binding protein SATB1 Not determined. Not determined. CX9 phage display library Binding assay The binding of selected phage clones to MAR was determined by a filter binding assay. The level of binding was quantitated by counting each spot in a liquid scintillation counter. Phage Selection were combined procedures for biopanning and methods for purifying DNA-binding proteins. One predominant cyclic peptide CRQNWGLEGC selected by a MAR-affinity column showed 50% identity with a segment in SATB1 (amino acids 355-363). 961 KPWYVSRV GKPWWASR EKPWWAVR GKPWYAGR KPSNVSRV YYCSPWCD VPWYKQST APWYRVSP LPWYLYPS 9 Phage display (common panning) 4 PMID:8170958 Purified IgM lamda receptor of the human Burkitt lymphoma cell line SUP-B8 Not determined. Not determined. Not determined. 8-mer ASASA-pIII phage display library All the peptides bound to the SUP-B8 immunoglobulin specifically. Their binding could be inhibited with anti-Id antibodies but not with anti-IgM or anti-lamda light-chain antisera. When conjugated to form dimers or tetramers, they induced cell death by apoptosis in vitro with an IC5o between 40 and 200 nM. This effect was associated with specific stimulation of intracellular protein tyrosine phosphorylation. 962 KPWWVSRV[0.97] KPWYVGRP[NT] KPWWVTRV[NT] KPWWVVRL[NT] GKPIWAGR[NT] QKPIWVTR[NT] DWAVWNRR[NT] NWAVWTKR[NT] NWGMWSKR[NT] YVFEDLFR[NT] MPEDFYRR[NT] SEPVDHGL[NT] VDPVDHGL[NT] VPIDHGT[NT] 14 Phage display (common panning) 4 PMID:8170958 Purified IgM lamda receptor of the human Burkitt lymphoma cell line SUP-B8 Not determined. Not determined. Not determined. 8-mer P4-pIll and 8-mer P6-pIII phage display library ELISA Biotinylated peptide monomer (0.4 μg per well) was immobilized on a microtiter well coated with 1 μg of streptavidin per well. SUP-B8 immunoglobulin or control immunoglobulin (0.25 μg) was added. Optical density values, OD (405-490 nm), were measured. Data shown were reproduced from the graph and expressed as means of duplicate samples ± SD. All the peptides bound to the SUP-B8 immunoglobulin specifically. Their binding could be inhibited with anti-Id antibodies but not with anti-IgM or anti-lamda light-chain antisera. When conjugated to form dimers or tetramers, they induced cell death by apoptosis in vitro with an IC5o between 40 and 200 nM. This effect was associated with specific stimulation of intracellular protein tyrosine phosphorylation. 963 KNGPWYAYTGRD FGILTEEMYRRW LRYTQEEMYRRW HYVHEDLYRRVK VTGYTMDVLYRR 5 Phage display (common panning) 4 PMID:8170958 Purified IgM lamda receptor of the human Burkitt lymphoma cell line SUP-B8 Not determined. Not determined. Not determined. 12-mer GG-pIII phage display library All the peptides bound to the SUP-B8 immunoglobulin specifically. Their binding could be inhibited with anti-Id antibodies but not with anti-IgM or anti-lamda light-chain antisera. When conjugated to form dimers or tetramers, they induced cell death by apoptosis in vitro with an IC5o between 40 and 200 nM. This effect was associated with specific stimulation of intracellular protein tyrosine phosphorylation. 964 PQVRPI(17) WIVNSP(1) PRVQIL(1) 3 Phage display (common panning) 3 PMID:7622517 Anti-p22phox monoclonal antibody 44.1 Cytochrome b-245 light chain Not determined. Not determined. f3-6mer phage display library (X6) Western blotting demonstrated specific binding of the antibody to the selected phage peptides. 965 HDQPMVLPI(8) EGRFGGGQV(3) AQPQVRPIG(1) NMPQVRPID(1) DRPRVKPK(1) GTPEVRPV(1) QPVVRPIDA(1) KEKPMVRPL(1) KPMVRPLVN(1) RPQVIPISV(1) KPMVIPIHS(1) QQPRVLPLD(1) MTPEVRPID(1) ENRPKVSPI(1) NKPRVMPLD(1) TGPEVRPLQ(1) RPMVHPLTP(1) DRPWVRPIV(1) TTRPEVRPI(1) HHHPMIPPI(1) EDRPMVRV(1) 21 Phage display (common panning) 3 PMID:7622517 Anti-p22phox monoclonal antibody 44.1 Cytochrome b-245 light chain Not determined. Not determined. X9 phage display library Western blotting demonstrated specific binding of the antibody to the selected phage peptides. 966 PKGAYD(9) FKRGVD(5) LREGVD(1) PRPAVD(1) LRRGID(1) 5 Phage display (common panning) 3 PMID:7622517 Anti-gp91phox monoclonal antibody 54.1 Cytochrome b-245 heavy chain Not determined. Not determined. f3-6mer phage display library (X6) Western blotting demonstrated specific binding of the antibody to the selected phage peptides. 967 GRPPKAGVD(3) GGNLKVGVD(1) KFYPKQSVD(1) QKLSKLAVD(1) NRLDKAAVD(1) IARNAVDGP(1) KAVEKSAVD(1) DAMPAEGVD(1) KTILKQGVD(1) SGPPKPALD(1) SSIIKDGVD(1) EMPLKRGVD(1) ERMPAHGVD(1) SRPPKGAID(1) KIPPRSAVD(1) SKTPKEGLD(1) 16 Phage display (common panning) 3 PMID:7622517 Anti-gp91phox monoclonal antibody 54.1 Cytochrome b-245 heavy chain Not determined. Not determined. X9 phage display library Western blotting demonstrated specific binding of the antibody to the selected phage peptides. 968 PWWWVSWVDAGGGSLALPTQPSD WGWPTWWGWTGGDARHPSAPEAH WWDPDIWFGWGGAHPPNLIQPIS GWQSGWEWWIGGGNWTSNTTH PYWMFYGFDWRGGFPPSHQIMDQ WMQSWYYHWGGGETFPIRRDSGG 6 Phage display (common panning) 3 PMID:7737512 96-well immunoassay plate (polystyrene, PS) Not determined. Not determined. Not determined. TSAR-12 phage display library The plastic-binding phage bind to both unblocked plastic (polystyrene, PS and polyvinyl chloride, PVC) and plastic blocked with bovine serum albumin (BSA) but require non-ionic detergent to bind to plastic blocked with milk. 969 EIHGNLYNWSPLLGYSYFPGISPKHISGEVPPRPSTKV YTGWETWYSFDPFTHYGGPGSRFDFVHDKSEDPIDRSY QDLDHWSYWSMYSTYPTSPGLVPYSWGYGSPNSHTDKL IYYPFFVWGNYANGGLLSPGHVYSSNFIPLYMQREVSP SSRLAYDHYFPSWRSYIFPGSNSSYYNNSWPTITMETN RPYLYDPNEWHRYYSYLLPGHSYNVQSWPDGLGKSASR LGFSGWYWQGLYGLGSHDPGFIHEQSPAEVAMEDTEQS QTLIDFHDLHYWGAYYGWPGIYDEASGSQAVRHNMTHT DSWPLRIYSGLSNYYHYFPGSLVYNMMYPSLSRLPKGD HHGAMNRYYTWLWDNSRFPGRSYLLSAPATQPEASISQ TYDYTYDWSGLFWSPFTHPGAHMTTHSPWAGHKPHAET 11 Phage display (common panning) 3 PMID:7737512 96-well immunoassay plate (polystyrene, PS) Not determined. Not determined. Not determined. TSAR-9 phage display library (X18PGX18) The plastic-binding phage bind to both unblocked plastic (polystyrene, PS and polyvinyl chloride, PVC) and plastic blocked with bovine serum albumin (BSA) but require non-ionic detergent to bind to plastic blocked with milk. 970 RPHQVV(1) LPFNMT(1) VPRAIS(1) EAFGMR(1) RPQTLA(1) 5 Phage display (common panning) 3 PMID:7896777 Stromelysin-1, SL-1 Not determined. Not determined. Not determined. X6 fAFF1-tether C phage display library In the first round, 2E11 TU (about 1000 equivalents of each clone) were treated with stromelysin. The output was not titered before the next round of screening, thus variable amounts of phage were used as input for the succeeding round. In later rounds of screening, the protease concentration was reduced. After each round, 12 clones were analyzed using the phage proteolysis assay. 971 GPMAMT(1) APQATV(1) APSAMS(1) 3 Phage display (common panning) 4 PMID:7896777 Stromelysin-1, SL-1 Not determined. Not determined. Not determined. X6 fAFF1-tether C phage display library In the first round, 2E11 TU (about 1000 equivalents of each clone) were treated with stromelysin. The output was not titered before the next round of screening, thus variable amounts of phage were used as input for the succeeding round. In later rounds of screening, the protease concentration was reduced. After each round, 12 clones were analyzed using the phage proteolysis assay. 972 IPFEQR(1) 1 Phage display (common panning) 5 PMID:7896777 Stromelysin-1, SL-1 Not determined. Not determined. Not determined. X6 fAFF1-tether C phage display library In the first round, 2E11 TU (about 1000 equivalents of each clone) were treated with stromelysin. The output was not titered before the next round of screening, thus variable amounts of phage were used as input for the succeeding round. In later rounds of screening, the protease concentration was reduced. After each round, 12 clones were analyzed using the phage proteolysis assay. 973 RPHQVV(1) PFNSVS(1) KAFELT(1) SLYELR(1) 4 Phage display (common panning) 6 PMID:7896777 Stromelysin-1, SL-1 Not determined. Not determined. Not determined. X6 fAFF1-tether C phage display library In the first round, 2E11 TU (about 1000 equivalents of each clone) were treated with stromelysin. The output was not titered before the next round of screening, thus variable amounts of phage were used as input for the succeeding round. In later rounds of screening, the protease concentration was reduced. After each round, 12 clones were analyzed using the phage proteolysis assay. 974 VPMSMS(2) GANFFE(1) NDYPAF(1) GVDFLE(1) FNEFSN(1) RPESMT(1) APLQST(1) IPSEMR(1) KPHALS(1) APLESI(1) APLEAV(1) APLTLS(1) EPMSLS(1) PKPLAL(1) PMAMMG(1) APFRLE(1) VPLETR(1) GPYSLT(1) VPMEIK(1) AWLLRW(1) EPYELY(1) TPFAFA(1) VPYELA(1) EPLGLV(1) APYEVM(1) MLLAST(1) GPWESK(1) PMEMVE(1) VPLELK(1) PMAMVA(1) 30 Phage display (common panning) 4 PMID:7896777 Stromelysin-1, SL-1 Not determined. Not determined. Not determined. X6 fAFF1-tether C phage display library In the first round, 2E10 TU (about 100 equivalents of each clone) were treated with stromelysin at 1 μg/ml. The output phage were titered before the next round, and a constant input was used for the succeeding round. 975 EPMSLS(2) MPLSLT(1) PLEIRA(1) HPMDVK(1) MLAELR(1) ILELQG(1) APMQLE(1) MILDLK(1) EPMDLI(1) VMELQG(1) YAMELR(1) SIQALT(1) RPLQIV(1) PANIKG(1) VPMNMT(1) AKKARM(1) PMTLAG(1) IPLPLT(1) KKVRRV(1) RRVRKV(1) GPLGLH(1) 21 Phage display (common panning) 3 PMID:7896777 Matrilysin Not determined. Not determined. Not determined. X6 fAFF1-tether C phage display library 976 PMPHSLNFSQYLWYT(11)[0.01] MHRSLWEWYVPNQSA(9)[>23] HTYSSLWDTYSPLAF(8)[0.34] SQTGTLNTLFWNTLR(8)[10.0] WHPGLSFGSYLWSKT(6)[0.40] SSLWTRYAWPSMPSY(5)[0.40] TLFMDLWHDKHILLT(4)[8.0] ILNFPLWHEPLWSTE(2)[9.0] WSFYNLHLPEPQTIF(2)[1.0] PLDLWSLYSLPPLAM(2)[2.0] LDLWMRHYPLSFSNR(1)[0.38] PALLNWSFFFNPGLH(1)[1.0] PTLWQLYQFPLRLSG(1)[2.5] ISFSELMWLRSTPAF(1)[5.0] LSEADLWTTWFGMGS(1)[7.0] SSLWRIFSPSALMMS(1)[8.0] SLPTLTSILWGKESV(1)[8.0] IKTDEKMGLWDLYSM(1)[23.0] SHIKSLLDSSTWFLP(1)[>47] 19 Phage display (subtractive panning) 3 PMID:8041758 Urokinase plasminogen activator surface receptor Urokinase-type plasminogen activator (uPA) 2FD6,2I9B,3BT1,3BT2,3U73,4K24, Not determined. X15 M13 phage display library Binding assay The IC50 (μM) was determined and shown. Each of them showed 20- to 500-fold greater yields than an irrelevant bacteriophage. First round, uPAR-expressing S59 cells were used; Sf9 insect cells expressing the human substance P receptor were used to remove nonspecifically adherent bacteriophage. Second round, Eluted, amplified bacteriophage were affinity selected on transfected COS-7 cells overexpressing human uPAR. Third round, affinity selection was done on uPAR-expressing S59 cells again. 977 CSWRPPFRAVC(4)[4.2 ± 0.7] CSWAPPFKASC(3)[16.1 ± 6.8] CNWTPPFKTRC(2)[1.6 ± 0.5] W-x-P(2)-F-[KR] 3 Phage display (common panning) 3 PMID:8352728 Biotin Streptavidin 3MG5,1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F, Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA Binding of biotinylated proteins to immobilized phages was determined by ELISA. Absorbance at 405 nm was measured. Data are means ± S.E.M. from 2-4 experiments and expressed as the ratio of absorbance readings, obtained in the presence of test protein to that measured in its absence. Three rounds of biopanning was performed using biotinylated mAb 35 (a biotinamidohexanoate conjugate of the anti-acetylcholine receptor monoclonal antibody) at concentrations of 1000, 10 and 1 nM respectively. The inability of B-mAb 35-binding phages to recognize the non-biotinylated antibody was confirmed by competition experiments. 978 CLARSRLPAIPS(9) SRMSPLVPLRNS(1) 2 Phage display (common panning) 4 PMID:8596952 Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. [SC]X10[SC] phage display library Residues 81 to 140 of chicken c-Src were cloned into the Hind III-Bam HI sites of the plasmid pMMHb and expressed in Escherichia coli. This SH3 domain of chicken c-Src was used as the target. 979 CWYLGYWPGQEC(12) CKRFVWRGQALC(10) CLMGLRLGLLPC(4) CLSGLRLGLVPC(2) CAYGFKLGLIKC(1) 5 Phage display (common panning) 4 PMID:8596952 Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. [SC]X10[SC] phage display library The D-amino acid SH3 domain of chicken c-Src (residues 81 to 140) were prepared by chemical synthesis. D-SH3 was used as the target. After four rounds of selection, 29 clones were sequenced, of which only 7 clones bound D-SH3. To ensure that the selected phages were not binding to streptavidin or to a composite surface formed by the streptavidin-D-SH3 complex, a fifth selection round was performed with neutravidin as a matrix. Sequence analysis of clones after this fifth round of selection revealed only sequences CLMGLRLGLLPC(12). Its D-peptide bound to L-SH3 domain of chicken c-Src. The D-peptide of CLSGLRLGLVPC also bound to L-SH3. CKRFVWRGQALC resembled the background sequences that were isolated with a variety of biotinylated ligates. Indeed, its D-amino acid version failed to bind to the L-SH3 domain, as judged by ELISA and NMR studies. CWYLGYWPGQEC showed limited similarity to the first sequence and has not been examined further. 980 CLSSRLDAC CNSRLHLRC CENWWGDVC CKDWGRIC CVLRGGRC CTRITESC 6 Phage display (in vivo) 3 PMID:8598934 Brain blood vessels Not determined. Not determined. Not determined. CX5C+CX6C+CX7C+CX9 phage display library pool Forty-eight brain-localizing phage were sequenced. Peptides containing an SRL motif represented 54% of the clones, followed by CENWWGDVC. Other clones that appeared more than onece included CKDWGRRIC,CVLRGGRC and CTRITESC. 981 WRCVLREGPAGGCAWFNRHRL(10) 1 Phage display (in vivo) 3 PMID:8598934 Brain blood vessels Not determined. Not determined. Not determined. X2CX14CX2+X2CX18 phage display library pool Twenty-five brain-localizing phage were sequenced. The WRCVLREGPAGGCAWFNRHRL phage targeted the brain 9-fold more effectively than the kidney. 982 CKGRSSA(3) CLPVASC(1) CGAREMC(1) 3 Phage display (in vivo) 3 PMID:8598934 Kidney blood vessels Not determined. Not determined. Not determined. CX5C+CX6C+CX7C+CX9 phage display library pool Forty-eight kidney-localizing phage were sequenced. Peptides CLPVASC and CGAREMC represented 60% of the clones. The CLPVASC phage targeted the kidney 7-fold more effectively than the brain. 983 AAVERSKMIDRNLRE(3) AALERSKAIEKNLKE(1) EAARRSRRIDRHLRS(1) HSRELEKKLKE(1) ELAKRSKELEKKLQE(1) AQREANKKIEKQLQK(1) E-R-x(2)-A-x(3)-N-L 6 Phage display (common panning) 3 PMID:8654559 Anti-G(o) monoclonal antibody 3E7 Guanine nucleotide-binding protein G(o) subunit alpha Not determined. Not determined. f3-15mer phage display library (X15) 984 CRIGPITWVC[10] 1 Phage display (common panning) PMID:8662529 Erythropoietin receptor Erythropoietin 1CN4,1EER, 1EBA,1EBP, ON1203 phage display library (CX8C) Binding assay Concentrations required for 50 percent inhibition of tracer (125)I-labeled EPO binding (IC50 values, μM) were determined in a competition binding assay. Under the assay conditions, the IC50 was approximately equal to the affinity (Kd). The extracellular domain of the human EPO receptor (EPOR ECD) was expressed in Chinese hamster ovary cells as a fusion protein. Besides EPOR ECD, this fusion protein had a COOH-terminal signal sequence of the human placental alkaline phosphatase (HPAP) to aid in purification and immobilization. Between the ECD and HPAP domains, a protease (thrombin) cleavage site was inserted. The fussion protein was used as the target; target-phage complexes were cleaved from mAbl79 with bovine thrombin at room temperature for 10 minutes. EPO was found to compete with the clone displaying CRIGPITWVC for binding to the immobilized EPOR. The peptide GGCRIGPITWVCGG was capable of competing with EPO binding to immobilized EPOR with an apparent affinity of approximately 10 uM. Higher affinity peptides were found from a secondary mutagenesis library containing peptides of increased length and were verified as full EPOR agonists in vitro and in vivo. The stuctures of EPO mimetic peptides complexed with EPOR have been solved (e.g. 1EBA, 1EBP). 985 WPLYSTLP(1) SPIYEPIY(1) IVSYLPIY(1) SLIYDALP(1) EPIYGPVP(1) VPIYDTLV(1) EEIYLRWY(1) APIYWTFD(1) FDLYWRLP(1) YSVYEAWG(1) YPIYDYIT(1) IYDYLPWS(1) LYDYLPIF(1) 13 Phage display (common panning) 3 PMID:8709147 Tyrosine-protein kinase Blk Not determined. Not determined. Not determined. X3YX4 phage display library Human Blk was expressed in insect cells by baculovirus and 38 enzyme units of Blk were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 986 IYDYLPWP(8) FHIYDFLP(4) ASIYGCWL(1) APIYEVLR(1) WPIYEPLR(1) WPIYEPLW(1) WPIYEVLP(1) SPLYPFVG(1) SELYSPIF(1) EPIYDVIF(1) EPVYDVLP(1) RDIYDWLP(1) GPIYEPLY(1) DPIYATWG(1) VYDYLPFR(1) VYDYVPFH(1) DYFYDYLI(1) 17 Phage display (common panning) 3 PMID:8709147 Tyrosine-protein kinase Blk Not determined. Not determined. Not determined. X3YX4 phage display library Human Blk was expressed in insect cells by baculovirus and 3.8 enzyme units of Blk were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 987 IYDYLPWS(5) FPIYDFLP(2) EPVYDVLP(1) EDFYDFLP(1) DSIYDELH(1) ISIYDFLP(1) YPIYDTLP(1) DDIYWYLP(1) VYDYLPFP(1) 9 Phage display (common panning) 4 PMID:8709147 Tyrosine-protein kinase Blk Not determined. Not determined. Not determined. X3YX4 phage display library Human Blk was expressed in insect cells by baculovirus and 3.8 enzyme units of Blk were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 988 DVLYAPLR(1) DDAYYWFF(1) DSIYDWIF(1) ESLYWSWP(1) EELYGTLG(1) EPIYDSLP(1) EPVYVIFR(1) ESLYSSFH(1) WLIYEYLP(1) YEIYEYPP(1) LYDYLPIR(1) IYEYLPRF(1) 12 Phage display (common panning) 3 PMID:8709147 Lyn SH3 domain of tyrosine-protein kinase Lyn Not determined. Not determined. Not determined. X3YX4 phage display library Human Lyn was expressed in insect cells by baculovirus and 60 enzyme units of Lyn were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 989 SPLYDFIP(2) IYDYLPWS(2) DPVYDLLP(1) DGIYEVLP(1) DPLYHWIP(1) DDLYAELP(1) DNIYATLP(1) DDIYWYLP(1) EPIYDVLF(1) EPLYAPIR(1) EDIYWLLP(1) PEIYWSFP(1) PTFYDFLA(1) FPIYDFLP(1) SAIYLPAL(1) SPLYESLP(1) GPIYELLP(1) DYEYPSVL(1) 18 Phage display (common panning) 3 PMID:8709147 Lyn SH3 domain of tyrosine-protein kinase Lyn Not determined. Not determined. Not determined. X3YX4 phage display library Human Lyn was expressed in insect cells by baculovirus and 6 enzyme units of Lyn were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 990 IYDYLPWS(4) DDIYWYLP(3) DPIYEELP(2) WDIYELLP(2) DGIYEVLP(1) EPLYEELP(1) SPVYEFLP(1) YHSYPPPV(1) 8 Phage display (common panning) 4 PMID:8709147 Lyn SH3 domain of tyrosine-protein kinase Lyn Not determined. Not determined. Not determined. X3YX4 phage display library Human Lyn was expressed in insect cells by baculovirus and 6 enzyme units of Lyn were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 991 ESVYGIFH(1) EPLYGTFH(1) EPLYGSCC(1) EDLYGIWA(1) EFTYWMFG(1) EDLYWAYP(1) EMLYWDYY(1) ELLYWPWY(1) ELDYWSFS(1) ELLYWSLP(1) EPLYGIFE(1) EPIYSSYE(1) ESLYWSWP(1) ESLYWAYH(1) ESLYWYWP(1) ELLYWSFP(1) DNVYWSFG(1) DPIYWVVH(1) DQLYWSFG(1) YWGYGDEW(1) YWTYSPPM(1) 21 Phage display (common panning) 3 PMID:8709147 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. X3YX4 phage display library The catalytic domain of human c-Src was expressed in E. coli. and 35 enzyme units of Src were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 992 DDLYGEFH(1) QPIYSDFY(1) QLIYWDCC(1) EELYWSFF(1) ESIYGIFH(1) DLIYGEWS(1) YWIYPWID(1) 7 Phage display (common panning) 3 PMID:8709147 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. X3YX4 phage display library The catalytic domain of human c-Src was expressed in E. coli. and 3.5 enzyme units of Src were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 993 ESIYWAFE(2) EPIYGTWI(2) ESLYWSWP(2) ENLYGAFP(1) ESLYATFL(1) EPIYWALP(1) EPIYGAVP(1) EPLYWVCC(1) EPIYWSFD(1) EPIYGSWY(1) ESIYGIFH(1) DNLYWSFP(1) DDLYWSWS(1) DLLYWDFY(1) 14 Phage display (common panning) 4 PMID:8709147 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. X3YX4 phage display library The catalytic domain of human c-Src was expressed in E. coli. and 3.5 enzyme units of Src were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 994 WPDYESFH(1) LSDYEGPF(1) DSDYEWPS(1) DEDYDYAL(1) FSDYEYPT(1) EPEYWFPS(1) YEDYEQWP(1) YPDYELPP(1) YSDYEAPW(1) WPDYEPAF(1) DYEYLSPW(1) DYEYLSPL(1) 12 Phage display (common panning) 3 PMID:8709147 Tyrosine-protein kinase SYK Not determined. Not determined. Not determined. X3YX4 phage display library Human Syk was expressed in insect cells by baculovirus and 16 enzyme units of Syk were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 995 YEDYEQWP(3) TDDYEFPV(2) WLIYEVLR(1) LPDYEVLH(1) DQDYWYPW(1) LPDYELIP(1) DPDYEWPI(1) EPDYIYFP(1) DSDYEWPS(1) TEDYEGPS(1) PPDYEDVY(1) MSDYEYPM(1) DSPYEWWP(1) EDDYSWPF(1) DPIYVSFT(1) 15 Phage display (common panning) 3 PMID:8709147 Tyrosine-protein kinase SYK Not determined. Not determined. Not determined. X3YX4 phage display library Human Syk was expressed in insect cells by baculovirus and 1.6 enzyme units of Syk were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 996 LPDYEFPS(1) DDDYIMLW(1) QDLYGSWG(1) DDIYWYLP(1) IGDYEFPA(1) ADDYEYPF(1) EDLYEWPY(1) DSDYEWPS(1) EPDYWWMP(3) YPDYEISS(1) YEDYEHWP(1) YEDYEQWP(5) 12 Phage display (common panning) 4 PMID:8709147 Tyrosine-protein kinase SYK Not determined. Not determined. Not determined. X3YX4 phage display library Human Syk was expressed in insect cells by baculovirus and 1.6 enzyme units of Syk were used to phosphorylated the phage library. The phosphorylated library was subjected to affinity selection on the anti-phosphotyrosine monoclonal antibody (4G10) column. 997 DRSYLSFIHLYPELA(8)[0.68, 0.15] SARLWAEYLPLYRHM(2)[0.78, 0.14] FNGGAQMGWDYYWFF(1)[0.23, 0.15] WDAMYWNWRSVSEFH(1)[0.19, 0.08] 4 Phage display (common panning) 3 PMID:8756700 Toxic shock syndrome toxin-1, TSST-1 MHC class II molecules Not determined. Not determined. f3-15mer phage display library (X15) ELISA Absorbances at 450 nm were measured in a noncompetitive ELISA and a competitive ELISA, respectively. In a competitive ELISA, 1 μg of native TSST-1 was added to each well to compare with the immobilized TSST-1 for binding to individual phage. Data shown were reproduced from the graph. Selected phage were found to react specifically with TSST-1 but not with other staphylococcal exotoxins. The synthetic peptide (DRSYLSFIHLYPELA) was shown to inhibit binding of all four isolated phage to TSST-1, suggesting that they bind to a common site on TSST-1. Furthermore, DRSYLSFIHLYPELA was shown to compete with MHC class II molecules (DR Antigens purified from the human B lymphoblast cell line Daudi) for binding to TSST-1 in a concentration-dependent manner. 998 RWFHRH(2) YLRWMY(1) PMWMIG(1) RWSDMA(1) MSYLEG(1) LIYLVN(1) YGYLVN(1) TRVPRR(2) SRLPLR(4) SRLPKR(1) TGTLLL(1) VWSVFL(1) FGRRAE(1) SGSSTP(1) FVYHLL(1) SRHRHH(2) CIPTFF(1) VSRLIV(1) RLLWRV(1) RSSMSI(1) DYRSCL(1) 21 Phage display (common panning) 3 PMID:8783147 Anti-HBsAg polyclonal antibody IgG Hepatitis B surface antigen Not determined. Not determined. f3-6mer phage display library (X6) The binding of phage displaying STGPCR and TRVPRR to immune serum IgG was about four times better than to pre-immune serum IgG. 999 AYSKTPPPIP(3) PYLKTPPPIP(1) FHFRPPPPIP(1) GYNKPPPPIP(1) PYGKVPPPIP(1) AYGKPPPPIP(1) 6 Phage display (common panning) 4 PMID:8810341 Tyrosine-protein kinase ITK/TSK Not determined. Not determined. Not determined. X6PPIP phage display library The Itk SH3 domain was amplified from the murine cDNA clone by polymerase chain reaction. The product, encoding amino acids 171 to 232, was cloned into the BamHI site of pGEX-2T (Pharmacia). The W208K mutation was generated by polymerase chain reaction and subcloned into this construct. GST-Itk SH3 fusion protein was produced in Escherichia coli, purified and immobilized on polystyrene as the target. 1000 DRSSWSKPPPIP(2) SKMGWSKPPPIP(1) MGSGWSKPPPIP(1) LKEGWSKPPPIP(1) VNQGWSKPPPIP(1) MNNSWSKPPPIP(1) YSSMWSKPPPIP(1) GNNSWSKPPPIP(1) HHYGWSKPPPIP(1) FGRSVSKPPPIP(1) VANGHSKPPPIP(1) VYRGWSKPPPIP(1) RGLGWSKPPPIP(1) VSSTWSKPPPIP(1) SKNNWSKPPPIP(1) VNSKWSKPPPIP(1) AHSGWSKPPPIP(1) LREGWSKPPPIP(1) FKSGWSKPPPIP(1) WNDTWSKPPPIP(1) GVSNWSKPPPIP(1) CNEGWSKPPPIP(1) 22 Phage display (common panning) PMID:8810341 Tyrosine-protein kinase ITK/TSK Not determined. Not determined. Not determined. X5YSKPPPIP M13 phage display library The Itk SH3 domain was amplified from the murine cDNA clone by polymerase chain reaction. The product, encoding amino acids 171 to 232, was cloned into the BamHI site of pGEX-2T (Pharmacia). The W208K mutation was generated by polymerase chain reaction and subcloned into this construct. GST-Itk SH3 fusion protein was produced in Escherichia coli, purified and immobilized on polystyrene as the target. 1001 DKMGRTSRW(2) WAGRSISYK(2) WTVRTLSSR(2) WYTAQRSLY(1) WTSVMVSAF(1) FWYWHTTQR(1) FWEVVTITN(1) LISWENFAQ(1) WEIKSLSGI(1) WVVVTLNLW(1) WMAMSVSEK(1) QVGPYEWRR(1) SRHWEGRGK(1) WGSRAESRV(1) YGPGLRWKS(1) SWTVVHISS(1) WVLRSIRAQ(1) ATWAEQWRT(1) WGEDPEARR(1) WKMSSWRWD(1) YKMGRTSRW(1) GYKPRWHNC(1) IGEQGWRHR(1) AQVWGRRRM(1) WAGPTPSDE(1) LQHRWHKLE(1) VRALWEAKS(1) FMTMWAWET(1) STRGWPWAR(1) WPIVDVSAP(1) GWAIESIRW(1) AVLWWFNER(1) LEVPLRQHY(1) RRGEVLGAR(1) NLGRMGARH(1) EIRQALARE(1) DRNLEGRGK(1) ELRRFLELL(1) EALEQLGAL(1) LRSTNARSA(1) VITPDPSEV(1) RSQIPVREL(1) QIRMNTNEF(1) FPARSISHK(1) FIRKHLNKL(1) GIGMGHSEQ(1) MKAVIGRTM(1) GRQGRGDMW(1) EARGEVWGR(1) CARSGMSGW(1) NEMTTMIEW(1) GQQERVWPS(1) PDSQQINLE(1) NTHKDRVHL(1) HQKRVEESA(1) 55 Phage display (common panning) 3-4 PMID:8881746 HLA class II histocompatibility antigen DR13 Not determined. Not determined. Not determined. M13lib X9 phage display library HLA-DR13 antigen was purified from the Epstein-Barr virus-transformed B-cell line HHKB (DRB1*1301, DRB3*0101) and used as the target. HLA-DRB1*1301 and *1302 alleles differ only at position 86 of the HLA-DR beta chain, where they contain valine and glycine residues respectively. 1002 YWWTWSRAG(1) WTSRTLSAR(1) WARHRTGSE(1) SVWLRWRGC(1) AAKWQKRVE(1) QAGWYWWVR(1) IGEQRWRHR(1) VVIDRWEIR(1) LGGVHYWRR(1) KGWLGGRAS(1) WRSLRTLLE(1) RKEPWGEMS(1) MEFRAGSHA(1) KSVAFMKGR(1) RSFSRVLEE(1) PMFQLWEGQ(1) TQLRGRRLN(1) AVDNQLHER(1) LHARQLPRG(1) IKSLLRKEL(1) LISPEQPPQ(1) PMSQYSVGQ(1) METYLRISS(1) AIYSRRVLR(1) SYVSYNEGE(1) PISSTRIEG(1) ARQPRGHMW(1) HTRRKWGGE(1) RGGVHDWAR(1) DRRPHNWGE(1) GAPPRAFAG(1) MAQERFLEM(1) SHGPKNFGE(1) AAGASAAQL(1) AGRACALAG(1) PSATSPLEK(1) TQGKRGYNS(1) RQTVEGHRL(1) PVNDHPLEK(1) SMAATVGAR(1) GGAGVAKHE(1) KSGKEVERR(1) SDTCTCHTR(1) TSRHEQARN(1) SRGGSNMRE(1) RSTSSCSER(1) 46 Phage display (common panning) 3-4 PMID:8881746 HLA class II histocompatibility antigen DR13 Not determined. Not determined. Not determined. M13lib X9 phage display library HLA-DR13 antigen was purified from the Epstein-Barr virus-transformed B-cell line WT-47 (DRB1*1302, DRB3*0301) and used as the target. HLA-DRB1*1301 and *1302 alleles differ only at position 86 of the HLA-DR beta chain, where they contain valine and glycine residues respectively. 1003 FGRIPSPLAYTYSFR(15) HRWMPHVFAVRQGAS(12) VSWFSRHRYSPFAVS(11) QLQSYRFFFPSYMGG(5) WSNRMPPLFTPWYPP(3) YWALHHSGWPFSRGS(2) GYPWRIRPWASGPFL(2) RRLVFWHGFETTGPR(2) DRWRPALPVVLFPLH(2) GPWYCTLGLCHFRSS(2) GWPSFSNHPFLYPRW(2) LNPFRSLFFPALDNL(2) LSWPLHAGRGFRVWS(2) VGFLGLKRGPPGVDA(2) GRWAFAPSSWHLYSR(2) GAGMLRWFGYPALYG(2) SRVRFPAWGLPFSPV(1) WFPGPITFIPRPWSS(1) WWMSRPSRLLYYEYG(1) HRVQFAGWGFPGFRL(1) WHWRLPRSTWHPTSV(1) 21 Phage display (common panning) 7 PMID:8889832 Integrin alpha-6-beta-1, integrin α6β1 Laminin-1 Not determined. Not determined. f3-15mer phage display library (X15) The integrin alpha-6 beta-1 was isolated from human placenta and used as the target. The synthetic peptide VSWFSRHRYSPFAVS was found to strongly inhibit the binding of laminin-1 to integrin alpha-6 beta-1. This inhibitory effect seemed to be specific for alpha-6 beta-1 integrin, since it did not affect the binding of fibronectin to integrin alpha-5 beta-1. 1004 EWCEYLGGYLRCYA(38) 1 Phage display (competitive panning) 3 PMID:8953648 Intercellular adhesion molecule 1, ICAM-1 Integrin alpha-L beta-2 1MQ8,3TCX, Not determined. X2CX4-8CX2 phage display library pool Phage were incubated with immobilized ICAM-1(1-453) and then eluted sequentially with soluble ICAM-1(1-453), M174F5B7 (a neutralizing anti-ICAM-1 monoclonal antibody) and then with 0.1 N glycine-HC1, pH 2 in the first round. In the second and third rounds of biopanning, a single elution step was performed with the correspongding reagent. Sequencing results revealed that identical phage (EWCEYLGGYLRCYA) were isolated by elution with soluble ICAM-1 (14/14), with antibody (12/12) and with acid (12/12). Phage displaying EWCEYLGGYLRCYA did not bind BSA or other proteins such as streptavidin, but bound to immobilized ICAM-1(1-453) and to ICAM-1(1-185), which contains only the two amino-terminal immunoglobulin domains residing within residues 1-185. This is the region of the ICAM-1 that is bound by LFA-1. The phage did not bind to proteins other than ICAM-1. The linear form of the synthetic EWCEYLGGYLRCYA peptide was found to inhibit LFA-1 binding to immobilized ICAM-1(1-453) in a protein-protein binding assay. By contrast, the disulfide, cyclized, form of the peptide was inactive. 1005 MHCEWFGGPGCVQ GVCDANDGPGCNM DVCRDGGPGCPS DVCRMGGPGCGQ DVCGRGGPGCNS DVCMIGGPGCPG DVCGTGGPGCPV DVCKVGGPGCLP AVCNNGGPGCPS NVCSYQGPGCPG DVCWYAGPGCGI KVCQVWGPGCPI GTCTEDHRGYCHA QACHRGLDHSACFA D-V-C-x(2)-G(2)-P-G-C, H-x-G-[AY]-x-H 14 Phage display (competitive panning) 3 PMID:9237196 Anti-CD80 monoclonal antibody L307.4 T-lymphocyte activation antigen CD80 Not determined. Not determined. X2CX4-8CX2 phage display library pool In each round of biopanning, bound phage were eluted using either nonspecific low pH or soluble human B7-1/Fc fusion protein. The eluted phage bound to L307.4 but not to an isotype matched antibody, indicating that binding was antibody specific. Synthetic peptides inhibited phage binding to L307.4, indicating that the gene III protein is not required for peptide binding. In addition, the cyclized forms of synthetic peptides containing the DVCXXGGPGC motif were capable of inhibiting L307.4 binding to soluble B7-1/Fc fusion. Phage expressing only the HXG(A/Y)XH consensus sequence were inhibited from binding to L307.4 by the presence of chelating agents. 1006 TPTTIIHHYGAQHFDMKDST ADPSMHHHLGAVHARTGLTV QYPDWHHHRGAVHAQSGPNR HVTHNRPPAGAHVGYQHEER VGSNSRHHLGALHDPEMWER SLPHTGYHHGASGHPLFMDW TTWQLQHHLGAQHEYVAGWW HKGYIHIGPGAWQTVASERL QTDRTPHHLGAFHENQLESY HLGYLHLPFGAERTGTGDVH ILGPLSHHQGAWHYGAPVTE NPTPRDHHFGALHYPVEPGW ARATNLHHMGARHDSPFVAL QSNYPMHHLGAWHEQFELSF TPMPVGHHKGAQHVVQPGDT GPPGRGDTTGAHLGYSHAMP HMGWRHFPTGAYEDHGPSEM H-x-G-[AY]-x-H 17 Phage display (competitive panning) 3 PMID:9237196 Anti-CD80 monoclonal antibody L307.4 T-lymphocyte activation antigen CD80 Not determined. Not determined. X10+X9GAX9 phage display library pool In each round of biopanning, bound phage were eluted using either nonspecific low pH or soluble human B7-1/Fc fusion protein. The eluted phage bound to L307.4 but not to an isotype matched antibody, indicating that binding was antibody specific. Synthetic peptides inhibited phage binding to L307.4, indicating that the gene III protein is not required for peptide binding. Phage expressing only the HXG(A/Y)XH consensus sequence were inhibited from binding to L307.4 by the presence of chelating agents. 1007 YMRYYESSLKSYPDW(16) TMTFPENYYSERPYH(2) PPPIFRYYEYWPTSY(1) 3 Phage display (common panning) 3 PMID:8954559 Alpha-bungarotoxin Acetylcholine receptor subunit alpha 1IDG,1IDH, Not determined. X15 M13 phage display library HPLC-purified alpha-bungarotoxin from the snake venom of Bungarus multicinctus was biotinylated and attached to an immobilized nitrostreptavidin matrix. Nitrostreptavidin reversible biotin-binding properties was prepared by chemical modification of the protein using tetranitromethane. Bound phage were eluted with 0.1M HCl titrated to pH 2.2 with glycine. 1008 IWRYYEDSELMQPYR(10) EYMRYYESSLNPTRL(3) YMRYYESSLKSYPDW(1) 3 Phage display (competitive panning) 3 PMID:8954559 Alpha-bungarotoxin Acetylcholine receptor subunit alpha 1IDG,1IDH, Not determined. X15 M13 phage display library HPLC-purified alpha-bungarotoxin from the snake venom of Bungarus multicinctus was biotinylated and attached to an immobilized nitrostreptavidin matrix. Nitrostreptavidin reversible biotin-binding properties was prepared by chemical modification of the protein using tetranitromethane. Bound phage were eluted with 0.1 mg/ml biotin. 1009 GRVCLR(2)[92, 30] YSPELE(1)[120, 13] YSPEVR(1)[62, 11] YSPEVG(1)[75, 26] YSTEVR(1)[67, 20] YSPHLR(1)[54, 12] GPVLRR(1)[92, 25] 7 Phage display (common panning) 3 PMID:8960114 Anti-MSP1 monoclonal antibody D14-3 Major blood-stage surface antigen Pv200 Not determined. Not determined. CX6C phage display library ELISA measurements of the binding to Dl4-3, expressed as the percentage of the signal given by pC3H-Pv42 (phagemid bearing the whole pV42 protein). ELISA measurements of the competition with Pv42, expressed as the inhibition percentage of the binding of Pv42 to D14-3. These phagotopes were injected into mice belonging to Balb/c, lC57B1/6 and Biozzi strains. All animals developed a strong immune response against phage particles but only Biozzi mice produced antibodies cross-reacting with the Mr 42000 C-terminal fragment (Pv42) of Plasmodium vivax merozoite surface protein 1 (PvMSPl). All phagotopes in Biozzi mice elicited a specific response against Pv42, even those sharing no sequence similarity with the antigen. 1010 WDTVRISF WTPSASRF WESVRTHF WSPSASRF WASVRTHF WDTVRICF WESCGTHF WPSLQAIR WPTLSKIA WPRPCRHS WPPPARVI WPQLQRLI WPPLSSVL 13 Phage display (common panning) 3 PMID:9003408 Calmodulin, CaM Not determined. Not determined. Not determined. X8 phage display library WDTVRISF is the most frequent sequence, represented 60% of all sequences obtained. WPSLQAIR is the second most frequent sequence, represented 20% of all sequences obtained. NMR spectroscopy and fluorimetry indicateed that both peptides interact with CaM in the presence of Ca2+. The two peptides differentially inhibited CaM-dependent kinases I and II (CaM kinases I and II) but did not affect CaM-dependent phosphodiesterase. WDTVRISF inhibited CaM kinase I but not CaM kinase II, whereas WPSLQAIR inhibited CaM kinase II, but only partially inhibited CaM kinase I at a more than 10-fold higher concentration. 1011 HPWGYPKI HRGGTPKI HPWTRTIS HPWTRTDS WNITWSFS FQNMRMAG PAKPRAAT FHNQRMAG GTHYHASP 9 Phage display (common panning) 3 PMID:9003408 Aequorin Not determined. Not determined. Not determined. X8 phage display library HPWGYPKI is the most frequent aequorin-binding peptide. 1012 TDVRQGLGWREGVVGLWDRH(1)[560] NRDYDSRSDIWSIWSLGRER(1)[370] VSSPFDFSAHSPIEGLWAGE(1)[40] VSMKLEDKPRTALFDLWQAT(1)[160] TGKDSVVWLWGVN(1)[500] IVSLWD(1)[710] VQERVEVYGLWGGIGNLSAD(1)[NT] HDADAKEGPKRRSASELWGP(1)[NT] ELEENMFCSWKLWGYSCRGP(1)[NT] RGDVPAASARGSVSIRDLWR(1)[NT] YKHGTVRMLWPGGGVRVADG(1)[NT] ESMDRRGVGELWGTPSKSTR(1)[NT] VLNRDGGGSTVEALWGLGYN(1)[NT] NIWIYGVRDLWGPFEPGIVG(1)[NT] EGRSEVDIDDQNTSVYKLW(1)[NT] DTHERAWHLWQGTVSLTRP(1)[NT] RSSILERIWGG(1)[NT] YPRETAIQLW(1)[NT] EEHKLMSVLW(1)[NT] 19 Phage display (common panning) 3 PMID:9039957 Anti-gp120 monoclonal antibody GV1A8 Surface protein gp120, SU Not determined. Not determined. X20 phage display library Western blot,Binding assay Equal amounts of the five phages were analyzed by western blotting using the GV1A8 mAb as the probe. It illustrated that the phage's affinity for GV1A8 could be ranked as Φ35 (VSSPFDFSAHSPIEGLWAGE)> 53 (VSMKLEDKPRTALFDLWQAT) > 30 (NRDYDSRSDIWSIWSLGRER) > 37 (TDVRQGLGWREGVVGLWDRH). This was further confirmed by performing quantitative binding assays in which radioiodinated GV1A8 was used for each of the five selected phages. The Kd (nM) value was shown. 1013 GGETRSNFEAWKWDEAQQGG(1) ILRARENQTRWYRDMGGDMS(1) GFKVEANYNAWREWVRSDWV(1) TNHPGVDADYWRGWKERYNR(1) YWKDWLHDTAGGGVWSRDNH(1) WREYVYLWWIDHKT(1) YLQWVRDTWV(1) 7 Phage display (common panning) 3 PMID:9039957 Anti-gp120 monoclonal antibody GV4D3 Surface protein gp120, SU Not determined. Not determined. X20 phage display library 1014 GRWGGVQKFG KRWFLIGDRM RIDRWGVPFE AEFSFSVTKW DRYDMRNWMA ALGRLKHTSR LSLEKTDNSV FMNSGETRLG 8 Phage display (common panning) 2 PMID:9119446 Anti-GXM monoclonal antibody 2E9 Capsular glucuronoxylomannan (GXM) Not determined. Not determined. X10 fUSE5 phage display library An enzyme-linked immunosorbent assay (ELISA)-based screening method led to the selection of phages with peptide inserts that bound 2E9 and inhibited 2E9-GXM binding. 1015 GIGFQTGLRF KRWFLIGDRM LRWDNFTSKD PLKYWIQVNM NDLESLDRTE 5 Phage display (common panning) 3 PMID:9119446 Anti-GXM monoclonal antibody 2E9 Capsular glucuronoxylomannan (GXM) Not determined. Not determined. X10 fUSE5 phage display library An enzyme-linked immunosorbent assay (ELISA)-based screening method led to the selection of phages with peptide inserts that bound 2E9 and inhibited 2E9-GXM binding. 1016 GIGFQTGLRF(10) PESWHTRWDE(4) PPRVPPDLMS(3) RAYQTTLLTL(2) GMDGTQLDRW(1) TRGFSTDRNS(1) LWNKRTAEFK(1) 7 Phage display (common panning) 3 PMID:9119446 Anti-GXM monoclonal antibody 2E9 Capsular glucuronoxylomannan (GXM) Not determined. Not determined. X10 fUSE5 phage display library An enzyme-linked immunosorbent assay (ELISA)-based screening method led to the selection of phages with peptide inserts that bound 2E9 and inhibited 2E9-GXM binding. The synthesized peptide GMDGTQLDRW inhibited GXM binding to solid-phase 2E9 and 2E9 binding to solid-phase GXM. It also inhibited the GXM binding of GXM-TT immune sera. 1017 RHMREESFSRPFVVA E(2)-x-F 1 Phage display (common panning) 4 PMID:9128182 Anti-dystrophin monoclonal antibody MANDYS142 Dystrophin Not determined. Not determined. f3-15mer phage display library (X15) Only one sequence was given in the original paper. Of the 24 randomly selected clones, 23 clones were recognised by both MANDYS141 and MANDYS142. The left one clone failed to react with either MANDYS141 or MANDYS142. 1018 LPNSCTANTYWCNDME(2)[58.58 ± 5.87] STSLCTMSTYYCADTE(2)[60.42 ± 4.83] AANDCTHNTYWCTYYG(1)[76.80 ± 5.71] IRLDCAENHQKCQLME(1)[NT] DQIDCTKELQLCQLKE(1)[NT] ASDRCRQDTFFCDWRI(1)[NT] APKTCAHSTYYCSYEM(1)[NT] SNNNCTNQVAWCQMQV(1)[NT] 8 Phage display (common panning) 3 PMID:9174614 Anti-DNA monoclonal antibody F14.6 Deoxyribonucleic acid, DNA Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) Surface plasmon resonance (SPR) Inhibition of binding of 3.75 μg/ml mAb F14.6 on FC2 oligonucleotide immobilized on a sensor chip by the addition of 3e12 phages/ml. Binding of free mAb is monitored by surface plasmon resonance (BIAcore). Results are presented as the percent inhibition and calculated as follows: (RU control - RU phage/RU control) x 100. Data shown were reproduced from the graph. NT denotes not tested. Binding of all phages selected on F14.6 was inhibited with 700 ng/ml soluble DNA. BALB/c mice immunized with LPNSCTANTYWCNDME-bearing phage exhibited high serum titers of IgG3 anti-dsDNA antibodies. 1019 DMMRCTTTRWKCGTDM(1) AKYSCQKTMCYGLV(5) 2 Phage display (common panning) 1 PMID:9174614 Anti-DNA monoclonal antibody J20.8 Deoxyribonucleic acid, DNA Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) The binding of AKYSCQKTMCYGLV phage to J20.8 could not be inhibited with soluble DNA. 1020 MKKACLERAINCVPTL(1) GNSNCWIWGLHCTPRS(2) LLGDCSQWGPECKVRR(3) VTGDCKYWPGQCEAHF(2) LTGDCGYEGVTCRLGK(8) VVGDCMSTWAFCATIW(1) VMGDCNESNAFCAHLR(1) RPQNCNLTTQPCDEIP(1) GNSNCAKDHPTCEWHQ(1) GSRNCPTTKTHCRDTT(1) 10 Phage display (common panning) 2 PMID:9174614 Anti-DNA monoclonal antibody J20.8 Deoxyribonucleic acid, DNA Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) 1021 LTGDCGYEGVTCRLGK(8)[52.16 ± 6.18] AKYSCQKTMCYGLV(5)[NT] LLGDCSQWGPECKVRR(3)[NT] VVGDCSAWQGACQSHS(2)[81.58 ± 16.88] VTGDCKYWPGQCEAHF(2)[NT] IKGDCLGWQVQCMAAV(2)[NT] GNSNCWIWGLHCTPRS(2)[NT] ITGDCTSEVWQCYGGV(1)[49.62 ± 5.39] MTGDCKHWQNACFWSS(1)[67.76 ± 6.01] LTGDCQKWXMSCFINK(1)[NT] LSGDCWNWIQGCRLPF(1)[NT] LIGDCNNWLERCNIPR(1)[NT] VQGDCWGWDNDCGSRN(1)[NT] PAWDCWAWRWQCNSYD(1)[NT] RTHTCYRKHSWCTT(1)[NT] 15 Phage display (common panning) 3 PMID:9174614 Anti-DNA monoclonal antibody J20.8 Deoxyribonucleic acid, DNA Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) Surface plasmon resonance (SPR) Inhibition of binding of 3.75 μg/ml mAb J20.8 on FC2 oligonucleotide immobilized on a sensor chip by the addition of 3e12 phages/ml. Binding of free mAb is monitored by surface plasmon resonance (BIAcore). Results are presented as the percent inhibition calculated as follows: (RU control - RU phage/RU control) x 100. Data shown were reproduced from the graph. NT denotes not tested. Binding of all phages selected on J20.8 were inhibited with soluble DNA except phage displaying AKYSCQKTMCYGLV. BALB/c mice immunized with phage dsiplaying LTGDCGYEGVTCRLGK or ITGDCTSEVWQCYGGV exhibited high serum titers of IgG3 anti-dsDNA antibodies. The peptides selected on J20.8 also bound serum antibodies from human patients with systemic lupus erythematosus. 1022 GAVITH(1) RDIVVA(1) VYSHAS(1) HSYSAG(1) LDIVVA(1) 5 Phage display (common panning) 7 PMID:9174952 Chymotrypsinogen A Not determined. Not determined. Not determined. X6 phage display library The five peptides were chemically synthesised and they could competitively inhibit binding of the peptide phage clone to alpha-chymotrypsin. GAVITH, VYSHAS and HSYSAG did not have the ability to inhibit the enzymic activity of alpha-chymotrypsin. RDIVVA and LDIVVA could not be investigated due to their strong hydrophobicity. 1023 YMRYYESSLKSYPDW(16) FTYYQSSLEPLSPFY(3) TMTFPENYYSERPYH(2) PPPIFRYYEYWPTSY(1) HDKLFTFYQNSXSSY(1) Y(2)-x-S(2)-L 5 Phage display (common panning) 3 PMID:9177167 Alpha-bungarotoxin Acetylcholine receptor subunit alpha 1IDG,1IDH, Not determined. X15 M13 phage display library The library-derived peptide (MRYYESSLKSYPD) bears amino acid sequence similarities to a region of the alpha-subunit of the Torpedo muscle AcChoR, as well as of other muscle and neuronal AcChoRs that bind alpha-BTX. 1024 QRGRKA(1)[430] HYGRSG(1)[322] ERARGA(1)[52] ALRRGD(1)[NT] DYRGRM(1)[NT] ERLRKA(1)[NT] FGRHAA(1)[NT] FLPRTA(1)[NT] FRGRAA(1)[NT] HRMRMG(1)[NT] IMRRGK(1)[NT] ITYGRR(1)[NT] KFTRSG(1)[NT] LIPRRA(1)[NT] MTRKRM(1)[NT] NFARMG(1)[NT] NHLRKA(1)[NT] NVGRMG(1)[NT] NVSRRG(1)[NT] PISRRA(1)[NT] PVGRMG(1)[NT] RLLRSV(1)[NT] SFGRRH(1)[NT] SLRGRS(1)[NT] TVLRRA(1)[NT] VARRVK(1)[NT] VIARSN(1)[NT] VNTKSG(1)[NT] VRARGA(1)[NT] VRRGRS(1)[NT] VRRRGA(1)[NT] TRVRAK(1)[NT] 32 Phage display (subtractive panning) 3 PMID:9195973 Tissue-type plasminogen activator Not determined. Not determined. Not determined. X6 fAFF1-tether C phage display library Dot blot,Binding assay As the results of the dot blot assay, phage 7 containing the hexamer FRGRAA was a t-PA-selective substrate. The dot blot assay can rapidly provide information regarding both the activity and specificity of individual substrate phage clones. Based on the results of these assays, three peptide substrates (ERARGA, HYGRSG and QRGRKA) were synthesized and characterized to provide a quantitative analysis of the properties of putative t-PA-selective substrates. These peptides were cleaved 180-1500-fold times more efficiently by t-PA than a control peptide (kcat/Km, 0.29 1/ms) containing the physiological cleavage site present in plasminogen. Kcat/Km (1/ms) were shown. NT denotes not tested. The initial random hexapeptide library fAFF-TC-LIB was subjected to three rounds of high stringency screening with t-PA to prepare an intermediate library containing phage whose randomized hexamer sequences were digested efficiently by t-PA. The intermediate library was then amplified and screened, at low stringency, with u-PA. Following digestion of the intermediate library with u-PA, mAb E-7 and immobilized protein A were added to the mixture, and the resulting ternary complexes were pelleted by centrifugation. The precipitated ternary complexes were retained and the supernatant were discarded, i.e. phage digested by u-PA were substracted. 1025 EHLYMNFP(1) EPPYMNWP(1) EGVYENIP(1) EHVYLNWS(1) EPLYFNWF(1) EPMYQNFS(1) EATYMNWA(1) QPLYMNWM(1) APLYWNWY(1) QHLYMNWM(1) TPVYMNFP(1) NPVYQNWI(1) NPIYQNWI(1) VPIYENFP(1) SHIYENIV(1) SNVYENWT(1) DELYYNWP(1) FRVYENFL(1) PSVYENYT(1) PIIYENYV(1) PGIYWNWF(1) SPIYENFP(1) CLYYNLPY(1) SLYYNWPF(1) Y-[ME]-N-W 24 Phage display (common panning) 4 PMID:9219519 Growth factor receptor-bound protein 2 Not determined. Not determined. Not determined. X3YX4 phage display library Phage were phosphorylated in vitro at an invariant tyrosine residue by a mixture of phosphotyrosine kinases c-Src, Blk and Syk (3500 units, 600 units and 160 units, respectively). Selection of binding motifs was carried out by interaction of the library with the recombinant SH2 domain of Grb2 expressed as a glutathione S-transferase (GST) fusion protein. 1026 TIKGDSRPWTHTRGNVKHYWHAIRSIHMHDRLRSRDFG(1) PALMQEWWNGYAHKELLGPQDATSTHIMHNRIHEEQWD(1) PKLRWQGAAKLTYRFGVNPMYAMLRIYTSFGDHIPWKA(1) GSGGLLGRGGDGVQRISAMYDATHEPVDHSNAIPRAYY(1) ANCKJSDCPLIMWTMWAALHHAMJFRSWGDQFYTHDRS(1) FYRPLASWWHKLMYWFLRIGYARRSIPWQKHGANPMME(1) IEGGLSPRRMSMWEQLHKASDAKQPLLLFGHKANMWPI(1) NRRACMIRRLRIPLSRCHKKYARQLPAFGNBIVYAILH(1) PSRMYDRLHEGFSSWDQSSGNASWSQPTKALEIPGYNN(1) RMSDRLHLSRLPFGGWAQAANANDTPLMMREGSDRQGG(1) 10 Phage display (common panning) 3 PMID:9237207 Anti-PSM monoclonal antibody 7E11-C5 Glutamate carboxypeptidase 2 Not determined. Not determined. D38 phage display library 1027 SGTEGVCTKYSRSGWILTGFGCGASSAPHDREPTSGMHSNLHW(1) GVKGYQSDGGPVPEWPCHANGCGFKPNARKFMHARLRQKNEDW(1) DSQCGVGQCTGFGVCTNGPRGCGIHTLSMTNKLHTPTPPLHAS(1) LRGSASNTYIKHHFRNEAHTGCGKMIRRLRLHKSAGYCGITGS(1) RVFGGETSKGWKSSRMTHVEGCGMSARLHDSSCGSVNYYARPT(1) GYVDPGGVGSCYSMYPGLHRGCGALSHPSIRGPHPGASRRNVH(1) VFGGETSKGWKSMIGYLRSEGCGSRFGLWYCMMSAAIPLVTSN(1) RCSYTNKSAPMYGLLHDGEMGCGNSPSHIIERRIGGSLRPPFR(1) REEARLLMHYRLHKWYSSRAGCGNSNDPCDSRRRGRTHVINRS(1) MMWEQLHPEVCGRAYSTPKRGCGILGDRLHRRSPQYLTPPGR(1) 10 Phage display (common panning) 3 PMID:9237207 Anti-PSM monoclonal antibody 7E11-C5 Glutamate carboxypeptidase 2 Not determined. Not determined. DC43 phage display library 1028 SPEPDWFVEL(30) REPDWYEYVK(11) WHRWPWLVSG(1) WHWWYWALDR(1) EPDW 4 Phage display (common panning) 3 PMID:9237215 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. X10 phage display library Streptavidin-binding peptides containing the consensus amino acid sequence motif EPDW were identified using a phage display library. Phage presenting peptides containing these sequences bound streptavidin in a biotin-sensitive fashion and could be eluted with biotin. Peptides with the WHWWXW motif are nuisance peptides. 1029 CVKWGKKEFC(1) CWQKYWGKEC(1) CYEWGKLRWC(1) CLRWGKWSNC(1) CWRWGKYQIC(1) CVSWGALKLC(1) CIRWGQNTFC(1) CWQWGNLKIC(1) CVRWGQLSIC(1) WGK 9 Phage display (common panning) 3 PMID:9237217 Calmodulin, CaM Not determined. Not determined. Not determined. R8C phage display library Four of the nine peptide sequences (CVKWGKKEFC, CWQKYWGKEC, CLRWGKWSNC and CIRWGQNTFC) were synthesized in cyclized, biotinylated form. All of the peptides required Ca2+ to bind CaM. The cyclized, disulfide-bonded form of CLRWGKWSNC bound CaM better than its reduced form or an analogue in which the cysteine residues were replaced by serine. The cyclized peptide also exhibited the ability to inhibit CaM-dependent kinase activity. 1030 ARVSFWRYSSFAPTY(11) LTLSHPHWVLNHFVS(4) NILSGFHYRNGDGRT(12) HGRFILPWWYAFSPS(38) SLYPGVFALIRSSYP(6) PGSHTPVPRAWAT(5) AREYGTRFSLIGGYR(8) RFRGLISLSQVYLSP(25) VAFVRTYVAGTGGFF(16) GPVWSSGLYRLFYAS(11) NHRLSSPRNPHLYAV(7) GCGRRYDVTCHYVFA(15) TFSGGHKWSISGRSA(11) VSIGFYGRVQYHS(6) 14 Phage display (common panning) 4 PMID:9238628 Thomsen-Friedenreich antigen (T antigen, TF antigen) Not determined. Not determined. Not determined. f3-15mer phage display library (X15) A random 15-amino acid bacteriophage display library was affinity selected against two proteins which display T antigen on their surfaces, asialofetuin and a BSA-T antigen conjugant. Asialofetuin contains three copies of T antigen exposed on its surface, while the BSA-T antigen conjugant has approximately 16 copies of T antigen chemically crosslinked to BSA. Four separate affinity selection procedures were performed with different combinations of antigen, washes and elution conditions. This is a pooled data for the 4 procedures. 1031 YLQASSSS(1) YLQASSSF(2) YLQASSSA(2) YLQASSSL(1) YLQASSSM(1) YLQASASV(1) YLQASAST(1) YLQASASH(1) YLQASTSH(1) YLQASTSF(1) YLQASLSA(1) YLQASLSP(1) YLQASESY(1) YLQASCSL(1) YLQASNSI(1) YLQASSTY(1) YLQASSVF(1) YLQASATY(1) YLQASLTA(1) YLQASMTC(1) YLQASMTQ(1) YLQASTTQ(1) YLQASVVG(1) YLQASAVP(1) YLQASQVF(1) YLQAAERY(1) YLQAHAIY(1) 27 Phage display (common panning) 6 PMID:9325241 Protease precursor, pPR Not determined. Not determined. Not determined. BPTI/Delta-g3p/biased 8-mer phage display library 1032 LVLASQ(1) LVLAST(2) LLCASV(1) GLILAS(1) LLWAST(1) PLVFAS(1) ALLFAS(1) LLMASV(1) LILASG(1) ELLLCS(1) LILASQ(1) LVLASN(1) GLVLAS(1) 13 Phage display (common panning) 6 PMID:9325241 Protease precursor, pPR Not determined. Not determined. Not determined. BPTI/Dg3p/6-mer phage display library 1033 PGYPWH LEYPWH 2 Phage display (common panning) 2-3 PMID:9328567 Anti-Hantaan virus glycoprotein G2 monoclonal antibody mAbGDO5 Glycoprotein G2 Not determined. Not determined. f3-6mer phage display library (X6) Petri dishes were coated with streptavidin and incubated with 5 to 10 mg of biotinylated antibody. Phage-antibody complexes were captured on streptavidin coated petri dishes. 1034 YSDLGK 1 Phage display (common panning) 3-4 PMID:9328567 Anti-P53 monoclonal antibody mAbBp53-11 Cellular tumor antigen p53 Not determined. Not determined. f3-6mer phage display library (X6) Petri dishes were coated with streptavidin and incubated with 5 to 10 mg of biotinylated antibody. Phage-antibody complexes were captured on streptavidin coated petri dishes. 1035 TDAFSDLGRMLANPG 1 Phage display (common panning) 2-3 PMID:9328567 Anti-P53 monoclonal antibody mAbBp53-11 Cellular tumor antigen p53 Not determined. Not determined. f3-15mer phage display library (X15) Petri dishes were coated with streptavidin and incubated with 5 to 10 mg of biotinylated antibody. Phage-antibody complexes were captured on streptavidin coated petri dishes. 1036 RLHYLF RHWSIF RHRTLF RHYLLF MSRHRN RHITLF RHITSL RLHTLF RHFTSL RHHTLL DGARLH 11 Phage display (common panning) 3 PMID:9350871 Phosphoenolpyruvate-protein phosphotransferase Phosphocarrier protein HPr 3EZA,3EZB,3EZE,2XDF, Not determined. f3-6mer phage display library (X6) The peptides were synthesized except RLHTLF, RHFTSL and RHHTLL. All of them inhibited the phosphotransferase system in vitro. 1037 KMSRHRKPGA SSLRGHRWVY KISRHGKRGK KISRHGRPTG RIHFIPRRGR NMSRHRKPTG NMSRHRNPGP NMSRHRIRPT NMSRHRNPTG 9 Phage display (common panning) 3 PMID:9350871 Phosphoenolpyruvate-protein phosphotransferase Phosphocarrier protein HPr 3EZA,3EZB,3EZE,2XDF, Not determined. X10 phage display library The peptides were synthesized except NMSRHRKPTG, NMSRHRNPGP, NMSRHRIRPT and NMSRHRNPTG. All of them inhibited the phosphotransferase system in vitro. 1038 GLRFGKTRVHYLVLG SGRKSTRVHHWLLVL SGRLSSWVHVGWLVL GLRLKGTRVHYGWVL MRLLKSTLCLVSLCG MRLLKGTLCFVGLCG 6 Phage display (common panning) 3 PMID:9350871 Phosphoenolpyruvate-protein phosphotransferase Phosphocarrier protein HPr 3EZA,3EZB,3EZE,2XDF, Not determined. f3-15mer phage display library (X15) The peptides were synthesized except GLRLKGTRVHYGWVL and MRLLKGTLCFVGLCG. All of them inhibited the phosphotransferase system in vitro. 1039 SDTRGDPVFNLPFQ(7) APVDSVFDRAFSAYL(3) TGLPSSDRAFLVSVS(1) LRDPVFNIAAGVAL(1) LDDPVFGFARRVPV(1) VGSFFALGDRAFLGL(1) MCAVRDPAFSRSRLS(2) LSSARSPRSLDRAFV(2) GGFSSDPGFLLRPRV(1) LMDPVFGFDRRVPF(1) LLDPSFDYFQLYTT(1) DFGPLFSDPGFRSAS(1) GPLDRAFSWRLNPRV(1) RMCSLDLGFCQTILR(1) GVRLLSGVFSDPAFA(1) LMDPVFGFDRRVPV(1) VHLDSAFWFVRTDFD(1) GGFSSDPGFLLRPRG(1) SPDDPAFRFAPYFTP(1) WRTFSRVDRAFFL(1) 20 Phage display (common panning) 3 PMID:9356343 Anti-preS1 monoclonal antibody MA18/7 PreS1 of Large envelope protein Not determined. Not determined. f3-15mer phage display library (X15) All phage mimotopes competed with recombinant HBsAg particles containing the pre-S1 region for binding to MA 18/7. Mouse antisera raised against four mimotopes from the phage display library reacted with HBsAg particles containing pre-S sequences. 1040 HRWWWSPR(2) HRFWWTFR(1) HRWCNFLR(1) KHRWWYQW(1) HKWRWWAF(1) HKKFSSGR(1) HGWLRWFS(1) TNARQTGR(1) AGGGLGRW(1) YHWPAWGQ(1) 10 Phage display (common panning) 3 PMID:9373320 Anti-C5a monoclonal antibody 2925 C5a anaphylatoxin Not determined. Not determined. X8 phage display library MAb 2925 was purified from cell culture supernatant. Biotinylated fabs of mAb 2925 were used in biopanning. The first round of biopanning was performed on 35-mm polystyrene Petri dishes coated with 10 mg streptavidin. In the second and third round the biotinylated Fab fragment was prereacted with phage in solution in a tube. 1041 HRWWWSPR(7) HKWRWWAL(8) HRLYWWRI(3) HRLKWWLG(1) HGRWWLRG(1) YHWAHWVR(1) HSKWWVFR(1) HRWWWSPR(10) HRFWWTFR(1) HKRWYFN(1) HKFRWHR(1) 13 Phage display (common panning) 4 PMID:9373320 Anti-C5a monoclonal antibody 2925 C5a anaphylatoxin Not determined. Not determined. X8 phage display library MAb 2925 was purified from cell culture supernatant. Biotinylated fabs of mAb 2925 were used in biopanning. The first round of biopanning was performed on 35-mm polystyrene Petri dishes coated with 10 mg streptavidin. In the second, third and fourth round the biotinylated Fab fragment was prereacted with phage in solution in a tube. Round four was done using 250 ng/ml Fab 2925 (5 nM). Next day, 400 μl TBS/Tween were added and the whole volume was pipetted onto streptavidin coated plates (15 min at RT). 1042 RAVCSRVIPGCGADSPSPLYLDETRP(3) HHGGGQDYEGGGESLIFHEYVPG(1) NKVRNVHDTVGGGNRLILSEYTI(3) VTPGAMILVEGGVSPKGARCTDL(1) 4 Phage display (common panning) 3 PMID:9373326 Anti-c-myc monoclonal antibody MYC-X-5/1 Myc proto-oncogene protein Not determined. Not determined. TSAR-12 phage display library 1043 PGIIISEEPPWD(1) GHMLPLILDESA(1) GLILSEGEEDVG(1) IVVVPEKLVLSE(1) 4 Phage display (common panning) 3 PMID:9373326 Anti-c-myc monoclonal antibody MYC-X-5/1 Myc proto-oncogene protein Not determined. Not determined. CWl phage display library 1044 RNVPPIFNDVYWIAF(31) RHVAAAVFVGWAFSV(3) TEFLWGFRTVFHG(2) TRWGESDSFRISPPG(1) PGAVRFTFGGSWHY(1) GGWGQFRLFYGAPFD(1) CSSEYGVTYWVLCA(1) IGRIVHHSLYSWPS(1) ECHFLFLLCRVWGR(1) WSVRYDYLVYPSLLP(1) SSGFRDAFRGWDGSA(1) SDVHYIHAHWAVTSH(1) SAVSVLGYHSYFVFP(1) GSFIIFFLVLFMLV(1) PVRYGFSGPRLAILW(1) AARTLSFHPYGYPPY(1) GHGLYYWNFTYSSET(1) TEFLWFRTVLHG(1) LSGGFVWMGFRPSIG(1) RNQGGNWMRFMRCLL(1) VSWSFYRIFGHPGTD(1) LSWSIDYNRNTPSIG(1) 22 Phage display (common panning) 4 PMID:9045678 Caveolin-1 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) The GST-caveolin-1-(61-101) fusion protein was purified and immobilized on glutathione-agarose beads. All phage clones tested only interacted with the peptide that corresponds to the caveolin-scaffolding domain (residues 82-101) and with recombinant full-length caveolin-1 purified from E. coli. 1045 MWHWEKRKWV(13) KWAWGLDRWV(2) HWAWEVRMWR(2) MWRWESCCWE(1) MWVWEHNAWE(1) VWSWALRKWV(1) VWHWAVSRFN(1) MWRWESSRWE(1) RWHWQSHMWL(1) KWLWGSSRWE(1) RDWVGWVCL(1) W-x-W-x(4)-W 11 Phage display (common panning) 4 PMID:9045678 Caveolin-1 Not determined. Not determined. Not determined. [SC]X10[SC] phage display library The GST-caveolin-1-(61-101) fusion protein was purified and immobilized on glutathione-agarose beads. All phage clones tested only interacted with the peptide that corresponds to the caveolin-scaffolding domain (residues 82-101) and with recombinant full-length caveolin-1 purified from E. coli. 1046 GWRPDF(1) MWRPDF(9) WRPDFG(1) WRPFEL(1) 4 Phage display (common panning) 3 PMID:9419019 Anti-FIV monoclonal antibody 3e3 Envelope glycoprotein gp150 Not determined. Not determined. f3-6mer phage display library (X6) Each round was performed using a different solid support. Panning 1: streptavidin magnetic beads (100 μl, Promega) plus 10 μg biotinylated mAb in 100 μl Tris buffered (50 mM) saline (0.15 M) containing 0.1% Tween-20 (TBS-Tw). Panning 2: biotinylated mAb (10 μg) on petri dishes coated with streptavidin (10 μg in 1 ml bicarbonate buffer, 10 M, pH 8.5) and blocked with 1% BSA. Panning 3: 1 ug mAb directly coated on tosyl-activated magnetic beads (Dynal) according to the manufacturer\'s instructions. 1047 YMSVSNSILSWYNSP(1) EDNSPMLMWFNKTWS(1) LKWYNQQF(1) PEVRAQDGLPWYLWK(1) DATDAGWNKLPWYRS(1) SPETPWYQALFYSGR(1) AGSYWCKAWGLDCTS(28) 7 Phage display (common panning) 3 PMID:9430247 Anti-gp120 monoclonal antibody A32 Surface protein gp120, SU Not determined. Not determined. f3-15mer phage display library (X15) 1048 WYFDYHWDRTPISRA RPWNDKSPKNLWYNS GLEGLVPEQMKWYNS RGRKLESSWYNTVWR TESPGWNVPWIAVKW IRHAKVPWYHYPGGR 6 Phage display (competitive panning) 3 PMID:9430247 Anti-gp120 monoclonal antibody 19b Surface protein gp120, SU Not determined. Not determined. f3-15mer phage display library (X15) The HIV-1 gpl20 V3 region peptide CSIHIGPGRAFYTTGC was used in competition for human MAb 19b binding. 1049 SRGFYVQQMHISLES ARSFFIGQANVAMVS FSRNFFFSAGQQVQM MVFMEGNGRSFYYKL 4 Phage display (competitive panning) 3 PMID:9430247 Anti-gp120 monoclonal antibody 19b Surface protein gp120, SU Not determined. Not determined. f3-15mer phage display library (X15) The HIV-1 gpl20 V3 region peptide CYQKRKRIHIGPGRAFYTTC was used in competition for human MAb 19b binding. 1050 WLMRMAAWPWDYTQL NLRSTSFFELWAKWP GEFPDWDNLPLLCEG GRPQWYLQFEDYWRS DSLRVDEHHEVWVPM DWVPFFGFFFSRLQLP NWPRWWEEFVDKHSS FVWEEHVGFLMRN NWPRWEEFVDKHSS CVFYANVEEEVQCWL CLPAYGCKSFREPF GAQVNRKCAWHPRHI VAKKLWVPQVSGSNF HHGVYR 14 Phage display (common panning) 3 PMID:9430247 Anti-gp120 monoclonal antibody b12 Surface protein gp120, SU Not determined. Not determined. f3-15mer phage display library (X15) 1051 NWPRWWEEFVDKHSS(16) 1 Phage display (competitive panning) 3 PMID:9430247 Anti-gp120 monoclonal antibody b12 Surface protein gp120, SU Not determined. Not determined. f3-15mer phage display library (X15) Phages were eluted with recombinant gp120. 1052 HSFGDLSISPNSLTAWPGTLP(3) FCVRLMCSGLIPFFVLCCFFA(2) FSRADFFPLSYSLSSVPSTAL(2) FSFYPPDIVHTTAFSSFVNPVD(1) SSFFYSALTPSFPSPYSQSSR(1) RFFAFPMVCASFFLVAIAFFP(1) PLPPSRFFITVCLTFLFSLSFF(1) VRPCVASLLLFFCLLFLLLPS(1) ICFPFNTRYCIFAMMVSSLVF(1) QQAGSYPGCIDYYYCHASAIG(1) VRFHLAGWLPAVVSFVIFSDH(1) DGLRSDGSWLARFVFNGSGFY(1) FAHEGSASFRLSSKVEDWVSR(1) RHFVPLYLSVSYDGFSRGASI(1) PIIHPHPPRIAMRVSISPFP(1) TPTDSTVRGSSTMDGFLKSVY(1) TASFWRSVSFPWVLSFLAFSH(1) GNFTASTASWGDELLALY(1) 18 Phage display (common panning) 3 PMID:9430247 Anti-gp120 monoclonal antibody b12 Surface protein gp120, SU Not determined. Not determined. X21 phage display library 1053 CSEFHFGPHRGVPRGC CKRIHFGPGRVGGXTC CVRTHFGPGRVMEVVC CLMNHLGPGRSARVDC CRAFHIGPGRGSHRHC CSAHHVGPGRGRVLWC CQGIHYGPGRRSQSC PGMLDGYHYGPGRGS VAMRGVVHHXPGRYV KGEREIVXYGPGRVG CSNFVYGPSRLVQQSC CIGRLYGPGRVTMSGC CFKXFLGPGRVAYVDC CRLVQLGPGRSAAMDC CRAWWIGPGRSGPEAC GTVRPAHVFGPGRGL APVRDRQEFGPGRSR EEHARIRFFGPGRAG WFRRYVLMMGPGRWG CXLIRNGPGRGNTLRC DAVRAVVRWGPGRAG CKILRRGPGXISLEHC AEAPVVVFRGPGRTA CGVVQRGPGRSVMSDC 24 Phage display (common panning) 3 PMID:9430247 Anti-gp120 monoclonal antibody 447-52D Surface protein gp120, SU Not determined. Not determined. CX5GPXRX5C bias phage display library 1054 LLREQRYGPGRHNLHPLL 1 Phage display (common panning) 3 PMID:9430247 Anti-gp120 monoclonal antibody 447-52D Surface protein gp120, SU Not determined. Not determined. LLX5GPXRX5LL bias phage display library 1055 SGSLVEWGCDHKLMC(1) GGTAFPQWGCRELWC(1) EWGCETMRQLCCLPL(2) DRDYFLGWFTTPSIG(4) VGHFGDWFKMPPVGS(2) SWIWGQTFVRAHGRD(1) QDDWVGWWHFMPAK(2) DWSWGLHMTRLG(1) GSGWPWAEA(1) 9 Phage display (common panning) 3 PMID:9430247 Anti-gp41 monoclonal antibody 50-69 Transmembrane protein gp41, TM Not determined. Not determined. f3-15mer phage display library (X15) 1056 CNGRCVSGCAGRC(0.26) CGRECPRLCQSSC(0.15) CGEACGGQCALPC(0.06) NGR 3 Phage display (in vivo) PMID:9430587 Mice carrying human MDA-MB-435 breast carcinoma xenografts Not determined. Not determined. Not determined. CX3CX3CX3C phage display library Endothelial cells in the angiogenic vessels within solid tumors express several proteins that are absent or barely detectable in established blood vessels. CX3CX3CX3C library was screened in mice carrying human MDA-MB-435 breast carcinoma xenografts. Phage displaying CNGRCVSGCAGRC, homed selectively to breast cancer xenografts. This homing can be inhibited by the free CNGRCVSGCAGRC, but not by the CNGRC peptide, even when this peptide was used in amounts 10 times. 1057 CDCRGDCFC(0.80) CTCVSTLSC(0.05) CFRDFLATC(0.05) CSHLTRNRC(0.05) CDAMLSARC(0.05) RGD 5 Phage display (in vivo) PMID:9430587 Mice carrying human MDA-MB-435 breast carcinoma xenografts Not determined. Not determined. Not determined. CX7C phage display library Endothelial cells in the angiogenic vessels within solid tumors express several proteins that are absent or barely detectable in established blood vessels, including alphav integrins and receptors for certain angiogenic growth factors. CX7C library was screened in mice carrying human MDA-MB-435 breast carcinoma xenografts. Phage displaying CDCRGDCFC, homed to several tumor types (including carcinoma, sarcoma, and melanoma) in a highly selective manner, and homing is specifically inhibited by the cognate peptide. The embedded motif RGD has been shown to bind selectively to alphav beta3 and alphav beta5 integrins. 1058 CGSLVRC(0.35) CGLSDSC(0.12) CYTADPC(0.08) CDDSWKC(0.08) CPRGSRC(0.04) GSL 5 Phage display (in vivo) PMID:9430587 Mice carrying human MDA-MB-435 breast carcinoma xenografts Not determined. Not determined. Not determined. CX5C phage libray Endothelial cells in the angiogenic vessels within solid tumors express several proteins that are absent or barely detectable in established blood vessels. CX5C library was screened in mice carrying human MDA-MB-435 breast carcinoma xenografts. The motif GSL and its permutations were frequently recovered from screenings using breast carcinoma, Kaposi's sarcoma, and malignant melanoma, and homing of the phage was inhibited by the cognate peptide. 1059 WHDWAYW(3) LPLWAIV(1) TTWWWQF(1) SAPWWTF(1) LPPWAAF(1) GHTWWTF(1) KPMWWHF(1) SWWSFTP(1) WHSFPPP(1) WHSFPDS(1) WHDFPLV(1) TPTLEAA(1) SPLNTQR(1) SPPSAML(1) QNWWFSF(1) GWYAFTQ(1) SWWDFTQ(1) THLSFLS(1) YHSFNGT(1) WHTFDYS(1) W-x(2)-F 20 Phage display (common panning) 3 PMID:9525900 Protein Vpr Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Purified and denatured His-tag Vpr produced from baculovirus was used by coating plates as the target protein to screen the binding phage for Vpr interactions. Three rounds of panning were carried out at constant stringency. Peptides specific for the His-tag Vpr binding were selected by eluting with 5 mM reduced glutathione in Tris-buffered saline. 1060 TPWWSFM(2) SWWSFYP(2) AWWEFLD(2) QPWWAFF(1) SWWSFSM(1) KWWEFPA(1) TWWGFPA(1) W-x(2)-F 7 Phage display (common panning) 3 PMID:9525900 Protein Vpr Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Native GST-Vpr fusion protein was used as a target protein to screen the binding phage for Vpr interactions through binding to beads. Three rounds of panning were carried out at constant stringency. peptides specific for the GST-Vpr binding were selected by eluting with 5 mM reduced glutathione in Tris-buffered saline. 1061 CLLRMSIC(10) CRLKSRRNC(3) CLPPPNRRC(2) CHTLLKSQC(1) CRRIKPRIC(1) CTMLNRNMC(1) CRLIHLMLC(1) CIRLRXLRC(1) CPISNRLIC(1) CPLMRRILC(1) CRRNRSRKC(1) CRTRPKRRC(1) CKPLMSIRC(1) CRRLRLHRC(1) 14 Phage display (competitive panning) 3 PMID:11532073 Intercellular adhesion molecule 1, ICAM-1 Integrin alpha-L beta-2 1MQ8,3TCX, Not determined. Ph.D.-C7C phage display library (CX7C) Authors used the mAb 84H10 to elute bound phage, because this mAb has previously been shown to bind ICAM-1 and prevent LFA-1 interaction, implying that LFA-1 and 84H10 bind at similar locations on ICAM-1. Fourteen different peptide sequences were identified and analysis of these sequences did not show a consensus sequence. One peptide sequence (*CLLRMRSIC*) was present on 10 of the 26 sequenced phage. 1062 FEGFSFLAFEDFVSSI(2) RDGWYESVSYWGVIDW(2) GAEGIEVKYWVDLGWV(1) FAGAAWHERLGYGHAT(1) PYRIDAWADVDEMVWM(1) IFVGAYVVN(1) FDGFSFLAFEDFVSSI(1) ILGHTWQALGWLVTGR(1) 8 Phage display (common panning) 5 PMID:12963036 Intercellular adhesion molecule 1, ICAM-1 Integrin alpha-L beta-2 1MQ8,3TCX, Not determined. X16 phage display library To assure a native presentation of the extracellular domains of ICAM-1 we have transfected COS-7 cells with plasmid inserted with cDNA encoding the common form of ICAM-1. The obtained ICAM-1 positive cell subline was used for the screening of the phage library. Several clones contained the X1E/DX3X4SX6X7X8X9X10DX12 motif, where the amino acid distribution at X1 and X12 favored aromatic amino acids, while X3, X4, X6 to X10 seem to be random. 1063 FEGFSFLAFEDFVSSI(10) 1 Phage display (subtractive panning) 4 PMID:12963036 Intercellular adhesion molecule 1, ICAM-1 Integrin alpha-L beta-2 1MQ8,3TCX, Not determined. X16 phage display library To assure a native presentation of the extracellular domains of ICAM-1 we have transfected COS-7 cells with plasmid inserted with cDNA encoding the common form of ICAM-1. The obtained ICAM-1 positive cell subline was used for the screening of the phage library. During which the library was first subtracted on the nontransfected COS-7 cells in each of the four rounds of panning prior to exposing the library to ICAM-1-expressing cells. 1064 CSPQYWTGPAC(12)[1.55 ± 0.51, 0.43 ± 0.11] CVQFPTSEKMC(1)[1.50, 0.47] CSDPRKMCIYC(1)[1.86, 0.70] CIWENAGRMVC(1)[1.95, 0.80] CHAGTFLQVAC(1)[1.19, 0.29] CLVAQINLEMC(1)[2.09, 0.88] CERHTKFPSVC(1)[1.92, 0.70] SPQYWTGPA 7 Phage display (common panning) 4 PMID:14728804 CD81 antigen Envelope glycoprotein E2 Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA Phage clones binding to MOLT-4 cells were detected by horseradish peroxydase (HRP)-conjugated anti-M13 phage antibody in the absence or presence of anti-hCD81 MAb by ELISA. The absorbance at 450 nm was measured and average values from two independent experiments were determined. The original phage peptide library PVIII9aaCys without selection was used as a negative control and the corresponding A450 values were 0.2 and 0.17. Data shown were reproduced from the graph. 1065 WTYIPPV[High] WLYVPPV[ND] FTYVPHT[ND] FTYMPPV[High] FTYMPLP[ND] FEWVPMI[ND] FPCFFCY[Very high] PLPISPR[High] ATTLLLA[Low] GSQSHHI[Low] [FYW]-[RNDCEQHKSTY]-[FYW]-[AGILMFPWV]-P 10 Phage display (common panning) 3 PMID:14506253 Chaperone surA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Contents in the square bracket were based on results of ELISA affinity evaluations. ND represented not determined. Mature E. coli SurA protein was expressed as a fusion with a self-cleaving intein. Authors find that both full-length SurA and its core module have similar peptide bind-ing specificities and affinities for peptides with a consensus sequence of the form aromatic-polar-aromatic-nonpolar-proline. 1066 WTYIPPV[High] WEYIPNV[Very high] WSYIPVV[High] FNYVPPT[High] FTYIPNS[High] FTYIPHT[ND] FDFNRRI[High] TAPGVST[Low] [FYW]-[RNDCEQHKSTY]-[FYW]-[AGILMFPWV]-P 8 Phage display (common panning) 3 PMID:14506253 SurA (delta-P2) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Contents in the square bracket were based on results of ELISA affinity evaluations. ND represented not determined. Mature E. coli SurA protein was expressed as a fusion with a self-cleaving intein. Authors find that both full-length SurA and its core module have similar peptide bind-ing specificities and affinities for peptides with a consensus sequence of the form aromatic-polar-aromatic-nonpolar-proline. 1067 FRDIISGTS(7) WKDVVSGRH(2) SIIDGIRAL(1) ERQVITGRV(1) GRDIITGRI(1) ERDIISGIA(1) IGNIITGTK(1) TGNVMTGLK(1) DILDGLK(1) ERDVVTGRA(1) RDIITGLLM(1) DRDIISGRK(1) FKDITSGTS(1) NVITGLRHT(1) TKDIVMGTQ(1) DRNVITGHS(1) ERDVVRGTV(1) NIITGQRQY(1) YRDVLRGVL(1) HRDILRGTA(1) DRNIITGFG(1) YKDPTRGTD(1) 22 Phage display (common panning) 3 PMID:12794137 Anti-b558 monoclonal antibody NL7 Cytochrome b-245 heavy chain Not determined. Not determined. J404 phage display library (X9) Epitope mapping by phage display analysis indicated that NL7 binds the 498 EKDVITGLK 506 region of gp91phox. Each selected peptide shown contains from four to eight residues similar to those of the identified epitope. 1068 FHFNPYTGHPLT(2)[1.09 ± 0.07] KVVPPWHSVLPG(1)[0.05] SPKLLTNWHLLI(1)[0.06] HNWGGSDFWSFG(1)[0.08] FDPNNYWTPMGR(1)[0.08] TDMQFPGFHAAL(1)[0.10] EYPDPPAWHLWF(1)[0.10] WSATWTLSDTWK(1)[0.10] YTTPPFHTGWIW(1)[0.10] YHAGRVNTMSWL(1)[0.10] 10 Phage display (common panning) 4 PMID:12199523 Monoclonal antibody 5E8 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Optical densities at 415 nm were measured with an automated ELISA plate reader. The value of the peptide FHFNPYTGHPLT was expressed as mean ± SD. Data shown were reproduced from the graph. These 11 clones were subjected to analysis by phage ELISA. Only FHFNPYTGHPLT clones were found to be reactive to MoAb 5E8 but not to control anti-FLAG M2 MoAb in the ELISA assay. We compared this peptide sequence with those of dipeptidyl peptidaseIV (DPPIV,E.C.3.4.14.5) and Cell-CAM105, which proteins were located by a database search based on the information of tissue localization and approximate molecular weight of the MoAb 5E8 antigen, and sequence similarity with a region in DPPIV (aminoacids225-233) but not with Cell-CAM105 was found. 1069 MHNHHNHPRPSS[0.56 ± 0.05] MHSDMHAPVSDI[0.91 ± 0.10] HTKPHAWHLMSH[0.25 ± 0.03] HTMYYHHYQHHL[0.99 ± 0.06] LPNPISPRWWVG[0.35 ± 0.02] CDPLLKHHTHPK[0.33 ± 0.11] NHPAVVEPAPSL[0.50 ± 0.03] 7 Phage display (common panning) 4 PMID:12183450 Kinase domain of VEGFR2 Vascular endothelial growth factor A, VEGF-A Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance at 492 nm was measured. Data shown are the mean ± S.D. of three independent experiments and reproduced from the graph. A novel peptide designated K237-(HTMYYHHYQHHL), which was isolated from a phage-displayed peptide library, binded to KDR with high affinity and specificity. By interfering with the VEGF-KDR interaction, the peptide K237 inhibited proliferation of cultured primary human umbilical vein endothelial cells induced by recombinant human VEGF165 in a dose-dependent and cell type-specific manner. 1070 FGGETFTPDWMMEVAIDNE(3) ASSEEETPAWMMEGVFEAW(3) KMGEEFTPDWMMESDFQGW(1) EKQDWYTPEWMMEEGWTVW(1) 4 Phage display (common panning) 6 PMID:12471134 Anti-GXM monoclonal antibody 2H1 Glucuronoxylomannan, GXM Not determined. Not determined. L200 phage display library 1071 KMGGTNHPE(2) KSAVTNHGI(2) HLNHPMSIM(1) KYKFEEVWR(4) KMAFQDVWM(2) KSGFNEVWP(1) K-x(3)-T-N-H-P, K-x(2)-F-x-[ED]-V-W 6 Phage display (in vivo) 3 PMID:11786117 Proximal convoluted tubules, PCT Not determined. Not determined. Not determined. LL9 phage display library (X9) 1072 CKDTSWLGEWLC(4) CEKSSWLAEWLC(2) CLLTSFLGEVYC(1) CLISQRHVASSC(1) C-[TS]-S-W-L-[AG]-E-W-L-C 4 Phage display (in vivo) 3 PMID:11786117 Proximal convoluted tubules, PCT Not determined. Not determined. Not determined. CL10 phage display library (CX10C) 1073 MGSHIEPGG(4) TGSFGVAGG(2) GGMVSQGSK(1) GGMGEHGSS(1) G-[GS]-x(4)-G-[GS] 4 Phage display (in vivo) 4 PMID:11786117 Proximal convoluted tubules, PCT Not determined. Not determined. Not determined. LL9 phage display library (X9) In an attempt to deplete the linear nonapeptide library LL9 of ligands binding unspecifically to different neph-ron segments, we incubated a library aliquot with isolated CCD prior to panning unbound phage on microdissected PCT. 1074 GVKGVQGTL(3) HGVRGNLIS(2) GVRGQLATP(1) GMRDHRMTI(4) ETMQRDVRA(2) RDFRDIWA(1) SLRDRGFT(1) HLNMWRDGG(1) GGAIKDTQN(1) G-V-[KR]-G-x(3)-[TS], R-D-x-R 9 Phage display (in vivo) 4 PMID:11786117 Proximal convoluted tubules, PCT Not determined. Not determined. Not determined. LL9 phage display library (X9) In an attempt to deplete the linear nonapeptide library LL9 of ligands binding unspecifically to different neph-ron segments, we incubated a library aliquot with isolated HEK-293 cells prior to panning unbound phage on microdissected PCT. 1075 CAPGPSKSC(4)[NT] CPHPGTRHC(3)[NT] CSQYPSRSC(2)[NT] CEHFFSRSC(2)[NT] CLNNSQAHC(2)[NT] CNQRHQMSC(1)[0.301, 0.172] CNHRYMQMC(1)[0.351, 0.182] CTPYPSKSC(1)[NT] CLMTPSKRC(1)[NT] CQEPTRLKC(1)[NT] CKEPTRAHC(1)[NT] CTNTGHRHC(1)[NT] CPGKISRSC(1)[NT] CPNNKSASC(1)[NT] CMNQRVQNC(1)[NT] CMNQTPDLC(1)[NT] CTNQFLQQC(1)[NT] CTKMRLEC(1)[NT] 18 Phage display (in vivo) 4 PMID:14578186 Atherosclerotic lesion surfaces Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The synthetic peptides with homology to TIMP-2 bind to endothelial cells in a dose-dependent manner. Binding of peptides (0.5 μmol/L) was inhibited by purified TIMP-2 protein (0.05 μmol/L), indicating that the peptides mimicking TIMP-2 binding are consistent with a novel binding site for TIMP-2 protein on the surface of these cells. The scrambled control peptide did not bind. The absorbance at 405 nm in the absence or presence of TIMP-2 protein was reproduced from the graph and shown. NT represents no tested. Through repeated biopanning, 103 peptidyl sequences were identified, many are homologous to known proteins. 1076 LPALDPTKRWFFETK(21) AHWNSENTVVGLPSK(6) GMQMHQRHVYLSKRP(3) 3 Phage display (common panning) 3 PMID:11836009 Anti-estradiol monoclonal antibody E2-15 Estrogen Not determined. Not determined. X15 phage display library Phage particles was incubated with biotinylated mAb. The preincubated phage-mAb mixture was diluted with PBST and then transferred to the streptavidin-coated plate. More than 70% of the clones displayed the peptide sequence LPALDPTKRWFFETK. There was no consensus among the three peptide sequences. All three phage specifically bound mAb E2-15 in a dose-dependent manner, but failed to bind other tested anti-steroid monoclonal antibodies (anti-progesterone mAb and anti-testosterone mAb). 1077 KGTFDPLQEPRT(1)[1.59 ± 0.09] QDPLFPAMATWA(1)[NT] WDPLLPPFPPPN(1)[NT] SPMDRFDPLHLP(1)[NT] NNPYDALYNFPR(1)[NT] LSKHQFDALLPP(1)[NT] DSMDVFQPPPIW(1)[NT] SLFDVLWSPPPK(1)[NT] DPL, DAL, D-V-[LF] 8 Phage display (common panning) 3 PMID:11425742 Anti-BTx-A monoclonal antibody BT57-1 Botulinum neurotoxin type A, BoNT/A Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance at 490 nm was read with a microplate reader. The synthetic peptide P4M (KGTFDPLQEPRT) corresponded to the phage-displayed peptide selected by BT57-1 and was able to bind the antibodies specifically. Data shown were reproduced from the graph. NT denotes not tested. 1078 MCIDCYLSTLGL(2) YIKPPVSCYDCQ(2) MCWDCTLHARSD(1) CYSMCLDCIMFG(1) C-x-D-C 4 Phage display (common panning) 3 PMID:11425742 Anti-BTx-A monoclonal antibody BT150-3 Botulinum neurotoxin type A, BoNT/A Not determined. Not determined. Ph.D.-12 phage display library (X12) 1079 CKIAK(1)[0.49] CQPMDQACDL(1)[0.46] CQPMEQACDL(1)[0.48] 3 Phage display (common panning) 4 PMID:12191519 Anti-glutamic acid decarboxylase monoclonal antibody GAD-6 Glutamate decarboxylase 2 Not determined. Not determined. C9C T7 phage display library ELISA Absorbance at 405 nm was measured with a microtitre plate reader (Molecular Devices). The mean OD of the antigencoated wells was corrected by subtracting the mean OD of the equivalent blank wells. Data shown were reproduced from the graph. 1080 ARRWDCDGHMCWAQI(3)[1.06 ± 0.04] 1 Phage display (common panning) 5 PMID:12191519 Anti-glutamic acid decarboxylase monoclonal antibody GAD-6 Glutamate decarboxylase 2 Not determined. Not determined. f88-Cys4 phage display library (X4CX4CX5) ELISA Absorbance at 405 nm was measured with a microtitre plate reader (Molecular Devices). The mean OD of the antigencoated wells was corrected by subtracting the mean OD of the equivalent blank wells. Data shown were reproduced from the graph. 1081 LCSTVHCCPGSST(1)[0.56] DPGRSNWSMSFD(1)[1.38] SPSFPLWSFSYL(1)[1.09] 3 Phage display (common panning) 3 PMID:12191519 Anti-glutamic acid decarboxylase monoclonal antibody N-mAb Glutamate decarboxylase 2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance at 405 nm was measured with a microtitre plate reader (Molecular Devices). The mean OD of the antigencoated wells was corrected by subtracting the mean OD of the equivalent blank wells. Data shown were reproduced from the graph. 1082 SFLQTEIDNMGR(1)[1.62] WDLTYELDRLWT(1)[1.06] GFLIWEVDTLSP(1)[1.72] 3 Phage display (common panning) 3 PMID:12191519 Anti-glutamic acid decarboxylase monoclonal antibody N-mAb Glutamate decarboxylase 2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance at 405 nm was measured with a microtitre plate reader (Molecular Devices). The mean OD of the antigencoated wells was corrected by subtracting the mean OD of the equivalent blank wells. Data shown were reproduced from the graph. 1083 AINTICSTPLCWNEA(1)[0.73] ATNRPCSTPMCMGSY(1)[1.24] ASHDSCSTPMCSTPR(1)[1.31] 3 Phage display (common panning) 6 PMID:12191519 Anti-glutamic acid decarboxylase monoclonal antibody C-mAb Glutamate decarboxylase 2 Not determined. Not determined. f88-Cys4 phage display library (X4CX4CX5) ELISA Absorbance at 405 nm was measured with a microtitre plate reader (Molecular Devices). The mean OD of the antigencoated wells was corrected by subtracting the mean OD of the equivalent blank wells. Data shown were reproduced from the graph. 1084 CLASRVSSFLGWGSNLC(5) CSASFFSWLFGWELAC(1) CESFWASRFSSLGGMAC(1) CSDMWLSRLLFTFLGWC(1) CVLVELASLVGLSLWEC(1) CLANGWGCFLGCWFHGC(1) CHVLAMVVGFLGVEWFC(1) FLGW, LASR 7 Phage display (common panning) 5 PMID:12183059 Vascular smooth muscle cells, VSMC Not determined. Not determined. Not determined. CX15C phage display library Phages were added to a monolayer of dedifferentiated, proliferating VSMC. 1085 CNIWGVVLSWIGVFPEC(3) CESLWGGLMWTIGLSDC(3) CSFLTRLGCEIGRFLRC(1) CDCTVRIGRVCMAASC(1) CNGVIFSWGRGWELAC(1) IGR 5 Phage display (in vivo) 6 PMID:12183059 Vascular smooth muscle cells, VSMC Not determined. Not determined. Not determined. CX15C phage display library Before in vivo selections, phages were selected by three rounds of biopanning on cultured VSMC. In this case, non-internalized phages were not removed, to ensure the largest possible variation within this pre-selection. After three rounds of additional in vivo biopanning, 84 clones were picked and grown individually in 96-well plates and tested for binding to VSMC in a phage binding assay. 1086 FEYWTDS(1) SFTYWTN(9) SFTYWSY(5) SFVYWLP(1) SFTYHVP(3) NSFSYWE(2) DSFRYWA(1) FSFSYWT(1) LSFEYYI(1) TSFTYWY(1) ASFSYWS(1) STFHYWG(2) QTFSYWG(1) QTFLYWG(1) TTFHYWS(1) YTFSYWG(4) QTFLYWV(1) MTFEYWP(1) DTFTYWY(1) TTFTYWP(1) TVFTYWT(3) SQSFHYW(1) DSSFMYW(1) YSSFTYW(1) ESTFYYW(1) NETFWYY(1) SDTFWYF(1) DSPFMYW(1) QPAFMYW(1) EAWFSYW(1) [ST]-F-[Tx]-Y-W 30 Phage display (common panning) 3 PMID:12044970 Murine microglial cell line EOC 20 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 1087 QSXAAEGE(3) YPRATTAT(1) DPRSATMT(1) DPRTAAMA(1) DPRIATMS(1) DPRATTST(1) DPRGAAIS(1) DPRSSAMT(1) DPRAVSGT(1) DPRATTTT(1) DPRAATST(1) DPRATPGS(1) DPRINAST(1) DPRSATVS(1) LSXAAEGE(5) VPPSPSNT(1) DPR 16 Phage display (subtractive panning) 3 PMID:11398171 LNCaP prostate carcinoma cells Not determined. Not determined. Not determined. f8-8mer phage display library (X8) The starting library was initially depleted by one round of panning with a primary prostate epithelial cell monolayer followed by three rounds of biopanning with LNCaP cell monolayers. 1088 AGQGSQ(10)[0.49] LLVPLR(6)[0.53] VYMSPF(2)[0.54] FAYLLG(1)[0.66] VTEMRH(1)[0.30] DHKMRH(1)[0.07] SLHHAS(1)[0.31] VKCSPF(1)[0.42] 8 Phage display (common panning) 4 PMID:12440521 Fibroblast growth factor receptor 1, FGFR-1 Not determined. Not determined. Not determined. f3-6mer phage display library (X6) ELISA Absorbance at 450 nm was measured. Data shown were reproduced from the graph. To retain the natural conformation of FGFR1 during screening, we expressed recombinant FGFR1 on the surface of Sf9 insect cells. 1089 CPVDFDFLC CPRDFEFLC CPADFEFLC 3 Phage display (common panning) 2 PMID:12446478 Anti-PSA monoclonal antibody 5E4 PSA-α2M complexes Not determined. Not determined. CX7C+CX8C+CX10C+CX3CX3CX3C+CX3CX4CX2C phage display library pool 1090 CPSVDGGWTC CKSMDGGWTC CHSACSKHCFVHC 3 Phage display (common panning) 3 PMID:12446478 Anti-PSA monoclonal antibody 5E4 and H117 PSA-α2M complexes Not determined. Not determined. CX7C+CX8C+CX10C+CX3CX3CX3C+CX3CX4CX2C phage display library pool The primary library was panned for two rounds using MAb 5E4, after which the eluate was amplified and panned for one round with MAb H117. 1091 CTWHWSPEEC 1 Phage display (common panning) 2 PMID:12446478 Anti-PSA monoclonal antibody H117 PSA-α2M complexes Not determined. Not determined. CX7C+CX8C+CX10C+CX3CX3CX3C+CX3CX4CX2C phage display library pool 1092 CPSVDGGWTC CHSACSKHCFVYC CHSACSKHCFVHC 3 Phage display (common panning) 3 PMID:12446478 Anti-PSA monoclonal antibody 5E4 and H117 PSA-α2M complexes Not determined. Not determined. CX7C+CX8C+CX10C+CX3CX3CX3C+CX3CX4CX2C phage display library pool The primary library was panned for two rounds using MAb H117, after which the eluate was amplified and panned for one round with MAb 5E4. 1093 CRPWGSSRC CKSWGSSRC CNLYKVGC CHPYKVGC 4 Phage display (common panning) 3 PMID:12446478 Anti-PSA monoclonal antibody 5A10 and 4G10 PSA-α2M complexes Not determined. Not determined. CX7C+CX8C+CX10C+CX3CX3CX3C+CX3CX4CX2C phage display library pool The primary library was panned for two rounds using MAb 5A10, after which the eluate was amplified and panned for one round with MAb 4G10. 1094 CSVYPFWHC 1 Phage display (common panning) 2 PMID:12446478 Anti-PSA monoclonal antibody 4G10 PSA-α2M complexes Not determined. Not determined. CX7C+CX8C+CX10C+CX3CX3CX3C+CX3CX4CX2C phage display library pool 1095 CRSYPFWMC CVSYPFWKC CAVFPFWRC CVWWWGC CAPWWGC 5 Phage display (common panning) 3 PMID:12446478 Anti-PSA monoclonal antibody 5A10 and 4G10 PSA-α2M complexes Not determined. Not determined. CX7C+CX8C+CX10C+CX3CX3CX3C+CX3CX4CX2C phage display library pool The primary library was panned for two rounds using MAb 4G10, after which the eluate was amplified and panned for one round with MAb 5A10. 1096 CSGIAPWLC CGPGIDSWVC CGPGIRSWVC CDYMPLVDNC 4 Phage display (common panning) 2 PMID:12446478 Anti-PSA monoclonal antibody H50 PSA-α2M complexes Not determined. Not determined. CX7C+CX8C+CX10C+CX3CX3CX3C+CX3CX4CX2C phage display library pool 1097 CEGIAVWLC CTRYSGYWVC CTQMNGYWIC CLYDHLC CILLYNDCC 5 Phage display (common panning) 3 PMID:12446478 Anti-PSA monoclonal antibody H50 and 9C5 PSA-α2M complexes Not determined. Not determined. CX7C+CX8C+CX10C+CX3CX3CX3C+CX3CX4CX2C phage display library pool The primary library was panned for two rounds using MAb H50, after which the eluate was amplified and panned for one round with MAb 9C5. 1098 CFFGNWNFC 1 Phage display (common panning) 2 PMID:12446478 Anti-PSA monoclonal antibody 9C5 PSA-α2M complexes Not determined. Not determined. CX7C+CX8C+CX10C+CX3CX3CX3C+CX3CX4CX2C phage display library pool 1099 CWNWNLHISC CYFKNWNFC CVRSCISDCELRC 3 Phage display (common panning) 3 PMID:12446478 Anti-PSA monoclonal antibody H50 and 9C5 PSA-α2M complexes Not determined. Not determined. CX7C+CX8C+CX10C+CX3CX3CX3C+CX3CX4CX2C phage display library pool The primary library was panned for two rounds using MAb 9C5, after which the eluate was amplified and panned for one round with MAb H50. 1100 CWSFFSNIC(26) CWPFWGPWC(4) CRPFWGPWC(4) CQPWHWPPC(2) CWSFFKLYC(2) CNELRSGLC(1) CNSVRVYSC(1) CDILHYSKC(1) 8 Phage display (common panning) 3 PMID:12436474 Capsid protein Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 65% phage clones carried the fusion peptide C-WSFFSNI-C. The N- and C-terminal residues represent constrained cysteine residues. 20% phage clones exhibited the sequence motif C-W/RPFWGPW-C. 1101 QLTMRLQ YGRETST SAESRHY M-x(2)-P 3 Phage display (competitive panning) 1 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1102 RPLPYPD LASPNGH GAASTRY M-x(2)-P 3 Phage display (competitive panning) 2 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1103 MANPGRA MQPSLVH MTTYPLS MTTTMQL MAVDPLR M-x(2)-P 5 Phage display (competitive panning) 3 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1104 MQSPPFH MTPPASI MTYPREF SARPVPL SPRPYST M-x(2)-P 5 Phage display (competitive panning) 4 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1105 MQLPPDT MQTPLAP MSLPLLP MTAPPAR MGNPINH MQISGHP LTRPVSP LAAPPKA DQSRMPT LAKVPPQ M-x(2)-P 10 Phage display (competitive panning) 5 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1106 QNLRGDS EAPAQLP AGPTWAH M-x(2)-P 3 Phage display (competitive panning) 1 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1107 MATPRNS MAARAPM AAAHALF M-x(2)-P 3 Phage display (competitive panning) 2 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1108 MQLPKSL MGKPLHS MSTPSKV LAQFPSP APFTHLR M-x(2)-P 5 Phage display (competitive panning) 3 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1109 MTLPMLQ MSQPPLA MWPTITP TALSRPP SATNQAP M-x(2)-P 5 Phage display (competitive panning) 4 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1110 MQLPVYP MQMPAVN MQPPGSI MATPGLG MAVPQPI MSARFPP LTRPPWG FTKTTSP GPTRILK VRWEMNL M-x(2)-P 10 Phage display (competitive panning) 5 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1111 TRLPHNP RYLVESP WPIRPLS M-x(2)-P 3 Phage display (competitive panning) 1 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1112 MMLHPYA LPSKLLD LVTAPRL M-x(2)-P 3 Phage display (competitive panning) 2 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1113 MAAAPFS MATASQL AAMPIIP LTTVPPY FAKDYSR M-x(2)-P 5 Phage display (competitive panning) 3 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1114 DNPRGFS FHPLNLS NNMAPHL VNRLSEN M-x(2)-P 4 Phage display (competitive panning) 4 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1115 MQLPLAT MGLPLGK MGLPPSP MTLPAAS MTSPPTP MSLSPPQ MGTLPTT LTLPPSP LSPPPTH STHPLQT M-x(2)-P 10 Phage display (competitive panning) 5 PMID:12032665 Fibroblast growth factor receptor 2, FGFR-2 Heparin-binding growth factor 2 (HBGF-2) 1EV2,1II4,1IIL, Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with 911 human embryonic retinoblast cells for 1 hour at 4℃. Then medium containing unbound phage was removed and discarded. Finally, phage bound to cells were specifically eluted by the addition of FGF2 (in PBS) to each well with gentle agitation. 1116 CCAVLNGAY(1) EGFGSGGAI(2) SQNSAEGAW(1) LTHCGSGAS(1) YCHTFHGAG(1) YHANSQGAV(1) EMLQGAGAM(1) PSMSLFGAF(1) EVLSHAGAH(1) GLRPGEGAY(1) IISCDEGVG(1) GLWWAEGVV(1) IVYVGDGVC(1) NLSVEGGVW(1) ARSLFGVD(1) GGFRIFGIV(1) GAGIGWGID(1) CVVGCGGGV(1) G-A-x 18 Phage display (common panning) 3 PMID:11983234 Alzheimer's disease specific monoclonal antibody MN423 Not determined. Not determined. Not determined. X9 λ phage display library 1117 NMMRFTSQPPNN(9)[0.40] NMMKYISPPIFL(9)[1.11] NMMNYIMDPRTH(3)[2.52] NMMRFTELSTPS(1)[0.99] 5 Phage display (common panning) 3 PMID:12682274 Anti-LOS polyclonal antibody Lipooligosaccharide, LOS Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding reactivity was determined by an ELISA, in which each peptide was covalently coupled to NucleoLink strip as a coating Ag, followed by incubation with the target rabbit anti-LOS Ab. The binding reactivity of the synthetic peptides to the target Ab was evaluated by average OD405 values of each diplicate sample after incubation with a goat anti-rabbit IgG alkaline-phosphatase conjugate, and then addition of a substrate. 22 clones were subjected to DNA sequencing, and four consensus sequences were identified as NMMRFTSQPPNN, NMMNYIMDPRTH, NMMKYISPPIFL, and NMMRFTELSTPS. Three of the four synthetic peptides showed strong binding reactivity to the rabbit anti-LOS Ab and also a mouse bactericidal monoclonal anti-LOS Ab in vitro, and elicited specific serum anti-LOS Abs in rabbits (27- to 81-fold) after conjugation with keyhole limpet hemocyanin. 1118 SWTPDKWLRPSG(2)[3.14/3.03] NIAHPDRWDRLR(1)[3.14] ADNPIKWDRART(1)[3.26] LSNTANPWSWDR(1)[3.02] ASNWTDFRVTDR(1)[2.97] GMLPPQLFDRHH(1)[2.91] AEPFYSWDRASR(1)[3.07] P-x(2)-W-D-R 7 Phage display (subtractive panning) 4 PMID:12902494 Monoclonal antibody HC-10 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Reactivity of phage particles with mAb was measured by ELISA. Absorbance was read at 492 nm with a Multiscan microplate reader (Benchmark; Bio-Rad). At each round, phage particles binding to isotypic and allotypic determinants of mAb HC-10 were removed by a preadsorption step on mAb HC-10 isotype-matched mAb W6/32. 1119 HYQHNTHHPSRW(2)[3.9 ± 0.7] HFQAQMRHGHGH(1)[3.6 ± 0.5] HQSHHYGPRDHT(1)[3.4 ± 0.4] HIKHHPSSVPHA(1)[3.2 ± 0.3] GSPHHNHFKESH(1)[3.0 ± 0.1] HSPHIWSPHHGP(1)[3.0 ± 0.3] HLQIPKPHVHHT(1)[2.9 ± 0.5] HSVHHLPSPLSH(1)[2.8 ± 0.2] HPHQAHPSPKAH(1)[2.5 ± 0.3] HSPLGSHHHPKH(1)[2.5 ± 0.2] HNHAHLPLHPAP(1)[2.5 ± 0.6] HNHHPHSTRQAS(1)[2.4 ± 0.3] GTHVHHPHSTST(1)[2.4 ± 0.7] HMTELHHSGVHQ(1)[1.8 ± 0.4] 15 Phage display (common panning) 4 PMID:14644435 Zinc ion, Zn(2+) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Relative binding affinity of phage at 1 nM for metal ions was determined by ELISA. Absorbance was measured at 492 nm. The absorbance ratio of each clone to the primary phage library was estimated. All 14 peptide sequences contained four or five histidines, however, there was no obvious consensus in motif among the selected sequences. 1120 CWKLLGSEEEC(15) CPCFLLGCC(15) CWHKDLLGC(4) CWSMELLGC(1) CPPDLFWYC(4) CPEDLYFFC(3) CPEDFIFFC(1) L(2)-G 7 Phage display (common panning) 5 PMID:11381078 Integrin alpha-M-beta-2, integrin αMβ2 Not determined. Not determined. Not determined. CX7C and CX9C phage display library pool LLG sequences exposed on ICAM-1 and on von Willebrand factor at sites of vascular injury play a role in the binding of leukocytes, and CPCFLLGCC and peptidomimetics derived from it could provide a therapeutic approach to inflammatory reactions. 1121 ELRGDMAAL(3) SLRGDRAGW(2) ELRGDRAHW(1) YRDFRDIWA(1) E-L-R-G-D-[RM]-A-x-[WL] 4 Phage display (common panning) 3 PMID:11158220 Cortical collecting duct, CCD Not determined. Not determined. Not determined. LL9 phage display library (X9) 1122 ELRGDMAAL(5) ELRGDMAHW(2) LRGDAFSLA(1) E-L-R-G-D-[RM]-A-x-[WL] 3 Phage display (common panning) 4 PMID:11158220 Cortical collecting duct, CCD Not determined. Not determined. Not determined. LL9 phage display library (X9) 1123 ELRGDRAHW(6) EIRGDRAHW(1) EMRGDLAGF(1) E-L-R-G-D-[RM]-A-x-[WL] 3 Phage display (common panning) 3 PMID:11158220 Cortical collecting duct, CCD Not determined. Not determined. Not determined. LL9 phage display library (X9) 1124 KMGGTNHPE(4) MKTASVRHP(1) FTNHVSTRR(1) VLNHEMSIM(1) KMAGTNHPS(1) K-x(3)-T-N-H-P 5 Phage display (common panning) PMID:11158220 Proximal convoluted tubules, PCT Not determined. Not determined. Not determined. LL9 phage display library (X9) 1125 RRLGRQTYDNES HDEGRQIIQFEE LRNCEQDFFTLN AFAQAPTHQLSL ESNPVDGAHLSL PNEPDDLALMRIIRI 6 Phage display (common panning) 4 PMID:11874892 Anti-C.pneumoniae monoclonal antibody 8A6 Chlamydia pneumoniae Not determined. Not determined. f3-15mer phage display library (X15) The selected peptide mimotope sequences tended to be composed of charged residues surrounding a core of hydrophobic residues. 1126 CDSAFVTVDWGRSMSLC(11) CVVLHVPHGAEAGLPLY(4) 2 Phage display (common panning) 4 PMID:15203927 Human submucosal-gland carcinoma, Calu-3 cells Not determined. Not determined. Not determined. CX15C phage display library Biopan the phage library on differentiated Calu-3 cell monolayers, the membrane associated phage were degraded by proteinase K digestion and the internalising phage were protected from intracellular degradation by the endosomolytic compound chloroquine, amplified from cell lysates and used in the next round. 1127 FEKYYYW(25)[3.762 ± 0.144] TDRPALS(8)[0.752 ± 0.054] VMQATHD(5)[0.612 ± 0.078] WHTNYEP(3)[0.603 ± 0.065] KVFNWPW(3)[1.891 ± 0.047] YNPLPGT(3)[0.716 ± 0.064] APLESNW(2)[0.672 ± 0.044] LSLPKLP(2)[2.003 ± 0.047] KLWVIPQ(2)[2.081 ± 0.027] LQPMWFA(1)[3.491 ± 0.135] LTYRISP(1)[1.828 ± 0.066] FQNNLQL(1)[1.335 ± 0.039] GSFYVFP(1)[1.185 ± 0.046] LVYPPMV(1)[1.423 ± 0.025] ITPYTVL(1)[1.301 ± 0.049] LPPLNYY(1)[0.801 ± 0.073] HYTSATL(1)[0.552 ± 0.022] WQPNTRP(1)[0.728 ± 0.031] IPRTYPL(1)[0.625 ± 0.010] SAPGLLH(1)[0.615 ± 0.038] DYHNHLT(1)[0.586 ± 0.051] SPTQSTL(1)[0.580 ± 0.041] AQAIMQY(1)[0.517 ± 0.051] HHVKFQN(1)[0.623 ± 0.064] SLCSVLC(1)[0.580 ± 0.012] HTTYFPM(1)[0.552 ± 0.024] HVLPPLH(1)[0.810 ± 0.056] LHAWQDL(1)[0.773 ± 0.045] TGNPPPN(1)[0.745 ± 0.070] LSNYTRP(1)[0.712 ± 0.038] HTPLATA(1)[0.718 ± 0.037] HMPNASF(1)[0.697 ± 0.043] YNPYTPL(1)[0.670 ± 0.046] STLLPES(1)[0.690 ± 0.061] NVNLLLP(1)[0.635 ± 0.022] QANPLMI(1)[0.636 ± 0.019] NTTTYPT(1)[0.525 ± 0.038] 37 Phage display (common panning) 4 PMID:12486713 Cell division protein ftsA Cell division protein ftsA Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Absorbance at 450 nm was measured. Data shown were average ELISA values (OD450nm) ± standard deviation (n = 4). The isolated peptides defined a degenerate consensus sequence, which in turn displayed a striking similarity with residues 126-133 of FtsA itself. This result suggested that residues 126-133 were involved in homodimerization of FtsA. 1128 VHALES DRVTLG LGGLSA CGLSLK GLSARH SKHALE VFALEG EVISGL LISGVL ELLSGL GRLSGSL AMSAGV G(2)-L-S-G-L, HALE, ISGL, MSAG, R-V-T-x-G 12 Phage display (subtractive panning) 3 PMID:12629410 Human urothelial cells Not determined. Not determined. Not determined. X6 phage display library Each library was incubated with MOLT-4 leukemia cells for 30 minutes on ice to deplete background and common cell surface binding phage (pre-clearing). 1129 SAHGTSTGVRGP(4) SVHMINTSQVHL(1) SDTTMGQVHRHP(1) KQASNLIVMHYP(1) YSLPQRTHLHLP(1) HLPTSSLFDTTH(1) HQNLNLGSSWST(1) 7 Phage display (common panning) 4 PMID:11478946 Allergen-specific IgE-mimicking monoclonal antibody 2A1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1130 VDLPEHGK(9) VGLPEHTQ(5) VGLPEHSA(4) VDLPTHSS(7) VDLPEHRQ(1) VDLPTHQS(1) VDLPTHNQ(1) VDLPQHGQ(1) DTTKNGSG(1) V-[DG]-L-P-[ET]-H 9 Phage display (subtractive panning) 4 PMID:14617786 RG2 rat glioma cells Not determined. Not determined. Not determined. f8-8mer phage display library (X8) RG2 cells were grown to a sub-confluent monolayer. First, an aliquot of the primary library in washing/blocking buffer was added to an empty flask (depletion flask) to remove plastic-binding phage clones. Buffer containing phage that did not bind to plastic was transferred to the flask with RG2 glioma cells for incubation. After that, phage not associated with tumor cells were washed away. Cell-surface bound phage were recovered with low pH elution buffer. 1131 ELRGDSLP(10) EVRGDSLP(2) VDLPSHPE(1) VNLPEHPE(2) VDLPRSDT(1) HTTKEQMA(1) V-[DG]-L-P-[ET]-H 6 Phage display (subtractive panning) 4 PMID:14617786 RG2 rat glioma cells Not determined. Not determined. Not determined. f8-8mer phage display library (X8) RG2 cells were grown to a sub-confluent monolayer. First, an aliquot of the primary library in washing/blocking buffer was added to an empty flask (depletion flask) to remove plastic-binding phage clones. Buffer containing phage that did not bind to plastic was transferred to the flask with RG2 glioma cells for incubation. After that, phage not associated with tumor cells were washed away. Cell-surface bound phage were recovered with lysis buffer. 1132 VDLPQHGG(4) VDLPTHTS(1) VNLPEHAQ(2) VGLPEHQP(1) DTTKTSAG(1) DSTKIGTS(1) DSTKASDA(1) DTTQSMHT(1) DSTKSTNS(1) DSTKAVAL(1) DSTKSGNM(1) DTTKGPGT(1) DGTKMAGG(1) V-[DG]-L-P-[ET]-H 13 Phage display (subtractive panning) 4 PMID:14617786 RG2 rat glioma cells Not determined. Not determined. Not determined. f8-8mer phage display library (X8) RG2 cells were grown to a sub-confluent monolayer. First, an aliquot of the primary library in washing/blocking buffer was added to an empty flask (depletion flask) to remove plastic-binding phage clones. Before incubation with RG2 cells, preselection steps with fibroblasts, myoblasts, and hepatocytes were included in the protocol to remove phage binding to these normal cells. Phage associated with RG2 cells were recovered from a single flask with elution buffer. 1133 DTTKGGNP(1) DDTKHSLP(1) DTTRTHMP(1) DSTRGSPA(1) DSTRTTSA(1) DTTRLSDQ(1) DNTRVAAP(1) DDTRYSSA(1) DETLYGIS(1) DYDMTKNT(1) D-[TS]-T 10 Phage display (subtractive panning) 4 PMID:14617786 RG2 rat glioma cells Not determined. Not determined. Not determined. f8-8mer phage display library (X8) RG2 cells were grown to a sub-confluent monolayer. First, an aliquot of the primary library in washing/blocking buffer was added to an empty flask (depletion flask) to remove plastic-binding phage clones. Before incubation with RG2 cells, preselection steps with fibroblasts, myoblasts, and hepatocytes were included in the protocol to remove phage binding to these normal cells. Phage associated with RG2 cells were recovered from a single flask in two sequential steps, first with elution buffer and then with lysis buffer. 1134 DLTKSTAP(9) DTTKSTTT(1) EPVQPHST(1) D-[TL]-T-K 3 Phage display (subtractive panning) 6 PMID:14617786 RG2 rat glioma cells Not determined. Not determined. Not determined. f8-8mer phage display library (X8) RG2 cells were grown to a sub-confluent monolayer. First, an aliquot of the primary library in washing/blocking buffer was added to an empty flask (depletion flask) to remove plastic-binding phage clones. Before incubation with RG2 cells, preselection steps with fibroblasts, myoblasts, and hepatocytes were included in the protocol to remove phage binding to these normal cells. Phage associated with RG2 cells were recovered from a single flask with elution buffer. 1135 ESRGDSYA(2) DLTKSSAP(1) DTTKLTND(1) DNAIYTYQ(2) D-[TL]-T-K 4 Phage display (subtractive panning) 6 PMID:14617786 RG2 rat glioma cells Not determined. Not determined. Not determined. f8-8mer phage display library (X8) RG2 cells were grown to a sub-confluent monolayer. First, an aliquot of the primary library in washing/blocking buffer was added to an empty flask (depletion flask) to remove plastic-binding phage clones. Before incubation with RG2 cells, preselection steps with fibroblasts, myoblasts, and hepatocytes were included in the protocol to remove phage binding to these normal cells. Phage associated with RG2 cells were recovered from a single flask in two sequential steps, first with elution buffer and then with lysis buffer. 1136 QQGRGDIDSR(3) AQCRSDSPSC(3) RQGRGDQRST(3) TQRGDTIDSK(1) VQRGDAPNSS(1) AQRGDSYTSE(1) VQRGDEVLSK(1) AQRGDQLDSY(1) RQRGDGPYSQ(1) RGD 9 Phage display (common panning) 3 PMID:12646694 Integrin alpha-V-beta-3, integrin αvβ3 Not determined. Not determined. Not determined. Tendamistat loop I phage display library (xQxxxxxxSx) 1137 RARGDNPSNL(15) 1 Phage display (common panning) 3 PMID:12646694 Integrin alpha-IIb-beta-3, integrin αIIbβ3 Not determined. Not determined. Not determined. RGDX phage display library (xxRGDxxxxx) 1138 GARGDGNPWE(1) ASRGDRPQDM(1) TSRGDHPRTQ(1) GSRGDSLIMH(1) ASRGDNPLLW(1) TSRGDNPQTM(1) VGRGDSPRNA(1) VTRGDNYYPM(1) ASRGDTPRSY(1) ASRGDTYTTW(1) ATRGDDPRPR(1) VSRGDMPPMW(1) ATRGDEYQPR(1) VSRGDSYIGE(1) VTRGDTYWRK(1) 15 Phage display (common panning) 3 PMID:12646694 Integrin alpha-V-beta-3, integrin αvβ3 Not determined. Not determined. Not determined. RGDX phage display library (xxRGDxxxxx) 1139 ARRGDTFSLL(1) VSRGDTYVES(1) VTRGDTFTQS(1) VARGDTYVAS(1) ATRGDYYVAS(1) RSRGDTPARF(1) VRRGDVPFSS(1) ATRGDTILYL(1) VTRGDTFTIS(1) ATRGDTIEYE(1) AQRGDTVEYF(1) FARGDSLPEY(1) VTRGDSISYL(1) GSRGDTFRLV(1) VSRGDTFSLI(1) 15 Phage display (common panning) 3 PMID:12646694 Integrin alpha-V-beta-5, integrin αvβ5 Not determined. Not determined. Not determined. RGDX phage display library (xxRGDxxxxx) 1140 RKSRGDMGIY(2) HRTRGDMGSY(2) RYTRGDLGLT(1) RHSRGDMGSS(1) HRSRGDMGYW(1) RKSRGDAGLM(1) RTGRGDMGAN(1) RRTRGDLGLV(1) RSSRGDLGLM(1) RMSRGDLGLY(1) RKSRGDYGFV(1) RRSRGDTGIV(1) RTSRGDLGTF(1) 13 Phage display (common panning) 3 PMID:12646694 Integrin alpha-IIb-beta-3, integrin αIIbβ3 Not determined. Not determined. Not determined. XRGD phage display library (xxxRGDxxxx) 1141 HHARGDKDHM(1) ALGRGDSDLI(1) GSGRGDADLH(1) HPARGDSDFE(1) RAARGDSDHR(1) RHSRGDSDHL(1) RPSRGDTDHY(1) RHARGDFDHA(1) RVPRGDSDLT(1) RPGRGDSDQY(1) RQGRGDSRTS(1) RHARGDNDTR(1) RPSRGDMDHM(1) QPARGDMDLR(1) RSARGDSDHR(1) 15 Phage display (common panning) 3 PMID:12646694 Integrin alpha-V-beta-3, integrin αvβ3 Not determined. Not determined. Not determined. XRGD phage display library (xxxRGDxxxx) 1142 HLARGDDLTY(15) 1 Phage display (common panning) 3 PMID:12646694 Integrin alpha-V-beta-5, integrin αvβ5 Not determined. Not determined. Not determined. XRGD phage display library (xxxRGDxxxx) 1143 STHEETP(6) STHEETS(5) AYHTFTP(2) STHHSTP(2) SPHEETP(1) ATHEETP(1) SPHESYP(1) SPHESYL(1) SGRMMRL(1) WEFTFPP(1) STHEEHP(5) STPHSTP(3) STAEEHP(2) SPPEETP(3) ATAHSHP(1) STHHSHP(1) TQPHHTP(1) SMYQPPM(1) TPPHSTP(1) SPPYLTN(1) S-T-x(4)-P 21 Phage display (competitive panning) 3 PMID:12388644 Peripheral-type benzodiazepine receptor Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In fact, isolated mitochondria from the MA-10 mouse Leydig cell tumor line, which express high amounts of PBR, was the target. In biopanning experiments, PBR-interacting phage peptides were eluted using either PK 11195 or Ro5-4864. 1144 SLWPPVS(44) MSPILSP(26) ITQPSLR(8) FKSLPMP(8) GRPLPPS(5) QSTQSPL(5) AHNSAAD(5) AEGPPNE(4) FGSHASE(3) SAALPAR(3) NWLPTPP(2) LVSTPLP(2) KVPWISW(2) SARLPAR(2) VAKIWVD(2) TSTTHTP(2) GLSPATS(2) MSKPTPL(1) IQPFTLQ(1) SAGLPAR(1) IVYRTSS(1) TYQRALY(1) NRTMAPW(1) RSMQMPP(1) TMSLPHH(1) 25 Phage display (common panning) 3 PMID:11994302 Chaperone protein hscA Iron-sulfur cluster assembly enzyme ISCU, mitochondrial Not determined. Not determined. Ph.D.-7 phage display library (X7) Screening of a heptameric peptide phage display library revealed that Hsc66 prefers peptides with a centrally located Pro-Pro motif. 1145 GNNSVSKEKPPSLNWP(1) TELKLAPPVLNAPPL(2) KAHPPLLLNSPRDVPL(1) YPKESPPRLNAPWYQ(1) ESKLTPPPLNPIRVV(2) KPRDTLPPPLNRPPP(3) VPVDAPHAGTKPHSA(2) K-x(2)-P(2)-x-L-N-x-P 7 Phage display (common panning) 3 PMID:11139210 Serum antibodies with Lyme disease Not determined. Not determined. Not determined. f88-15mer phage display library (X15) First-round affinity selections were carried out in four polystyrene 24-well dishes. Wells were coated with streptavidin. The second round was carried out in four 96-well ELISA dishes. A key difference was that neutravidin was substituted for streptavidin to capture Bio-IgG molecules. The third round of affinity selection was like the second round, but streptavidin was substituted for neutravidin. 1146 PKSSCTQNPILCAILS(1) 1 Phage display (common panning) 3 PMID:11139210 Serum antibodies with Lyme disease Not determined. Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) 1147 SCPEGSKLCI(1) TCPEGAKLCD(1) SCAEGAKYCL(1) ACVAGDATCK(1) 4 Phage display (common panning) 3 PMID:11139210 Serum antibodies with Lyme disease Not determined. Not determined. Not determined. f88-LX6 phage display library (XCX6CX) 1148 EKRFACKPLCNTPA(2) 1 Phage display (common panning) 3 PMID:11139210 Serum antibodies with Lyme disease Not determined. Not determined. Not determined. f88-Cys3 phage display library (X5CX3CX4) 1149 QKDFACKHCKLPSP(2) RPDRLCPCVDPRE(1) K-x-F-A-C-K 2 Phage display (common panning) 3 PMID:11139210 Serum antibodies with Lyme disease Not determined. Not determined. Not determined. f88-Cys2 phage display library (X5CX2CX5) 1150 QRSECSTSKCFVRK(2) YREACTNGKCFVLK(2) R-x(2)-C-x(3)-K-C-F-V-x-K 2 Phage display (common panning) 3 PMID:11139210 Serum antibodies with Lyme disease Not determined. Not determined. Not determined. f88-Cys4 phage display library (X4CX4CX5) 1151 CSDTRRGDC(23)[0.80] CKLGPIRGC(5)[0.60] CLTSHNMMC(3)[0.25] CPWTKAYHC(3)[NT] CNSYSLSRC(2)[NT] CRSHTPRSC(1)[NT] 6 Phage display (common panning) 3 PMID:14631075 Beta-lactamase TEM Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance at 405 nm was measured. Data shown were reproduced from the graph. NT denotes not tested. 1152 PLFYSV KIPRTLT PLRLSW PRAVST KGPRQIT PRPLSG WPLGLA FRPRSIT RLPVGLT RSPKSLT PVWLAA IHPSSLTA PASFTS GQPHYLT MKPASWT TPAYMLT PLYLT PGLIGT PRSISN RLPASYT NPPRYLT PKTQIS SVPRHFT LLPAWLT THPYTMT LRPAKST SGPSTST GSGLKA AMGLKS KVGLRT GRRLIHH HPRRSIT GRRLLSR ALRRLET FYKRVLT FRRICV VFFRRQTA GLARNITA FGSRYLTA QDRYLNT 40 Phage display (common panning) 3 PMID:11279151 Matrix metalloproteinase-9, MMP-9 Not determined. Not determined. Not determined. X6 phage display library 1153 QHFNNSVNLGFT(11) SPERHLHDLRPY(1) NKLYPDAYFNPG(1) GGNWTAATASWA(1) RICMMLVIRSNA(1) 5 Phage display (subtractive panning) 2 PMID:11121403 BAG family molecular chaperone regulator 1 Heat shock cognate 71 kDa protein 1HX1, Not determined. Ph.D.-12 phage display library (X12) The recombinant Hap46 was immobilized on Petri dishes, blocked, and incubated with phage library following the manufacturer's protocol. To eliminate enrichment of phage clones interacting with the plastic surface or the blocking agent, the following two strategies were employed: (i) eluted and amplified material from the first panning was either subjected to a preadsorption step on bovine serum albumin coated Petri dishes prior to subsequent pannings, or (ii) bovine serum albumin as blocking agent was alternated with casein in the form of dried milk powder. 1154 QHFNNSVNLGFT(20) SPERHLHDLRPY(3) AILIISLMLGFT(1) APTVKNCPSPCP(1) YTTTWRPPYSHN(1) YWWIIIRQYTTP(1) TLATVPSSSLFV(1) LVDHHQTYTTPP(1) 8 Phage display (subtractive panning) 3 PMID:11121403 BAG family molecular chaperone regulator 1 Heat shock cognate 71 kDa protein 1HX1, Not determined. Ph.D.-12 phage display library (X12) The recombinant Hap46 was immobilized on Petri dishes, blocked, and incubated with phage library following the manufacturer’s protocol. To eliminate enrichment of phage clones interacting with the plastic surface or the blocking agent, the following two strategies were employed: (i) eluted and amplified material from the first panning was either subjected to a preadsorption step on bovine serum albumin coated Petri dishes prior to subsequent pannings, or (ii) bovine serum albumin as blocking agent was alternated with casein in the form of dried milk powder. 1155 IGATAPI(1) SNAYYPH(1) SSLPSVH(1) STPTRFP(1) 4 Phage display (in vivo) 1 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1156 LSTPPLL(2) MHLGDAQ(1) SGPRTLP(1) STYPRHM(1) 4 Phage display (in vivo) 2 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1157 KPSGLTY(1) LSTPPLL(1) SQMPARL(1) YPANLYP(1) YPTSKQS(1) 5 Phage display (in vivo) 3 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1158 APVVRFI(1) KPSGLTY(1) KVLPFYD(1) LSTPPLL(1) QAISRNA(1) 5 Phage display (in vivo) 4 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1159 KVLPFYD(3) LPLTPLP(2) SEAGSRY(1) SQMPARL(1) YPANLYP(1) 5 Phage display (in vivo) 5 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1160 EPPHRHA(1) SAIAQPR(1) SHGPQGY(1) SSFNGLH(1) 4 Phage display (in vivo) 1 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1161 HAIYPRH(4) NPLPLTS(1) 2 Phage display (in vivo) 2 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1162 HAIYPRH(2) GPPSPRY(1) KLTLPNR(1) LSTPPLL(1) 4 Phage display (in vivo) 3 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1163 HAIYPRH(2) KQTLPSA(1) SLPLFTR(1) TVHPPFS(1) 4 Phage display (in vivo) 4 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1164 HAIYPRH(4) ASSDLHS(1) KQTLPSA(1) YNHQRPP(1) YTGPYQH(1) 5 Phage display (in vivo) 5 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1165 KLPPPTF(1) LRFTATI(1) LSQTSGP(1) TVMSQRH(1) 4 Phage display (in vivo) 1 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1166 GLQNSLP(1) LMPSPRN(1) SQMPARL(1) SRTPIIH(1) VPSAPML(1) 5 Phage display (in vivo) 2 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1167 ASYTQPA(1) LPLTPLP(1) SHTTVNP(1) SQMPARL(1) TRTAEGH(1) 5 Phage display (in vivo) 3 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1168 ATFTHYK(1) FFPQPWA(1) KPSGLTY(1) SALPNLP(1) 4 Phage display (in vivo) 4 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1169 KPSGLTY(4) ASYTQPA(2) KQTLPSA(1) SPSAHPS(1) VRRQPAH(1) 5 Phage display (in vivo) 5 PMID:12852441 Polarized human umbilical vein endothelial (HUVE) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phage library, diluted in PBS, were injected into the umbilical vein through the catheter. 1170 ARARS[7659] ARASE[2158] ERVSP[99] ETKRS[4358] GTLRF[5020] GVFRS[3545] IMSRQ[2512] KLRTT[9646] KTRSN[1917] LRMPT[5158] LRSRA[16926] LTTSK[1705] MTRSN[1772] PGRAP[3282] PSARM[5389] RGRMA[5765] RGRSE[14605] RNDKL[1196] RRSID[14982] SGRLA[3042] TGSRD[1417] TLQRV[3436] TRAPM[3299] TRDSR[2548] TVRMP[2289] VLRSP[2756] VRPLE[1248] GTVRS[NT] HRMSS[NT] ISPRS[NT] LGRSL[NT] MRNRA[NT] QWRNS[NT] RAAMM[NT] RLERV[NT] RPQEL[NT] RSGSV[NT] RVGPY[NT] SDRTA[NT] SSPRV[NT] VESRA[NT] 41 Phage display (common panning) 5-6, 8 PMID:12047384 Kallikrein-2 Not determined. Not determined. Not determined. X5 phage display library Fluorescence The CFP fluorescent substrate assay has been utilized to determine the kinetics of peptide substrate selection from a phage display library. Specificity constants (kcat/Km, 1/M × 1/s) were determined and shown. NT denotes not tested. The scissile bonds identified by phage display substrate selection correspond to those of the natural biological substrates of hK2, which include protein C inhibitor, semenogelins, and fibronectin. Moreover, three new putative hK2 protein substrates, shown elsewhere to be involved in the biology of the cancer, have been identified thus reinforcing the importance of hK2 in prostate cancer development. 1171 CPWYFWPC(4)[11.14] CPWYLAPC(2)[14.00] CPWYFLWC(2)[8.56] CWANGWPC(1)[20.26] CHGPLVWC(1)[14.27] P-W-Y-[FL]-W-P 5 Phage display (competitive panning) 3 PMID:11745373 C-C chemokine receptor type 3 Eotaxin Not determined. Not determined. CX6C phage display library Binding assay Results are expressed as percentage (mean ± SD) of (125)I-eotaxin binding. Experiments were performed in duplicate and repeated three times. Data shown were reproduced from the data. In fact, CCR3-transfected murine pre-Bcells were the target. Bound phage were first specifically eluted by competition with excess of eotaxin. 1172 MTRSIA(7)[17.71] LTDYQM(1)[17.16] VTPRQR(1)[11.95] LSSRKI(1)[15.35] 4 Phage display (common panning) 3 PMID:11745373 Eotaxin C-C chemokine receptor type 3 Not determined. Not determined. X6 phage display library Binding assay Results are expressed as percentage (mean ± SD) of (125)I-eotaxin binding. Experiments were performed in duplicate and repeated three times. Data shown were reproduced from the data. 1173 TFAPRWTQSWLP GRLAQYLHHPPT ARMTQLPFAALW GRWMHQPLPQLL GRMTQTSTLLLA GRMMTSIVPLPQ ARMTQTAAFLYT GRLTQTLGYQAQ ARWTQGLPAWGH GRLQPPSILNLW GVGRSLLFRLR SNMHFWNRFFEL DRTPWPRWLLSY 13 Phage display (common panning) 4 PMID:11330351 Botulinum neurotoxin A light chain Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1174 AARHPTMHPSSS RLKKELRLPLAP NKWPLAHSQKKR SLTARQPPRVSW QPTLMTGATMHG FHRHHHSQHLPR HLKHFHHHSPNY KNLHHHHPHRSS KTMHRHHMHWSP HFKHHKSLPHLG 10 Phage display (common panning) 4 PMID:11330351 Botulinum neurotoxin B light chain Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1175 RLKKELRLPLAP KSPKHSSSALHF KSRFRALGASLS LPTQNATVWPQR WHRLRGRITKSG RTRFPYGLRPVR TKRRPRTSPLGL 7 Phage display (common panning) 4 PMID:11330351 Botulinum neurotoxin C1 light chain Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1176 LPYSHKFMIHSKPTF(6) PQHYKMPVRPYSIHS(4) SSTRSLTFDFNMLHS(4) MDAQHLVALHDVAFY(4) SCCTHSTPALPQLPS(4) NAVAHQAVGPAPFLS(3) WTPSTLSHYMTSPFY(2) HLLTHKAHDNAYYAK(2) PTPCHGQVDELHAAV(1) [IL]-H-S 9 Phage display (common panning) 3 PMID:11245205 Cellulose-binding domain (CBD) Not determined. Not determined. Not determined. X15 M13 phage display library The sequence I/LHS, which was found to be an efficient binder of CBD, was fused to a synthetic gene of HRP as an affinity tag. The tagged enzyme (tHRP) was then immobilized on microcrystalline cellulose coated with CBD, thereby demonstrating the indirect immobilization of a protein to cellulose via three amino acids selected by phage display library and CBD. 1177 KLWVIPQ(3) KLWVLPK(2) KLWQVFP(1) KVWILTP(1) KVWTIPR(1) KVWYITP(1) KCCYIPT(4) KGPPITR(1) KVWDLRS(1) YVTREPR(1) K-B-W-x-B-[PTRF]-x 10 Phage display (subtractive panning) 3 PMID:12549820 DNA topoisomerase 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Before applying phage display library to htop(1-214) prepanning step was performed with albumin (100µg/ml) as a target. The consensus sequence identified for clones binding to the N-terminal domain was found in 35 human proteins that are either permanently or temporarily located in the nucleus. They are in majority involved in the DNA repair, transcription, RNA metabolism or cell cycle control. Four of identified proteins: Bub3 protein, Cockayne syndrome protein A, damaged DNA binding protein 2 and GRWD protein belong toWD-repeat proteins and their sequences recognized by the N-terminal domain are identically localized. 1178 LPPSIPR(11) NLRPSIP(9) LLPSIPF(7) LIPSITW(3) L-x-P-S-I-P 4 Phage display (subtractive panning) 3 PMID:12474050 Anti-JL1 leukemia-specific monoclonal antibody JL1 antigen Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage library was pre-incubated with an irrelevant mAb (IgG1, kappa) for 30 min and then added into the well coated with anti-JL1 mAb. 1179 LPPSIPFGLTVG(16)[5.63] LLPSIPNQAYLG(14)[6.50] L-x-P-S-I-P 2 Phage display (subtractive panning) 3 PMID:12474050 Anti-JL1 leukemia-specific monoclonal antibody JL1 antigen Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR) The interaction between anti-JL1 mAb and the mimetic peptides was tested using a surface plasmon resonance biosensor. The Kd (μM) value was measured and shown. The phage library was pre-incubated with an irrelevant mAb (IgG1, kappa) for 30 min and then added into the well coated with anti-JL1 mAb. 1180 CHQKPWEC(17) 1 Phage display (subtractive panning) 2 PMID:12496764 Purified IgGs from serum of prostate cancer patient 1 Not determined. Not determined. Not determined. CX6C phage display library First, a pre-clearing step was used to remove nonspecific clones by pre-absorbing the phage peptide library onto purified IgGs from normal serum pooled from volunteer agematched blood-donor control men. Next, the pre-cleared phage library was selected onto the pool of IgGs purified from the serum of prostate cancer patients. Authors identified a consensus motif, NX[ST]DK[ST], that bound selectively to circulating antibodies from cancer patients over control antibodies from blood donors. Authors validated this motif by showing that positive serum reactivity to the peptide was specifically linked to disease progression and to shorter survival in a large patient population. 1181 CHQKPWEC(14) 1 Phage display (subtractive panning) 3 PMID:12496764 Purified IgGs from serum of prostate cancer patient 1 Not determined. Not determined. Not determined. CX6C phage display library First, a pre-clearing step was used to remove nonspecific clones by pre-absorbing the phage peptide library onto purified IgGs from normal serum pooled from volunteer agematched blood-donor control men. Next, the pre-cleared phage library was selected onto the pool of IgGs purified from the serum of prostate cancer patients. Authors identified a consensus motif, NX[ST]DK[ST], that bound selectively to circulating antibodies from cancer patients over control antibodies from blood donors. Authors validated this motif by showing that positive serum reactivity to the peptide was specifically linked to disease progression and to shorter survival in a large patient population. 1182 CKDRFERC(17) 1 Phage display (subtractive panning) 2 PMID:12496764 Purified IgGs from serum of prostate cancer patient 2 Not determined. Not determined. Not determined. CX6C phage display library First, a pre-clearing step was used to remove nonspecific clones by pre-absorbing the phage peptide library onto purified IgGs from normal serum pooled from volunteer agematched blood-donor control men. Next, the pre-cleared phage library was selected onto the pool of IgGs purified from the serum of prostate cancer patients. Authors identified a consensus motif, NX[ST]DK[ST], that bound selectively to circulating antibodies from cancer patients over control antibodies from blood donors. Authors validated this motif by showing that positive serum reactivity to the peptide was specifically linked to disease progression and to shorter survival in a large patient population. 1183 CKDRFERC(14) 1 Phage display (subtractive panning) 3 PMID:12496764 Purified IgGs from serum of prostate cancer patient 2 Not determined. Not determined. Not determined. CX6C phage display library First, a pre-clearing step was used to remove nonspecific clones by pre-absorbing the phage peptide library onto purified IgGs from normal serum pooled from volunteer agematched blood-donor control men. Next, the pre-cleared phage library was selected onto the pool of IgGs purified from the serum of prostate cancer patients. Authors identified a consensus motif, NX[ST]DK[ST], that bound selectively to circulating antibodies from cancer patients over control antibodies from blood donors. Authors validated this motif by showing that positive serum reactivity to the peptide was specifically linked to disease progression and to shorter survival in a large patient population. 1184 CNWTDKTC(4) CNITQKSC(4) CNVSDKSC(2) N-x-[ST]-D-K-[ST] 3 Phage display (subtractive panning) 2 PMID:12496764 Purified IgGs from serum of prostate cancer patient 3 Not determined. Not determined. Not determined. CX6C phage display library First, a pre-clearing step was used to remove nonspecific clones by pre-absorbing the phage peptide library onto purified IgGs from normal serum pooled from volunteer agematched blood-donor control men. Next, the pre-cleared phage library was selected onto the pool of IgGs purified from the serum of prostate cancer patients. Authors identified a consensus motif, NX[ST]DK[ST], that bound selectively to circulating antibodies from cancer patients over control antibodies from blood donors. Authors validated this motif by showing that positive serum reactivity to the peptide was specifically linked to disease progression and to shorter survival in a large patient population. 1185 CNVSDKSC(13) CNKTDKGC(2) CNWTDKTC(1) N-x-[ST]-D-K-[ST] 3 Phage display (subtractive panning) 3 PMID:12496764 Purified IgGs from serum of prostate cancer patient 3 Not determined. Not determined. Not determined. CX6C phage display library First, a pre-clearing step was used to remove nonspecific clones by pre-absorbing the phage peptide library onto purified IgGs from normal serum pooled from volunteer agematched blood-donor control men. Next, the pre-cleared phage library was selected onto the pool of IgGs purified from the serum of prostate cancer patients. Authors identified a consensus motif, NX[ST]DK[ST], that bound selectively to circulating antibodies from cancer patients over control antibodies from blood donors. Authors validated this motif by showing that positive serum reactivity to the peptide was specifically linked to disease progression and to shorter survival in a large patient population. 1186 STVPRMPLSPPN FMNPLHPRVLRP NYVIHDVPRHPA VNMGNYRQQQPL SMLDLFPRAASY SVYNVRPSSLSA TVMASPTMKSNS VNMGNYRQQQPQ KFPGPNCCHALP 9 Phage display (common panning) 5 PMID:12409405 Mycobacterium paratuberculosis Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1187 SLRLLQWFLWAC(6) 1 Phage display (common panning) 3 PMID:11318108 Deoxyribonuclease-2 Not determined. Not determined. Not determined. X12 phage display library One conserved sequence (SLRLLQWFLWAC) was found in six colonies, while all other sequences appeared only once. 1188 RLQAWWEMWADSYRP(5)[0.80 ± 0.03] QKRPVNISTLQELWE(3)[0.34 ± 0.02] SVQARWEAAFDLDLY(3)[0.52 ± 0.09] [LV]-Q-[AE]-x-W-E 3 Phage display (common panning) 3 PMID:12542398 Anti-human IFN-b monoclonal antibody YSB-2 Interferon beta, IFN-beta Not determined. Not determined. f3-15mer phage display library (X15) ELISA Absorbances at 450 nm was measured. Data, means ± S.D. from triplicate wells, were reproduced from the graph. Authors screened a random phage display library with a soluble form of the human type I IFN receptor (sIFNR) and a neutralizing monoclonal antibody (mAb) against IFN-b. Three clones were identified as binding YSB-2 ,but screening with sIFNR failed to identify any peptide ligands. 1189 YAPYGL(2)[88/82] WLPSSI(1)[96] RLPLGI(1)[94] QRPHAI(1)[93] KGPIHL(1)[91] FLPQGW(1)[90] LDPWSY(1)[81] LLGLRN(1)[91] QLLGFL(1)[90] AVVQHL(1)[89] NALASI(1)[88] ARLGHF(1)[85] GAVKHL(1)[81] GAARHI(1)[79] NLRSKL(1)[79] NLKSRV(1)[77] DRQMAL(1)[69] FSMFSL(1)[66] WAKSNL(1)[48] VSLKNL(1)[45] TVGWIS(1)[33] VASGIY(1)[86] PQHVRQ(1)[71] AVYFHM(1)[66] LRAMQT(1)[65] GANSLK(1)[63] RIGFLR(1)[88] VFSIPL(1)[79] RAMHMY(1)[79] RSENIR(1)[79] IKYHSL(1)[77] LISHSI(1)[73] ARYRWL(1)[72] GVSMPF(1)[67] VMSIRI(1)[61] RSYAIL(1)[55] SRRLHL(1)[50] FIANPV(1)[42] 38 Phage display (common panning) 3 PMID:11959855 Matrix metalloproteinase-14, MMP-14 Not determined. Not determined. Not determined. X6 phage display library ELISA Individual phage were analyzed after three rounds of selection by MT1-MMP. The substrates are grouped according to their ability to be hydrolyzed by MT1-MMP, relative to non-treated controls, as measured by substrate phage ELISA and expressed as % hydrolysis. Two-hundred clones were screened for their ability to be hydrolyzed by MT1-MMP using the substrate phage ELISA we reported previously. Forty of the two-hundred clones were cleaved efficiently by MT1-MMP and were carried forward for additional analysis. 1190 CPEFVDVEGDPGALLAC CGWVDVIASGDTATLAC CDVEWVDVSSLEWDLPC CLMGCWCDVGVGGESLC CPDWVDVFKLVEGVMLC [ED]-[WF]-[VC]-D-V 5 Phage display (common panning) 5-7 PMID:12393589 P-selectin P-selectin glycoprotein ligand 1 1G1S, Not determined. pComb8 CX15C phage display library In fact, chimeric P-selectin, consisting of human IgG1 fused to the binding domain of human P-selectin (PS-IgG), was the target. 1191 VGLDPRDWVDVSDYA DWVDVREVLTGEQRV DWVDV 2 Phage display (common panning) PMID:12393589 P-selectin P-selectin glycoprotein ligand 1 1G1S, Not determined. pIF4 X15 phage display library In fact, chimeric P-selectin, consisting of human IgG1 fused to the binding domain of human P-selectin (PS-IgG), was the target. 1192 ECSEGWCDMKIDRLDAGG WCDSVGDRAGPSV WCD 2 Phage display (common panning) PMID:12393589 P-selectin P-selectin glycoprotein ligand 1 1G1S, Not determined. pIF4 X28 phage display library In fact, chimeric P-selectin, consisting of human IgG1 fused to the binding domain of human P-selectin (PS-IgG), was the target. The isolated peptide sequences were not displayed completely. 1193 FLHTRLFVSDWYHTP(2) SRAPRFPWLNPLDGW(2) TFTRVVTDVYRPRLS(2) EDFRFESSLSINDHA(2) GWHTRIFVSDWYHTP(1) SLAPRFPWLNPLDGW(1) SRSPRFPWLNPLDGW(1) TFMRVVTDVYRPRLS(1) SFTRVVTDVYRPRLS(1) DFRFESSLSINDHA(1) 10 Phage display (subtractive panning) 3 PMID:12472892 Anti-L2/HNK-1 monoclonal antibody L2-412 L2/HNK-1 carbohydrate Not determined. Not determined. f3-15mer phage display library (X15) For the second and third rounds of screening, the phages were preincubated with rat IgG (100 μg/mL) to decrease non-specific binding. The phages isolated from a 15-mer peptide library by adsorption to this antibody share a consensus sequence of amino acids. The peptide mimicked several important functions of the L2/HNK-1 carbohydrate, such as binding to motor neurons in vitro, and preferential promotion of in vitro neurite outgrowth from motor axons compared with sensory neurons. 1194 VTWRMWYVPA(2) VVHMRYSFRI(5) YEIRFRYLQF(4) TGYRLVIYRL(1) ASTIRISFFV(3) AIRIVIATNI(3) SEGAVVFHAT(2) KIQLQWIVSL(2) YSGSLVLIVR(2) SHSLKLTLVI(4) MMKVVFRWSD(1) TLGGGLLLWH(1) VPVLHWTVMF(1) RVVYISAMVT(1) LVVINITYDR(1) AWTSPVDSFQ(1) YLHIVFQVPT(1) PRGYWLRIEW(1) RMRY 18 Phage display (common panning) 3 PMID:9531425 Cholesteryl ester transfer protein Cholesteryl esters, CE Not determined. Not determined. X10 phage display library Of the 36 randomly chosen clones, phage displaying VTWRMWYVPA, VVHMRYSFRI, YEIRFRYLQF and TGYRLVIYRL exhibited relatively high binding to CETP (OD450 0.4-0.7), whereas the remaining 24 clones displayed only moderate to low binding to CETP in the reported assay system. The synthesized VTWRMWYVPA inhibited CETP-catalyzed transfer of cholesteryl esters and triglycerides. Amino- and ciirboxy-terminal truncations demonstrated that the original decapeptide could be reduced to a pentapeptide without loss of either its ability to bind to CETP or its ability to inhibit CETP-mediated lipid transfer. That pentapeptide, WRMWY bound directly to CETP and its inhibition was consistent with that of a competitive inhibitor of CETP. 1195 SRPKPPNPS(15)[NT] KRGSIIPNG(5)[1.225, 1.267, 2.066] KTKKPPNPS(4)[NT] RLKPPNPTE(2)[NT] KPKTNQIRP(2)[NT] YTTTLSFQK(2)[NT] GVHRLTRPS(1)[1.035, 5.124, 0.888] GLHRSQTSM(1)[0.904, 4.587, 1.031] GRHHWASGP(1)[0.933, 5.390, 1.314] GQHKSRSFP(1)[0.976, 7.768, 0.664] RKPPNPPPP(1)[NT] KPPNPTRPE(1)[NT] KPPNPRPSL(1)[NT] RPSKPPNPV(1)[NT] KALKPPNPV(1)[NT] RRDTISPYS(1)[NT] KKTGNITPK(1)[NT] GQHHSLSTP(1)[NT] GAHASLWTP(1)[NT] GSHLSARAP(1)[NT] GMHKSVFAT(1)[NT] DTTLNYRGA(1)[NT] 22 Phage display (common panning) 1 PMID:9532590 IgG from the cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients Not determined. Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA Binding of the selected phagotopes to the CSF1, CSF2 and CSF3 was detected by ELISA on immobilized phage, respectively. For each sample antibody recognition of the tested phagotope and wild type phage was measured. Average values (A405nm) from two independent experiments have been determined. Results are expressed as the ratio between the average value of the tested phagotope and that of wild type phage (clone/wt A405 nm). Data shown were reproduced from the graph. NT denotes not tested. For each selection, 5 microgram of CSF IgG were immobilized using magnetic beads coated with goat anti-human IgG Fc-specific Ab. Eluted phages were subjected to immunological screening using the same CSF as probes. Positive plaques were individually picked and rescreened using CSF from MS patients and from patients affected by other neurological diseases. Fifty-seven phages positive only to the MS CSF were identified. Their sequences were mixed together in the original paper, though the two libraries were selected separately. 1196 WLQDITL(1) IDQDITV(1) QQDITVH(1) QDITLRP(1) HIRRILL(2) YPMIISP(1) ENGADVD(1) VHPPPFL(1) QDIT 8 Phage display (competitive panning) 3 PMID:9598980 Anti-syndecan-1 monoclonal antibody B-B2 Syndecan-1, SYND1 Not determined. Not determined. Ph.D.-7 phage display library (X7) B-B2 was produced by ascitic fluid and purified on a protein A column. Coated mAb was used to select the phages which were then eluted with an acidic treatment, and amplified in bacteria for the next round of selection. After the third round of selection bound phages were eluted by adding the selecting mAb as competitor. 1197 IQDITLS(1) RQDITLV(1) GHTQDIT(1) QDITLFN(4) QDITMFV(2) MQNITLF(1) QNITLSQ(1) QDIT 7 Phage display (common panning) 3 PMID:9598980 Anti-syndecan-1 monoclonal antibody B-B2 Syndecan-1, SYND1 Not determined. Not determined. Ph.D.-7 phage display library (X7) B-B2 was produced by ascitic fluid and purified on a protein A column. Coated mAb was used to select the phages which were then eluted with an acidic treatment, and amplified in bacteria for the next round of selection. After the third round of selection bound phages were eluted by adding the selecting mAb as competitor and then acidic conditions were applied to recover the remaining phages. Independently of the elution procedure used, the number of recovered phages was at least 30 times higher from the well coated with the selecting mAb than with an irrelevant control mAb, suggestting that the selected phages were mAb specific. 1198 LTLPEVA(1) APLPEVL(1) HLPEVSP(1) FLPEVLL(1) YLPEVIS(1) YLPEVLP(1) YLPEVQP(2) FLPDVAS(1) FLPDVLS(1) YLPQVAS(1) LTDVRTP(1) TTNDFTR(1) LPEV 12 Phage display (competitive panning) 3 PMID:9598980 Anti-syndecan-1 monoclonal antibody B-B4 Syndecan-1, SYND1 Not determined. Not determined. Ph.D.-7 phage display library (X7) B-B4 was produced by ascitic fluid and purified on a protein A column. Coated mAb was used to select the phages which were then eluted with an acidic treatment, and amplified in bacteria for the next round of selection. After the third round of selection bound phages were eluted by adding the selecting mAb as competitor. 1199 ILPEVVS(1) TLPEVLS(1) QLPEVSS(1) ALPEVLS(1) MLPEVLP(1) LELPEVM(1) VLQEVSS(1) AFPSPTL(1) AYTSAHK(1) LPEV 9 Phage display (common panning) 3 PMID:9598980 Anti-syndecan-1 monoclonal antibody B-B4 Syndecan-1, SYND1 Not determined. Not determined. Ph.D.-7 phage display library (X7) B-B4 was produced by ascitic fluid and purified on a protein A column. Coated mAb was used to select the phages which were then eluted with an acidic treatment, and amplified in bacteria for the next round of selection. After the third round of selection bound phages were eluted by adding the selecting mAb as competitor and then acidic conditions were applied to recover the remaining phages. Independently of the elution procedure used, the number of recovered phages was at least 30 times higher from the well coated with the selecting mAb than with an irrelevant control mAb, suggestting that the selected phages were mAb specific. 1200 WWIWWPGSVE(4) WWGPVIMWDA(2) WFSWGFPQWW(1) VWWSLLVPLA(4) SWGFPVFWHR(1) STGFPIFWHS(1) PEGEYAWWVE(1) DWWSYAWTLV(1) WLWVSRWPGG(2) WWLLWTIPSW(1) GMWWWPLFTL(1) PQVWGVWTDE(1) VLNWPGMLLV(1) WHLACAGVFS(1) WWTLIYLMP(1) 15 Phage display (common panning) 3 PMID:9661811 MDDQRDLISNNEQLK peptide Not determined. Not determined. Not determined. fUSE5-based X10 phage display library When screening a large number of individual phage clones for binding to biotinylated Ii peptide (MDDQRDLISNNEQLK) coupled to microtiter wells, seven phage clones binding significantly above background level were identified. Sequencing of phage inserts revealed that one displayed WFSWGFPQWW, two had insert WWGPVIMWDA, and four contained WWIWWPGSVE. 1201 CPALLCRC(1) CIPALWSFC(2) CAARLWQWC(1) CTA(A/P)LWRIC(1) CTAALWRIC(2) CTATLWRIC(2) 6 Phage display (common panning) 6 PMID:9680646 Chymotrypsinogen A Not determined. Not determined. Not determined. CX7C phage display library Phage particles were incubated with biotinylated alpha-chymotrypsin in pH 6.5 buffer. Streptavidin-agarose (50 μl) was blocked with 20 μl 50 mg/ml dialyzed BSA in TBS for 60 min and added to each of the three selection vials. 1202 CSASLWFLC(13) CS(G/E)TLWS(K/T)C(1) 2 Phage display (common panning) 6 PMID:9680646 Chymotrypsinogen A Not determined. Not determined. Not determined. CX7C phage display library Phage particles were incubated with biotinylated alpha-chymotrypsin in pH 7.0 buffer. Streptavidin-agarose (50 μl) was blocked with 20 μl 50 mg/ml dialyzed BSA in TBS for 60 min and added to each of the three selection vials. 1203 CCFSWRCRC(5) CWGSWRCRC(1) CWSPKRCRC(1) 3 Phage display (common panning) 6 PMID:9680646 Chymotrypsinogen A Not determined. Not determined. Not determined. CX7C phage display library Phage particles were incubated with biotinylated alpha-chymotrypsin in pH 7.5 buffer. Streptavidin-agarose (50 μl) was blocked with 20 μl 50 mg/ml dialyzed BSA in TBS for 60 min and added to each of the three selection vials. The oxidized form of the synthetic peptide CCFSWRCRC, selected at pH 7.5, could completely inhibit the enzymatic activity of alpha-chymotrypsin. The structurally related enzymes trypsin (bovine) and elastase (porcine) were only marginally inhibited by the same peptide under the same conditions. 1204 RRLPFGSQM(4)[+++] RYAFGSQIA(2)[++] RRLPFGSSL(1)[+] RAGRFGYQR(1)[+] TRSFGIQAT(1)[+] SRLAFGDQL(1)[+] HEHTFGRQW(1)[+++] TRAFGNEAT(1)[+] RAAPFGNQW(1)[++] HRLAFGQNT(1)[+++] HRLAFGQYT(1)[+++] F-G-x-Q 11 Phage display (subtractive panning) 3 PMID:9693968 Anti-collagen alpha-1(II) chain monoclonal antibody CII-C1 Collagen alpha-1(II) chain Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA Phagotopes selected by biopanning with the mAbs were tested by ELISA for reactivity with CII-C1. The negative control was wild type fl bacteriophage (wt fl). After the first round of positive selection, there was a negative selection step in which phage not specifically reactive with the monoclonal antibody used for selection were removed using magnetic beads coated with anti-mouse immunoglobulin. 1205 CIAPKRHNSAC[++] CESAQRPFGCC[++] 2 Phage display (subtractive panning) 3 PMID:9693968 Anti-collagen alpha-1(II) chain monoclonal antibody CII-C1 Collagen alpha-1(II) chain Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA Phagotopes selected by biopanning with the mAbs were tested by ELISA for reactivity with CII-C1. The negative control was wild type fl bacteriophage (wt fl). After the first round of positive selection, there was a negative selection step in which phage not specifically reactive with the monoclonal antibody used for selection were removed using magnetic beads coated with anti-mouse immunoglobulin. 1206 LSFAAEGEF(1)[NT] YKLTCSSAQ(1)[NT] ARLSSAVVK(1)[-] GHSLCSCIP(1)[-] GSRFNKVPS(1)[-] KKKTVLPDL(1)[-] LKIGDFPAG(1)[-] SRRTLMSGA(1)[-] TKIRNVAGM(1)[-] 9 Phage display (subtractive panning) 3 PMID:9693968 Anti-chicken GAD monoclonal antibody GAD1 Glutamate decarboxylase 67 Not determined. Not determined. pVIII-9aa phage display library (X9) Phagotopes selected by biopanning with the mAbs were tested by ELISA for reactivity with GAD1. The negative control was wild type fl bacteriophage (wt fl). NT represents not tested. After the first round of positive selection, there was a negative selection step in which phage not specifically reactive with the monoclonal antibody used for selection were removed using magnetic beads coated with anti-mouse immunoglobulin. 1207 EPIHRSTLTALL(1) EPIHRSTLTAPL(1) YQDMIYMRPLDS(1) PRPSPKMGVSV(1) ASNHTHSSSIQF(3) TATHRHSSSI(1) DARHSSSLQMLF(1) FSLRPTMNFTNL(1) DPGKIYFHIAVS(1) 9 Phage display (common panning) 4 PMID:10076823 Constant region of humanized anti-Tac monoclonal antibody (HAT) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The pFc' fragments of a humanized IgG1 monoclonal antibody were generated by digestion with immobilized pepsin. These pFc' fragments were separated from F(ab')2 fragments by affinity chromatography. The pFc' fragments corresponding to the constant region of the humanized anti-Tac (HAT) IgG1 monoclonal antibody were used as targets for panning. EPIHRSTLTALL was the best peptide, which had a binding capacity of 320 mg HAT/g gel in affinity chromatography of HAT. 1208 RQLVQPL(1) SPAPSDS(1) SSQSDPA(1) SKPTQLH(1) GTATSPH(1) SHLGFDD(3) TSDTGWR(1) 7 Phage display (common panning) 4 PMID:10076823 Constant region of humanized anti-Tac monoclonal antibody (HAT) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The pFc' fragments of a humanized IgG1 monoclonal antibody were generated by digestion with immobilized pepsin. These pFc' fragments were separated from F(ab')2 fragments by affinity chromatography. The pFc' fragments corresponding to the constant region of the humanized anti-Tac (HAT) IgG1 monoclonal antibody were used as targets for panning. 1209 RVRPAP(5) LERLPP(2) VRLPPN(9) NIRLPP(7) KIRLPP(1) ERRAPG(2) EIRRAP(1) VRYPPR(1) KSKAGV(1) R-x(2)-P 9 Phage display (common panning) 3 PMID:16695994 Anti-mucin monoclonal antibody C595 Mucin-1, MUC-1 Not determined. Not determined. f3-6mer phage display library (X6) 1210 GERWCFDGPLTWVCGEES 1 Phage display (common panning) 4 PMID:9922141 Vascular endothelial growth factor A, VEGF-A Vascular endothelial growth factor receptor 2, VEGFR-2 Not determined. Not determined. X4CX2GPX4CX4 phage display library The receptor binding domain of human VEGF (residues 8-109) was overexpressed in Escherichia coli inclusion bodies, purified, refolded and used as the target for panning. Sequencing of representitive clones yielded a single consensus sequence. Phage ELISA experiments showed that the binding of these phage clones to VEGF could be blocked by KDR. 1211 RGWVEICVADDNGMCVTEAQ GWDECDVARMWEWECFAGV 2 Phage display (common panning) 4 PMID:9922141 Vascular endothelial growth factor A, VEGF-A Vascular endothelial growth factor receptor 2, VEGFR-2 Not determined. 1IDH,1VPP, X(i)CX(j)CX(k) phage display library pool The receptor binding domain of human VEGF (residues 8-109) was overexpressed in Escherichia coli inclusion bodies, purified, refolded and used as the targetfor panning. Sequencing of representitive clones yielded two predominant sequences. Phage ELISA experiments showed that the binding of these phage clones to VEGF could be blocked by KDR. 1212 SYSWMYE(1400)[-, 0.69] YWDGQW(22)[-, 0.20] WWDGQF(13)[-, 0.17] YDWLYE(6)[-, 0.42] DYAWMY(2)[-, 0.67] 5 Phage display (common panning) 4 PMID:8760499 Anti-GXM monoclonal antibody 2H1 Glucuronoxylomannan, GXM Not determined. Not determined. f3-6mer phage display library (X6) ELISA Large-scale phage preparations were made using PEG precipitation. The direct ELISA was done by absorbing anti-phage antibody to the ELISA plate and then adding 2e10 phage. 2H1 (2 μg/ml) was then added and the ELISA was developed with an alkaline phosphatase-conjugated anti-γ1 antibody. The enhanced direct ELISA was carried out the same way with a threefold excess of anti-γ1 antibody (6 μg/ml) added together with 2H1. For the hexapeptide library, complexes between 2H1 and phage were captured with streptavidin-coated magnetic beads (Advanced Magnetics, Inc., MA). 1213 LQYTPSWMLV(18)[0.51, >2.0] NSPETPDWMV(2)[-, 0.65 ±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hage display (common panning) 4 PMID:8760499 Anti-GXM monoclonal antibody 2H1 Glucuronoxylomannan, GXM Not determined. 2H1P, L100 phage display library (X10) ELISA Large-scale phage preparations were made using PEG precipitation. The direct ELISA was done by absorbing anti-phage antibody to the ELISA plate and then adding 2e10 phage. 2H1 (2 μg/ml) was then added and the ELISA was developed with an alkaline phosphatase-conjugated anti-γ1 antibody. The enhanced direct ELISA was carried out the same way with a threefold excess of anti-γ1 antibody (6 μg/ml) added together with 2H1. For the decapeptide library, complexes between 2H1 and phage were captured with streptavidin-coated magnetic beads (Advanced Magnetics, Inc., MA) and a biotinylated anti-mouse IgG1 antibody. LQYTPSWMLV binds to 2H1 with a Kd of 295nM. The structure of GLQYTPSWMLVG complexed with 2H1 has been solved (PDB ID: 2H1P). 1214 AGRDRG[0.652] AGRDLL[1.572] RMKSSR[0.378] RLKSSR[0.988] VKKSGR[0.203] HLFEAP[0.225] HLFEAH[1.539] KQSRRG[0.379] TLDTAH[0.612] RLRVKN[0.296] DTGHHR[0.430] HRGRAQ[0.291] HRGRAA[0.343] KPDSMP[0.424] FTILRY[0.734] FFEVAH[0.513] AGRDRA[NT] AGRDRF[NT] AGRDLF[NT] AGRDSH[NT] AGRDSV[NT] RLKSMR[NT] RFKSMR[NT] VFKSAR[NT] VRKSAR[NT] IRKSSR[NT] KTSRRG[NT] KLARRG[NT] GLVEAH[NT] VLYQAH[NT] HLYSAH[NT] KLWMAH[NT] RDQHNS[NT] RDQHKS[NT] KAQEKV[NT] RAQEKV[NT] LAQDKV[NT] LAQHKS[NT] RGRLDR[NT] HHRGSA[NT] WTKQYR[NT] KNTYPI[NT] RQYRPG[NT] SQMPIF[NT] 44 Phage display (common panning) 3-4 PMID:10921846 IgG fraction of anti-EHV-1 serum Equine herpesvirus type 1 Not determined. Not determined. f3-6mer phage display library (X6) ELISA ELISA measurements of the binding to sera from EHV-1 infected gnotobiotic foals expressed as A490. The samples were taken in duplicate and the results are expressed as an average value. NT denotes not tested. Briefly phage (3e9-9e10 plaque forming units, pfu) in 100 ml PBSTG (phosphate buffered saline containing 0.05% Tween 20 and 0.5% goat sera, Seralabs) were incubated with or without biotinylated equine IgG for 5h at 37°C. An equal volume of M-280 streptavidin-coated magnetic beads (Dynal), washed three times with PBSTG, was added, mixed and stored at room temperature for 5 min. The sequences of the amino acids displayed were aligned with protein sequences of EHV-1, thereby identifying a number of potential antibody binding regions. Presumptive epitopes were identified within the proteins encoded by genes 7 (DNA helicase:primase complex protein), 11 (tegument protein), 16 (glycoprotein C), 41 (integral membrane protein), 70 (glycoprotein G), 71 (envelope glycoprotein gp300), and 74 (glycoprotein E). Two groups of sequences, which aligned with either glycoprotein C (gC) or glycoprotein E (gE), identified type-specific epitopes which could be used to distinguish between sera from horses infected with either EHV-1 or EHV-4 in an ELISA using either the phage displaying the peptide or synthetic peptides as antigen. The gC epitope had been previously identified as an immunogenic region by conventional monoclonal antibody screening whereas the gE antibody binding region had not been previously identified. 1215 KSKRGHLLL[2.870] HHRGAAPNS[2.973] PIRVPGFLG[0.110] PRLKHVQSV[0.227] SQLRTRLKG[NT] PPRVPTTLR[NT] IPPALSISQ[NT] FRRTVNNKL[NT] RLNPHHEPM[NT] ATFTKDAAT[NT] 10 Phage display (common panning) 2 PMID:10921846 IgG fraction of anti-EHV-1 serum Equine herpesvirus type 1 Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA ELISA measurements of the binding to sera from EHV-1 infected gnotobiotic foals expressed as A490. The samples were taken in duplicate and the results are expressed as an average value. NT denotes not tested. Briefly phage (3e9-9e10 plaque forming units, pfu) in 100 ml PBSTG (phosphate buffered saline containing 0.05% Tween 20 and 0.5% goat sera, Seralabs) were incubated with or without biotinylated equine IgG for 5h at 37°C. An equal volume of M-280 streptavidin-coated magnetic beads (Dynal), washed three times with PBSTG, was added, mixed and stored at room temperature for 5 min. The sequences of the amino acids displayed were aligned with protein sequences of EHV-1, thereby identifying a number of potential antibody binding regions. Presumptive epitopes were identified within the proteins encoded by genes 7 (DNA helicase:primase complex protein), 11 (tegument protein), 16 (glycoprotein C), 41 (integral membrane protein), 70 (glycoprotein G), 71 (envelope glycoprotein gp300), and 74 (glycoprotein E). Two groups of sequences, which aligned with either glycoprotein C (gC) or glycoprotein E (gE), identified type-specific epitopes which could be used to distinguish between sera from horses infected with either EHV-1 or EHV-4 in an ELISA using either the phage displaying the peptide or synthetic peptides as antigen. The gC epitope had been previously identified as an immunogenic region by conventional monoclonal antibody screening whereas the gE antibody binding region had not been previously identified. 1216 YFHDRLPKPQTMKPS[0.193] TSLSWSKPPTRSSPI[0.724] LIVQDRPPSRKQHPP[0.167] MHKNNQFHWPSIRYL[0.160] MSPILDFPSRKTHKT[1.265] NSQLTKSPSMKHRVE[1.171] SLASRPHHPLKNYAP[0.113] FVAKSPAKHNPVSVR[0.148] TSMARYVRHSNYKHM[0.219] SCNDNKSVPCIVPQL[0.652] QPRLSTKKTPSSPIA[NT] LPDRPRTQPFVHELP[NT] TSLPPPRQPHPHFVP[NT] IIQYRIPHRPPTQHL[NT] KYPTPQHPTKFTQHS[NT] MPFLINRWPPVNQLL[NT] PPPRTPTQSLPMTFT[NT] SEFRHPHPHLPPFDV[NT] WHTFADMQLMYPLKE[NT] WLPKYPLESTNSLTN[NT] AGISRTAYPTRVPYT[NT] 21 Phage display (common panning) 3 PMID:10921846 IgG fraction of anti-EHV-1 serum Equine herpesvirus type 1 Not determined. Not determined. X15 M13 phage display library ELISA ELISA measurements of the binding to sera from EHV-1 infected gnotobiotic foals expressed as A490. The samples were taken in duplicate and the results are expressed as an average value. NT deneotes not tested. Briefly phage (3e9-9e10 plaque forming units, pfu) in 100 ml PBSTG (phosphate buffered saline containing 0.05% Tween 20 and 0.5% goat sera, Seralabs) were incubated with or without biotinylated equine IgG for 5h at 37°C. An equal volume of M-280 streptavidin-coated magnetic beads (Dynal), washed three times with PBSTG, was added, mixed and stored at room temperature for 5 min. The sequences of the amino acids displayed were aligned with protein sequences of EHV-1, thereby identifying a number of potential antibody binding regions. Presumptive epitopes were identified within the proteins encoded by genes 7 (DNA helicase:primase complex protein), 11 (tegument protein), 16 (glycoprotein C), 41 (integral membrane protein), 70 (glycoprotein G), 71 (envelope glycoprotein gp300), and 74 (glycoprotein E). Two groups of sequences, which aligned with either glycoprotein C (gC) or glycoprotein E (gE), identified type-specific epitopes which could be used to distinguish between sera from horses infected with either EHV-1 or EHV-4 in an ELISA using either the phage displaying the peptide or synthetic peptides as antigen. The gC epitope had been previously identified as an immunogenic region by conventional monoclonal antibody screening whereas the gE antibody binding region had not been previously identified. 1217 LPPPSYWSQ DPPSYHASS APPSYHSSV WFNPPPYPG FYWPPPYTA SMLPPPYPLP FLPPAYRKE SPPAYASSR YRPPAYANA RRPPAYPGA RPPPPYHPE NLRPPAYLQ RAPPTYERS IRSPPPYEP RPPPYARAP MRPPPYVAP RRPPAYEGWHNV HRPEPPPYGNHGH VHPPAYSYYGHE P(2)-x-Y 19 Phage display (common panning) PMID:10767429 WW-EF-ZZ fusion protein from utrophin Not determined. Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) Authors found that none of the carboxy-terminal domains of utrophin, when isolated from its structural context, selects specific ligand peptides from a phage-displayed peptide library. By contrast, panning with an extended region containing the WW, EF hands, and ZZ domain defines the consensus binding motif, PPxY which is also found in L-dystroglycan, a component of the DAP complex that interacts with utrophin in several tissues. 1218 FYPSYHSTPQRP AYYKTASLAPAE SLSLLTMPGNAS 3 Phage display (subtractive panning) 4 PMID:15187120 Human dendritic cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phages were incubated serially with monocytes, T cells, and B cells, then with Langerhans-like DC, then MDC. 1219 HWKHPWGAWDTL HHWHHWCMPHKT HWSAWWIRSNQS HNWYHWWMPHNT 4 Phage display (common panning) 5 PMID:12612679 Carbon nanotubes Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Analysis of peptide conformations shows that the binding sequence is flexible and folds into a structure matching the geometry of carbon nanotubes. The hydrophobic structure of the peptide chains suggests that they act as symmetric detergents. 1220 CLYTRYWC 1 Phage display (common panning) 4 PMID:14978303 SEM-5 SH3 domain Not determined. Not determined. Not determined. CX6C FUSE5-based phage display library For library selection, streptavidin magnesphere paramagnetic particles (SA-PMPs; Promega Cat. #Z5481) were rinsed three times with Tris-buffered saline (TBS; 50 mM Tris-HCl, pH 7.5, 0.15 M NaCl). Biotinylated SH3 (100μg) was added to 0.6 mg SA-PMPs in 600 μL TBS and incubated at 4°C for 18 h. The SA-PMPs were then incubated with 10 μmole of biotin at 4°C for 4 h to block unbound sites. The library phage virions (input = e12 cfu) were added and incubated with the beads in 700 μL TBS at 4°C for 18 h. 1221 CAYTAFPLDC(13) CNTAFPSGTSC(7) 2 Phage display (common panning) PMID:12878179 Anti-mouse CD45 monoclonal antibody IBL-8 Receptor-type tyrosine-protein phosphatase C Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) Among these 20 clones we have identified two types of clones displaying slightly different nonapeptides. A comparison of the deduced amino acid sequence of these two groups of clones with the primary sequence of the CD45 molecule assigns the epitope recognised by the IBL-8 mAb to amino acids 136-144 (ADTAFPVDT). 1222 NHFLIKP(6) NHFLRSP(3) NHFLPRW(3) NHFLPKV(8) NHFLLPP(1) NHFLPPR(1) NHFLPTG(1) NHFLMPK(1) NHFLKGT(1) NHFLPQN(1) NHFLLWR(1) NHFLIRK(2) NHFLPTA(1) NHFL 13 Phage display (common panning) 4 PMID:14602648 Bacillus subtilis spores Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 1223 NHFLKSQPGVVT(2) NHFLNRPAQSQV(1) NHFLPPKMGPTD(1) NHFLPEPRLVMP(1) NHFLAPQPPVKP(2) NHFLMPNPLLAM(1) NHFLIPPEPLRE(1) NHFLPLNPPAPS(1) NHFL 8 Phage display (common panning) 4 PMID:14602648 Bacillus subtilis spores Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1224 CGVVRGPSRGC(5) CALARAARLGC(1) 2 Phage display (subtractive panning) 3 PMID:12210982 Glutathione S-transferase Mu 2 Not determined. Not determined. Not determined. CX9C phage display library The library was mixed together and added to a well preblocked with nonfat dried milk in PBS. And the library was incubated for 1 hour at RT prior to transfer to GST-M2-2-coated microtiter wells. 1225 CAENRFDADLRSSALAC(1) 1 Phage display (subtractive panning) 3 PMID:12210982 Glutathione S-transferase Mu 2 Not determined. Not determined. Not determined. CX15C phage display library The library was mixed together and added to a well preblocked with nonfat dried milk in PBS. And the library was incubated for 1 hour at RT prior to transfer to GST-M2-2-coated microtiter wells. 1226 HWGMWSY(1) NWGMWSY(1) PHWTWVL(1) HMSKPVQ(1) SSGTHAK(1) YNINIRP(1) NYTQTVP(1) 7 Phage display (common panning) 4 PMID:11748221 HMG box 1 domain of high mobility group protein B1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Peptides were eluted unspecifically by low pH. A computer search of the protein data bases found peptide homologies with proteins already known to interact with HMGB1, like p53, and have allowed us to identify new potential candidates. Among them, transcriptional activators like the heterogeneous nuclear ribonucleoprotein K (hnRNP K), repressors like methyl-CpG binding protein 2 (MeCP2), and co-repressors like the retinoblastoma susceptibility protein (pRb) and Groucho-related gene proteins 1 (Grg1) and 5 (Grg5) can be found. 1227 SSPHNHS(6) HAIYPRH(6) QISFMAN(4) TLTTPIL(2) HWGMWSY(1) ISIPRTM(1) TPAHNDY(1) FHMGQPF(1) APTPVKL(1) HMALNXV(1) MHSLSYR(1) WHWWPXL(1) GETRAPL(1) VQASNSN(1) GNSLRWD(1) SAPQILL(1) DTDPPGL(1) 17 Phage display (competitive panning) 4 PMID:11748221 HMG box 1 domain of high mobility group protein B1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Peptides were eluted by affinity with the same HMG-box used for the biopanning. A computer search of the protein data bases found peptide homologies with proteins already known to interact with HMGB1, like p53, and have allowed us to identify new potential candidates. Among them, transcriptional activators like the heterogeneous nuclear ribonucleoprotein K (hnRNP K), repressors like methyl-CpG binding protein 2 (MeCP2), and co-repressors like the retinoblastoma susceptibility protein (pRb) and Groucho-related gene proteins 1 (Grg1) and 5 (Grg5) can be found. 1228 IQSPHFF(8) LPLTPLP(7) NQDVPLF(2) WPKLASH(2) SPAHAAK(1) ASMSVAI(1) AWLPWAK(1) MPNRTAN(1) NLPAYTS(1) CSSVETH(1) GTPTLXS(1) VMPWVHK(1) YAPRLRS(1) APPTRNQ(1) 14 Phage display (competitive panning) 4 PMID:11748221 HMG box 2 domain of high mobility group protein B1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Peptides were eluted by affinity with the same HMG-box used for the biopanning. A computer search of the protein data bases found peptide homologies with proteins already known to interact with HMGB1, like p53, and have allowed us to identify new potential candidates. Among them, transcriptional activators like the heterogeneous nuclear ribonucleoprotein K (hnRNP K), repressors like methyl-CpG binding protein 2 (MeCP2), and co-repressors like the retinoblastoma susceptibility protein (pRb) and Groucho-related gene proteins 1 (Grg1) and 5 (Grg5) can be found. 1229 HWGMWSY(5) HSWLWWP(5) LAMPQYE(2) KLWVIPQ(1) SHWFWSW(1) HAIYPRH(1) SRPHTSD(1) HYWWWPR(1) SSSSHPT(1) PGAQLTK(1) STTLRYF(1) 20 Phage display (competitive panning) 4 PMID:11748221 HMG box 2 domain of high mobility group protein B1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Peptides were eluted by affinity with the same HMG-box used for the biopanning. A computer search of the protein data bases found peptide homologies with proteins already known to interact with HMGB1, like p53, and have allowed us to identify new potential candidates. Among them, transcriptional activators like the heterogeneous nuclear ribonucleoprotein K (hnRNP K), repressors like methyl-CpG binding protein 2 (MeCP2), and co-repressors like the retinoblastoma susceptibility protein (pRb) and Groucho-related gene proteins 1 (Grg1) and 5 (Grg5) can be found. 1230 ACNMLRWDKTIPCR SCTSPVSQGTQPCK VCPHVAPYQTPLCK RCRKVLICRRICP CCTMPNCTLYVLCE VCEHPENKAPPCCP SCRLSTFSMRMVCN VCPTSEILAGATCR RCGPASREKHALCT VCAKGPPQLPRCP GCAIPQNAIRPPCS SCSPTVYLTSTKCP TCCLCSGKLALACE TCPLPDPDSQYCE RCQALSRLFREKCT FCPGVGRYPVRQCY DCQVRRCSRDSHCE NCQTKVTRITESCC PCSRAETNPTMQCP YCPPPLLNSGEPCS TCKQLKAHPPLACM SCTYSGRADAVVCR 22 Phage display (common panning) 2 PMID:18760481 Anti-WNV E protein monoclonal antibody E16 Envelope protein 1ZTX, Not determined. XC(X)10CX phage display library 1231 CGWASGMADGSSNC CAGWKTGEADGSSC CGWTSGKSDGSASC CTSLVADLDHLSSC CPNIGELSHYDPFC CAAWHTGKAEGNGC CAGRWEHGTAEGDC CTLWVLGLADGTRC 8 Phage display (common panning) PMID:17427229 Trastuzumab Receptor tyrosine-protein kinase erbB-2 1N8Z, Not determined. FMC12C phage display library 1232 CYDGSYRWAPC(3) CYDAQHHWTRC(2) CYNAFLNWVPC(2) CSSPFASC(19) CRLPGSAFTYC(1) CHIPGSIFHLC(4) CYPYGSTFGLC(2) CYAHASNFHMC(2) CRTTGSVFHLC(1) CRWQGSAFAYC(1) CSTSASNYYLC(1) 11 Phage display (common panning) 1 PMID:7685301 Anti-H Fer monoclonal antibody H107 Ferritin heavy chain Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) Comparison of the selected oligopeptides with one another allowed us to derive two consensus sequences characterized by conserved amino acid (aa) residues. Analysis of the distribution of the aa side chains exposed on the surface of H Fer reveals that most of the aa defining both consensus sequences are present either at the end of the big loop or at the end of the A helix. These two regions of the H Fer, though separated in the linear sequence, are very close in the folded molecule. 1233 GRRPNPPIP(4) GRAPNPNLP(2) GRSPGSHMA(5) 3 Phage display (common panning) 2 PMID:7685299 Anti-PTX S1 monoclonal antibody 1B7 Pertussis toxin subunit 1 (PTX S1) Not determined. Not determined. pVIII-9aa phage display library (X9) The selected recombinant phage are capable of mimicking the discontinuous epitope as far as binding to the corresponding mAb, but they are unable to elicit a detectable production of antibodies specific for the original antigen. 1234 CFLRFHPHVLC(1) CRGTSPMDATC(1) CSRTGRCGHGG(1) 3 Phage display (common panning) 2 PMID:7685299 Anti-PTX S1 monoclonal antibody 1B7 Pertussis toxin subunit 1 (PTX S1) Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) The selected recombinant phage are capable of mimicking the discontinuous epitope as far as binding to the corresponding mAb, but they are unable to elicit a detectable production of antibodies specific for the original antigen. 1235 IPKSRKRRIPQR(1) PIQRKRRQMPLS(1) PQLRKRRQNSMM(2) SPQRRKRTRQIR(1) IPSRKRRNRIQQ(1) IKHRKRRLRMIL(1) HMRKRTKIKKIR(1) MMKKTRKRTKRR(2) RKR 8 Phage display (common panning) 3 PMID:20637107 Anti-capsid protein 1 monoclonal antibody C4 Capsid protein 1 Not determined. Not determined. Ph.D.-12 phage display library (X12) The epitope of VP1 is located in the N-terminal and contains the role amino acid sequence R-K-R. Immunization of mice indicated that the phage-displayed peptide induces antibodies against PPV. 1236 APWHLSSQYSRT(6) 1 Phage display (in vivo) 3 PMID:20808875 Mouse cardiac tissue Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A cardiomyoblast cell line, H9C2, was incubated with a M13 phage 12 amino acid peptide display library. Internalized phage was recovered, amplified and then subjected to a total of three rounds of in vivo biopanning where infectious phage was isolated from cardiac tissue following intravenous injection. After the third round, 60% of sequenced plaques carried the peptide sequence APWHLSSQYSRT, termed cardiac targeting peptide (CTP). Authors demonstrate that CTP was able to transduce cardiomyocytes functionally in culture in a concentration and cell-type dependent manner. Mice injected with CTP showed significant transduction of heart tissue with minimal uptake by lung and kidney capillaries, and no uptake in liver, skeletal muscle, spleen or brain. The level of heart transduction by CTP also was greater than with a cationic transduction domain. 1237 KGHSLMP(1) IPTLPSS(1) GTTVIAR(1) QHSEGAL(1) YQDSAKT(1) KVSMNSS(1) LVQPHTR(1) MAHRHPQ(1) QLPGRSG(1) HAIYPRH(1) 10 Phage display (common panning) 4 PMID:20529562 Anti-Human KGF monoclonal antibody Keratinocyte growth factor, KGF Not determined. Not determined. Ph.D.-7 phage display library (X7) Thirty-three out of fifty-eight (56.9%) of the isolated monoclonal phages exhibited high binding activity by ELISA. Ten of fifteen obtained phage model peptides were similar to KGF or epidermal growth factor (EGF). MTT assay data showed that four (No. 1-4) of the ten phage model peptides could promote epidermal cell proliferation. 1238 KHMHWHPPALNT(4) HYQGVHSRYCYH(1) LTPTMFNMHGVL(1) VSRHQSWHPHDL(1) HWPRPDDSFWRP(1) HSTWKLLRLDME(1) GPYFPTHSFLKS(1) 10 Phage display (common panning) 4 PMID:20237096 Nucleocapsid protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The selected phages harboring specific peptides that recognize the N protein of PRRSV were able to efficiently distinguish PRRSV from other viruses in enzyme-linked immunosorbent assays. 1239 APWHLSSQYSRT(3) GYPHPWTLWHLN(1) LDVRPWYVTPLP(1) WAPEKDYMQLMK(1) TPLINMNALTVT(1) P-W-x-[LV] 5 Phage display (subtractive panning) 4 PMID:20711469 Mouse embryonic stem cell line M9 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) To exclude non-specific binding, we used the differentiated ES (dES) cells and primary mouse embryonic fibroblast (PMEF) cells, a feeder layer for the maintenance pluripotency of embryonic stem cells, as a control. Each round of biopanning included three subtraction steps with dES cells, three ubtractions steps with PMEFs and then a selection step with ES cells. Through a phage display screen, authors found phages that displayed an APWHLSSQYSRT peptide showed high affinity and specificity to undifferentiated primate embryonic stem cells in an enzyme-linked immunoabsorbent assay. 1240 VPLYSNTLRYGF SYLTTLRYGNMS TQQSVFSTTLMY IPLPPPSRPFFK QPVDMPYFRTHP TLRYG 5 Phage display (subtractive panning) 4 PMID:20639454 Anti-ARD1A polyclonal antibody N-alpha-acetyltransferase 10, NatA catalytic subunit Not determined. Not determined. Ph.D.-12 phage display library (X12) During each round, the library was preincubated with IgGs purified from normal serum pooled from five healthy volunteers and immobilized on Protein G Sepharose 4 Fast Flow (GE Health care, Uppsala, Sweden) to remove nonspecific binding clones. After the preclearing step, the phage library was selected onto the pool of IgGs purified from patient serum. After four rounds of selection, roughly 1% (10/1000) of phage clones exhibited binding activity to serum from patients with colon cancer. DNA from 10 positive clones that specifically reacted with serum from patients was sequenced. Among 10 phage clones examined, five types of peptides were obtained. The peptide VPLYSNTLRYGF showed significant homology to the residues 121-126 (LYSNTL) of ARD1A by BLAST homology search. Authors revealed that human arrest defective 1 (ARD1A) may serve as an indicator of unfavorable prognosis in colon cancer. 1241 WLSEAGPVVTVRALRGTGSW 1 Phage display (common panning) 6 PMID:15313615 Primary cardiac myocytes (PCM) Not determined. Not determined. Not determined. X20 fAFF1 phage display library The isolated phage, displays the peptide WLSEAGPVVTVRALRGTGSW, and binds these cells 180 times better than a control phage from the library. Furthermore, phage displaying this peptide preferentially bind to cardiomyocytes when compared with a panel of other cell types. A BLAST search revealed that this peptide contains a 12 amino acid segment with sequence identity to a peptide in tenascin-X, an extracellular matrix protein. 1242 GRFLIRVTSSPLGPD(1) TGSGLYLHQMVYLYQ(1) FLIDSPLASIGPTSM(12) FMIDSPLASIGPTSM(2) VIDLSGTRKSSSGTM(1) VRKTTSHPPSYALLH(1) TPPYRAALATPVLLL(1) 7 Phage display (subtractive panning) 3 PMID:20682764 Campylobacter jejuni Not determined. Not determined. Not determined. f88-15mer phage display library (X15) The phage-display library was ubtracted twice against C.jejuni strain ACM 3393 in 3 ml PBSg in a Petri dish, prior to being added to C.jejuni strain C338. Non-binding phages were washed away using PBSg as the wash buffer. Cell surface bound phages were recovered with a low pH elution buffer and amplified by infection in E.coli K91BlueKan cells and used in subsequent rounds of affinity selection. In the second and third rounds of affinity selection, non-binding phage were washed away with PBSg. 1243 FSPFRISELVYTLHP(1) LPFNLAKPELYIFVQ(1) LSAPSPMFLPPVNPH(1) HRPVKTPANAPTTMM(1) CFNDPLDIVPPMLLL(2) 5 Phage display (subtractive panning) 3 PMID:20682764 Campylobacter jejuni Not determined. Not determined. Not determined. f88-15mer phage display library (X15) The phage-display library was ubtracted twice against C.jejuni strain ACM 3393 in 3 ml PBSg in a Petri dish, prior to being added to C.jejuni strain C338. Non-binding phages were washed away using PBSg as the wash buffer. Cell surface bound phages were recovered with a low pH elution buffer and amplified by infection in E.coli K91BlueKan cells and used in subsequent rounds of affinity selection. In the second and third rounds of affinity selection, non-binding phage were washed away with TBST. 1244 FLSDPPAPPTSPGVV(5) 1 Phage display (subtractive panning) 3 PMID:20682764 Campylobacter jejuni Not determined. Not determined. Not determined. f88-15mer phage display library (X15) The phage-display library was ubtracted twice against C.jejuni strain ACM 3393 in 3 ml PBSg in a Petri dish and an extra round of subtraction was carried out against C.jejuni strain C338, prior to being added to C.jejuni strain C338. Non-binding phages were washed away using PBSg as the wash buffer. Cell surface bound phages were recovered with a low pH elution buffer and amplified by infection in E.coli K91BlueKan cells and used in subsequent rounds of affinity selection. In the second and third rounds of affinity selection, non-binding phage were washed away with PBSg. 1245 ITAPHPH NPHAHLQ KPHPHVP TPHLHRD LQPHLHR TPHPHLP TPHPHLR LPHVHSR HQSPWHH HRPPHHW TQPHHTP HAIYPRH AAYTHAR YITHPPH YTFPHWH YWNHNHE SPTHGHD SPSHLHL SPHLHGS SPHLHGA SSPLHVH SSPHNHS ISAHEHL SSLLHVH SKLHMHL KSVQHPH TSLHPHP TGAHVHP EGWHAHT TGAHGHP QFTSLLH QLSHTHI QPQYHTS QSIYVRH QYVRHNH 35 Phage display (subtractive panning, competitive panning) 3 PMID:20863124 Toxin A Transforming protein RhoA Not determined. Not determined. Ph.D.-7 phage display library (X7) Since direct surface immobilization of target proteins can lead to partial denaturation, an affinity capture method was used, immobilizing the recombinant catalytic fragment (rTcdA540) to Ni-NTA resin. A preclearance step was performed prior to +each round of panning to remove plastic and Ni2+ binders from the phage pool. The biopanning protocol was specifically designed to identify those phage that bind rTcdA within the substrate binding pocket by requiring direct competition with RhoA. 1246 MLESHAWPPRAI(3) YISPLPNAATIS(1) TFDRHILDTRGS(1) STVASLGKPTKI(1) ASTIGNLMPGHS(2) FDPHEPTNTRSP(2) LTKEPATGRAML(4) YNKPSFQDHSVI(2) DHIRISTSYKSP(1) TPTRSLDSPHNM(2) DRFTSDLRAPDS(1) 11 Phage display (common panning) 3 PMID:20839946 Phakopsora pachyrhizi urediniospores Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Two peptides, MLESHAWPPRAI and YNKPSFQDHSVI, were identified that inhibit germ tube development when displayed as fusions with the coat protein of M13 phage or as fusions with maize cytokinin oxidase/dehydrogenase (ZmCKX1). 1247 HKQPWYDYWLLR(12) SPPTLYEAWLRF(1) YITPYAHLRGGN(1) SLCLSSAVCWPC(1) SLSSEKLAYWGL(1) 5 Phage display (subtractive panning) 3 PMID:20660187 MLV/HIV 89.6 pseudovirions Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In order to enrich for phage binding to CD4i epitopes, two negative selections were performed, one with MLV/89.6 in the absence of sCD4 and one with sCD4 alone. Each round of positive selection was followed by two consecutive negative selections. During HIV-1 entry, binding of the viral envelope glycoprotein gp120 to the cellular CD4 receptor triggers conformational changes resulting in exposure of new epitopes, the highly conserved CD4-induced (CD4i) epitopes that are essential for subsequent binding to chemokine receptor CCR5 or CXCR4. Due to their functional conservation, CD4i epitopes represent attractive viral targets for HIV-1 entry inhibition. Selected peptides showed partial identity with amino acids in the extracellular domains of CCR5/CXCR4, including motifs rich in tyrosines and aspartates at the N terminus known to be important for gp120 binding. 1248 CPLLSAWPC(2) CKSSSPIWC(1) CPPWYYPRC(1) CQSPDKTHC(1) 4 Phage display (subtractive panning) 3 PMID:20660187 MLV/HIV 89.6 pseudovirions Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In order to enrich for phage binding to CD4i epitopes, two negative selections were performed, one with MLV/89.6 in the absence of sCD4 and one with sCD4 alone. Each round of positive selection was followed by two consecutive negative selections. During HIV-1 entry, binding of the viral envelope glycoprotein gp120 to the cellular CD4 receptor triggers conformational changes resulting in exposure of new epitopes, the highly conserved CD4-induced (CD4i) epitopes that are essential for subsequent binding to chemokine receptor CCR5 or CXCR4. Due to their functional conservation, CD4i epitopes represent attractive viral targets for HIV-1 entry inhibition. Selected peptides showed partial identity with amino acids in the extracellular domains of CCR5/CXCR4, including motifs rich in tyrosines and aspartates at the N terminus known to be important for gp120 binding. 1249 CRPDPGSPC(1) CLPLWPGAC(1) CSSRTMHHC(1) CTRTPAKVC(1) CPNMFSTSC(1) CMSGPPNMC(1) CPNIRAPLC(1) 7 Phage display (subtractive panning) 3 PMID:20802991 B16F10-Nex2 melanoma cell Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Melanoma B16F10-Nex2 cells were used as an affinity matrix for selection of C7C random peptide phage library, and the melanocytic melan-A cell line was used for unspecific phage depletion. After a few rounds of enrichment, 50 phages were randomly selected, amplified, and tested for inhibition of tumor cell proliferation. Seven were active, and the corresponding peptide of each phage was chemically synthesized in the cyclic form and tested in vitro. Three peptides were able to preferentially inhibit the melanoma lineage. A unique peptide, [-CSSRTMHHC-], exhibited in vivo antitumor inhibitory activity against a subcutaneous melanoma challenge, rendering 60% of mice without tumor growth. Further, this peptide also markedly inhibited in vitro and in vivo the tumor cell invasion and cell-to-cell adhesiveness in vitro. 1250 LTGKNFPMFHRN MHRMPSFLPTTL 2 Phage display (common panning) 4 PMID:20890540 Extracellular domain of TβR1 (TβR1-ED) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The output phage from the third round were used as the input for the fourth round without amplification. After four rounds of panning, 24 phage plaques were sequenced and 7 unique clones were identified. Authors evaluated these clones using a phage enzyme-linked immunosorbent assay (ELISA). Two clones exhibited specificity for the target over BSA. 1251 AIWPEPLPLPIG(5) GSNGIWFNLAHR(3) MDSRLGLWPLTW(1) TGLWPSQAQNKA(1) GIWPE 4 Phage display (common panning) 3 PMID:20810747 Anti-E(rns) glycoprotein monoclonal antibody M2172 E(rns) glycoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) 1252 QNWSSAKLPWAP(2) ETWSSVKLPPGI(2) TWQSAKLPWTRP(1) YGWTSGRLPNPP(1) STWPAFRLFTNI(1) YTTTSFRLPNVS(1) YQLRPNAESLRF(1) THSWTSGKIPLR(1) T-x(2)-K-L 8 Phage display (common panning) 3 PMID:20810747 Anti-E(rns) glycoprotein monoclonal antibody M2171 E(rns) glycoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) 1253 TPVALQAKNSAP(1) AVTTQARNIPTV(1) HPLPQARNLPTI(4) TSQQAKNTPTHT(1) LQPQAKNVPTSS(1) EVVYQQSPNTPT(1) V-x(2)-Q-A-R-N-x-P-T 6 Phage display (common panning) 3 PMID:20810747 Anti-E(rns) glycoprotein monoclonal antibody M2165 E(rns) glycoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) 1254 LPYSQSGTLVPP(2) SPLLMSLGGSIV(1) FQYSKAGSFVPE(1) NWSKHGTLLPLV(6) S-x(2)-G-T 4 Phage display (common panning) 3 PMID:20810747 Anti-E(rns) glycoprotein monoclonal antibody M2180 E(rns) glycoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) 1255 APLLVNQSLPHR(1) LTGGTGYSHDFR(2) WAPGSMPTSRLA(2) TTDKHSMTPAAS(1) SHPWNAQRELSV(1) LPFVEWSGISYF(1) QDVHLTQQSRYT(1) TPSLPPTMFRLT(1) L-V-x-G-S-M-P-S 8 Phage display (competitive panning) 3/4 PMID:20452371 Anti-IL-2 monoclonal antibody 1H4 Interleukin-2 (IL-2) Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were competitively eluted from the well with recombinant duck IL-2. Predictive native epitopes L13I14K15G21S23M24P34S41 were determined through aligning the consensus mimotope motifs with duck IL-2 by Clustal W. 1256 KPHTPHNHPSHH(1) KPHHKDIPHSAM(2) KPISHHPHHRAW(1) GPHKHKMTHEHV(4) APHKHHKPPVLM(2) K-P-H-K-H(2)-x-H(2)-S-H-M 5 Phage display (competitive panning) 3/4 PMID:20452371 Anti-IL-2 monoclonal antibody 2B3 Interleukin-2 (IL-2) Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were competitively eluted from the well with recombinant duck IL-2. Predictive native epitopes T33P34T37K38E39C40S41W42Q43T44Y48L49 were determined through aligning the consensus mimotope motifs with duck IL-2 by Clustal W. 1257 THLDKWPKAKPD(3) WPHHKAMPPKIK(5) HSSWWLALAKPT(2) W-x(3)-K-A-K-P 3 Phage display (competitive panning) 3/4 PMID:20452371 Anti-IL-2 monoclonal antibody 5F6 Interleukin-2 (IL-2) Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were competitively eluted from the well with recombinant duck IL-2. Predictive native epitopes W42Q43D67E68K69V70K82P88 were determined through aligning the consensus mimotope motifs with duck IL-2 by Clustal W. 1258 YQDILPNERTQL(2) HDIIERVLPKMP(3) HVLNERYPTSLN(3) IPMEHYPLRKHG(1) VPWELYSLRNRP(1) HVPNERYPLR 5 Phage display (competitive panning) 3/4 PMID:20452371 Anti-IL-2 monoclonal antibody 4G12 Interleukin-2 (IL-2) Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were competitively eluted from the well with recombinant duck IL-2. Predictive native epitopes Y32T33P34N35E58R74F103P104L114R115 were determined through aligning the consensus mimotope motifs with duck IL-2 by Clustal W. 1259 KTVHIGP KTLHIGP CQGKLTC CHGKLTC HQASNFK HLASNYK 6 Phage display (subtractive panning) 3 PMID:20637241 Anti-HIV1 plasma IgG Envelope glycoprotein gp160 Not determined. Not determined. Ph.D.-7 phage display library (X7) Plasma IgG was linked to magnetic microbeads coated with an anti-human IgG. Three rounds of positive selection were performed. The negative selections and amplifications followed the first and second rounds of positive selection. After the third positive selection, individual colonies were picked at random and subjected to analysis by phage capture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hage display (subtractive panning) 3 PMID:20637241 Anti-HIV1 plasma IgG Envelope glycoprotein gp160 Not determined. Not determined. Ph.D.-12 phage display library (X12) Plasma IgG was linked to magnetic microbeads coated with an anti-human IgG. Three rounds of positive selection were performed. The negative selections and amplifications followed the first and second rounds of positive selection. After the third positive selection, individual colonies were picked at random and subjected to analysis by phage capture ELISA. 1261 WLSEAGPVVTVRALRGTGSW 1 Phage display (common panning) 6 PMID:15313615 Adherent primary cardiac myocytes, APCM Not determined. Not determined. Not determined. X20 fAFF1 phage display library The isolated phage, displays the peptide WLSEAGPVVTVRALRGTGSW, and binds these cells 180 times better than a control phage from the library. Furthermore, phage displaying this peptide preferentially bind to cardiomyocytes when compared with a panel of other cell types. A BLAST search revealed that this peptide contains a 12 amino acid segment with sequence identity to a peptide in tenascin-X, an extracellular matrix protein. 1262 GTYNLPNPPPPL(3) KHMHWHPPALNT(3) SAHGTSTGVPWP(3) VPTATLMGASAR(2) WAETWPLAQRPP(2) LSTHTTESRSMV(2) SGHQLLLNKMPN(2) THAAHMGYPSWW(1) LLADTTHHRPWT(2) 9 Phage display (subtractive panning) 4 PMID:20713104 Mouse embryonic stem (ES) cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) To avoid non-specific binding peptides, differentiated mouse ES cells were used as negative target. After four rounds of negative-positive selection, nine sequences in 20 random samples from the chosen clones were selected. Enzyme-linked immunosorbent assay results suggested the peptide (KHMHWHPPALNT) had high affinity and specificity to the mouse ES cells. Immunofluorescence analysis confirmed that peptide (KHMHWHPPALNT) phage selectively bound to the mouse ES cells but not to the differentiated mouse ES cells. 1263 WKVRKSFFKLQG(1) WKLRKWFFKDGG(1) WKAQKRFMKKSG(1) WKVRKAFLFAKN(1) WKMRKSFHKVPG(1) GRKSFHKLDGSF(1) LKTRKLFWKYKD(1) PTTRKWWLKLDG(1) 8 Phage display (common panning) 3 PMID:20833181 Anti-LBP monoclonal antibody M392-2 Lipopolysaccharide-binding protein (LBP) Not determined. Not determined. Ph.D.-12 phage display library (X12) Eight phage clones were found to significantly inhibit the sensitization of LBP on LPS-induced TNF-α release in PBMC. These eight clones had no inhibitory activities on the reinforcement of LPS internalization by LBP, and their displayed peptides indicated the ability to inhibit the inflammatory effects of LBP. 1264 CLHQHNQMC(10) CSRAELSHC(5) CHKSAPTAC(5) CNTSEPRQC(4) CQSDSLSTC(4) CSPTSNSMC(4) 6 Phage display (in vivo) 3 PMID:20863866 Rat lactating mammary gland Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Total 249 peptide sequences were determined from randomly selected individual phage recombinants rescued from lactating mammary tissue after the third round of biopanning. 6 representative peptide sequences were chosen based on their superiority of the appearing frequencies for next study. A peptide ligand, CLHQHNQMC, which specifically homes to the mammary tissue during lactation, was identified through the consecutive in vivo biopannings. The peptide ligand showed specific binding affinity to lactating mammary tissue without any preference to other organs tested in ex vivo and in vivo validation, and microscopy analysis revealed that systemically introduced MG1 could be specifically localized in the lactating mammary gland associated with mammary epithelia and alveolar vasculature. 1265 RNLWPGDLRWVGWHL(1) LGHIWTGRFYAPYRT(1) AREYGTRFSLIGGYR(1) RLGPLHFLNAWGHDH(1) PFYRAGLHSRLGLGG(1) HSAIYYKNFGSSLFR(1) SNLRSWLFPFDRVGN(1) VTGHRWSVRQLGLSH(1) 8 Phage display (common panning) 4 PMID:20609525 Rhipicephalus microplus eggs Not determined. Not determined. Not determined. f88-15mer phage display library (X15) Fourteen individual clones were randomly picked from the eluate of the fourth round of affinity selection and independently propagated for phage production. Eight of the 14 phage clones showed binding on the dot-blots, whereas the remaining six clones no evidence of binding. 1266 CLNSFLRSC CLSTALRSC CSSWYRGAC CTGTSTRAC 4 Phage display (subtractive panning) 4 PMID:20434854 Intact oocytes surrounded by ZP proteins Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) To remove phage clones that bind to the plastic material of test tubes, the primary library was added to an empty test tube prior to incubation with oocytes. Thirty-two random phage clones were analyzed after the fourth round of selection and each clone was sequenced for phage DNA. Sequencing led to identification of three dominant groups of peptide sequences [CLNSFLRSC, CVLSTALRSC (37.5%); CSSWYRGAC (22%); CTGTSTRAC (9%)]. An additional group of peptides represented by only one sequence each (most likely background clones) was isolated as well. 1267 AHPNTAPIHPKF(2) NNQSPILKLSIH(5) WQPVNNAGAILM(1) ATPQNNMMQAQW(1) N(2)-x(2)-P-I-L 4 Phage display (subtractive panning) 4 PMID:20434854 Intact oocytes surrounded by ZP proteins Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) To remove phage clones that bind to the plastic material of test tubes, the primary library was added to an empty test tube prior to incubation with oocytes. NNQSPILKLSIH peptide stimulated production of anti-peptide antibodies. These antibodies bind to the acrosomal region of the canine sperm cell, demonstrating ability to act as sperm antibodies. 1268 DAWKMRLSQMYD VNYNGKEHLLIP IDPLMFKYWSNM SPLNNFYKTQLR VNWNSWHKTNLS AGLPNHNAMLLT NAWLQEPNHRNL TLFTPDKSPAKT SNNADYKQSLLL GALHQEPSSNLF GLFTPDKSPAKT APYDTPWPSPSL CLHCKYTLQQQA GALIYTPEKYTI IDPLMFKYWYNM GPYDTPMFSLNM HAWQSKTPDKTR HTQHSPMVSVEF HVRIPPTMPEAW IDSNHVYKDFLT IPYTHAHADHTL SSKLTFIDFHRN SHDGASSKIRPA SIPTYTPDKVTY SQKYFNDLLDHQ KPVVGMPIVEVW SVPLAKRHLISL SWSSAERLYTMN SYGSSVTQHLAT 29 Phage display (common panning) 3 PMID:20488622 Anti-larval proteins of R. microplus polyclonal antibodies IgG Larval proteins of Rhipicephalus microplus Not determined. Not determined. Ph.D.-12 phage display library (X12) 1269 CTGSWFRPC(1) CLPSWHRFC(2) CLSTGFRAC(1) CHPLRHRPC(1) CVPTWYRAC(1) CLPLTPRSC(1) CSPFNPRVC(1) CTPSWFRWC(1) CSPNWLAHC(1) P-x-W-x-R 9 Phage display (competitive panning) 3 PMID:20594626 Anti-tick subolesin polyclonal antibody (S2) Subolesin Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Sera from non-immunized animals were used as negative control. Phages bound to IgG were sequentially eluted by competition with recombinant tick subolesin ortholog proteins. 1270 CLGPYSFVC(1) CSKLPLALC(1) CNLSRAPFC(1) CLGSYSFIC(1) CSSLDSPMC(1) CYPWGQDHC(1) CPSVLHGWC(1) 7 Phage display (competitive panning) 3 PMID:20594626 Anti-tick subolesin polyclonal antibody (S9) Subolesin Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Sera from non-immunized animals were used as negative control. Phages bound to IgG were sequentially eluted by competition with recombinant tick subolesin ortholog proteins. 1271 CPAAAQALC(1) CPGSFSFIC(1) CGWYSFQHC(1) CGYSFSTSC(1) CWWKPAHLC(2) CPWYSSALC(1) CPAAFAFLC(1) CPSAFHFLC(1) CWYSFSAVC(1) Y-S-F-S-P-x-A-F-S-F-L 9 Phage display (competitive panning) 3 PMID:20594626 Anti-mosquito subolesin polyclonal antibody (S4) Subolesin-like protein Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Sera from non-immunized animals were used as negative control. Phages bound to IgG were sequentially eluted by competition with recombinant mosquito subolesin ortholog proteins. 1272 CPWYLSSLC(1) CISPTDNMC(1) CSLFASAQC(1) CVLPWPWEC(1) CSLNSSAQC(1) CTMPWPFPC(1) CSLPWPFPC(1) CGLGGTAQC(1) CPYGAHWFC(1) S-L-x(2)-S-A-Q, PWP 9 Phage display (competitive panning) 3 PMID:20594626 Anti-mosquito subolesin polyclonal antibody (S5) Subolesin-like protein Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Sera from non-immunized animals were used as negative control. Phages bound to IgG were sequentially eluted by competition with recombinant mosquito subolesin ortholog proteins. 1273 APTPSPGPSYRG(3) SLPTPTPGPWTR(4) YPTPSPGPSARP(2) NTPTGSPGPSTR(1) TPTGQIGPPMRE(1) TPTQSPGPGSRP(1) TPTASFGPTHRS(2) YKPDPTFGPSNR(2) NPDPSPGPTTRH(4) DPPPTFGPHSRS(1) TPPPYWGPHSRD(1) TPSPTMGPPARP(1) QPPHQWGPPSRG(3) LHKPWSPGPSYR(2) QPMPWTAGPTSR(1) IYTPPTWGPPRQ(3) AQPPTPPAGKFR(1) TPTPSPGVLFKT(2) SSVPTPSPGAPF(1) P-T-P-x(2)-G-P-x(2)-R 19 Phage display (common panning) 4 PMID:20630806 Anti-Ap[17-30] polyconal antibody from patient 1 Histone H3-like centromeric protein A Not determined. Not determined. Ph.D.-12 phage display library (X12) 62 phage supernatants of randomly selected bacterial colonies were tested for Ap[17-30] specificity in an ELISA, which identified 36 positive phage clones. 1274 QHSTLPSPGPSL(16) SPTTPSPGPSMK(1) APTPSPGPSYRG(6) AEYRPSPGPSAH(1) NTPSPGPSKAIA(2) FKGQDPSPGPSR(2) TLKPSDGPSRFW(1) DRPSMGPSQFHT(1) STLPSMGPSNFF(1) DANYPSHGPSRY(1) TYLPTKGPSRTV(1) P-[ST]-x-G-P-S 11 Phage display (common panning) 4 PMID:20630806 Anti-Ap[17-30] polyconal antibody from patient 8 Histone H3-like centromeric protein A Not determined. Not determined. Ph.D.-12 phage display library (X12) 62 phage supernatants of randomly selected bacterial colonies were tested for Ap[17-30] specificity in an ELISA, which identified 36 positive phage clones. 1275 GHPDPWLIEAPS HHDPWDRLERFT VHDPWSNNSTWN SHDPWDLLPLPY SHDPWDEGPQRA AHSERWERFPQG H(2)-D-P-W-D-x-LP 6 Phage display (competitive panning) 3/4 PMID:20619902 Anti-IL-2 monoclonal antibody 2E6 Interleukin-15 Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were competitively eluted with recombinant goose IL-2. 1276 QLETFSIKNMPR TSIPHIMYLDPF FKWPTTEWPSRL NFMESLPRLGMH SLTVPFLPLYVP ASRQDMQNLAPP E-S-L-S-R-x(2)-M-x(2)-L-x-P 6 Phage display (competitive panning) 3/4 PMID:20619902 Anti-IL-2 monoclonal antibody 3A1 Interleukin-15 Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were competitively eluted with recombinant goose IL-2. 1277 WAPGSMPTSRLA HLPTSSLFDTTH SHPWNAQRELSV ATWSHHLSSAGL DRYNSEWLPYSP SSPHHMVSYTWL S-H(2)-L-P-T-S-x-L 6 Phage display (competitive panning) 3/4 PMID:20619902 Anti-IL-2 monoclonal antibody 3B3 Interleukin-15 Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were competitively eluted with recombinant goose IL-2. 1278 VPVIWDVPYQSL MHNPWDAFMEAS GHPDPWLIEAPS SHDPWDLLPLPY MHPDPWDAPESR YHSDPWSGFLKD KPVYWDHELSKS H-P-D-P-W-D-A-P-L-S(2) 7 Phage display (competitive panning) 3/4 PMID:20619902 Anti-IL-2 monoclonal antibody 5A9 Interleukin-15 Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were competitively eluted with recombinant goose IL-2. 1279 TLKPTQASLVHG HVGTGDGHEHWQ QLETFSIKNMPR HSNWRVPSPWQL GIHELNVARLRL H-E-P-W-Q-L-x-L 6 Phage display (competitive panning) 3/4 PMID:20619902 Anti-IL-2 monoclonal antibody 5C3 Interleukin-15 Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were competitively eluted with recombinant goose IL-2. 1280 CFRHMTEQC(6)[0.545] CIPLPFYNC(5)[0.22] CSHLYLHNC(2)[0.464] CPLTGTSKC(1)[1.04] CTLKVRGEC(1)[0.97] CGFWHTSKC(1)[0.411] CAWPSKDNC(1)[0.887] CIPLLFHNC(1)[0.893] CWGAMVHPC(1)[0.452] CHASLKHRC(1)[0.785] CYTPWMPRC(1)[2.01] CTYLAPKGC(1)[0.851] 12 Phage display (common panning) 4 PMID:19027991 Beta-amyloid protein 42 (Beta-APP42) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA To determine the apparent dissociation constant (Kd, nM), the amyloid-beta 42 coated wells were incubated with twofold serial dilutions of each phage clone, starting with 1.0e12 virions(1.66e(-8) M) in the first well and ending with 8.0e6 virions (1.33e(-13) M) in the last well. Kd of the 12 selected clones displaying the highest affinities for amyloid-beta 42 was shown. Data shown were reproduced from the graph. Two phage clones expressing the peptides C-IPLPFYN-C and C-FRHMTEQ-C were strongly selected because they are present in several copies. The first peptide is characterized by five hydrophobic amino acids out of seven suggesting that it interacts with Aβ42 by hydrophobic bonds. It is interesting to note that Aβ42 itself aggregates by hydrophobic mechanisms inside AP structures. On the other hand, peptide C-FRHMTEQ-C is hydrophilic, so the interaction mechanism should be differ ent and could involve the hydrophilic N-terminus domain of Aβ42. 1281 TSYTTSIFGPRA(1) ILANDLTAPGPR(1) NYMSALAITTSL(1) KPANLTSTPWVP(1) LLADTTHHRPWT(1) ISMPVRPLLQDF(1) SIVSTQTSLPLN(1) SMTSHNQWHLLA(1) ANPSPSTHHLTP(1) TTTLLTATPHPH(1) YPSFTHSATPSL(1) FHQNSLRVHSSP(1) QDVHLTQQSRYT(1) NLNHERSQNLKM(1) YESIRIGVAPSQ(1) ERVMLPFPPAPW(2) IPWTQHMAMSPM(1) TNTSWMTAMTPF(1) THTTNAEGYSPV(1) TMGFTAPRFPHY(1) SVSVGMKPSPRP(4) SINGQWMRAIGK(1) GIQLANPPRLYG(1) QDMLKPYVDPLH(1) SHHIPSYQWPLH(1) WAETWPLAQRPP(1) SNQTSDRPPLLT(1) SSLEPWHRTTSR(2) EWLAYDGIRAYS(1) ETLPITLIARLT(1) 30 Phage display (subtractive panning) 5 PMID:20501662 Glycine receptor subunit alpha-1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Human Embryonic Kidney (HEK) Cell were transfected with GlyR α1 subunit cDNA. Phage were applied to blank (control) HEK 293 cells. Then, phage that did not bind in this negative selection procedure were removed from the plate with a pipette, applied to the plate of GlyR-expressing cells. 1282 DHARYPWLRPPA LPAFFVTNQTQD YWNASPSASGVI WHTEILKSYPHE NSPRLVHTNTHN LLADTTHHRPWT SHVDDLGLRPLT 7 Phage display (common panning) 3 PMID:20544195 Anti-EGFR monoclonal antibody 12H23 Epidermal growth factor receptor Not determined. Not determined. Ph.D.-12 phage display library (X12) Genomic DNA from 20 phage clones was sequenced, and seven different insert sequences were observed. Two (WHTEILKSYPHE and LPAFFVTNQTQD) mimotope candidates were specifically recognized by the selecting antibody 12H23 but not by the isotype-matched control antibody. 1283 ASGALSPSRLDT(13) 1 Phage display (subtractive panning) 4 PMID:20570932 Human 143B osteosarcoma cell lines Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Before each selection round, a negative selection with the 293T human embryonic kidney cell line was done to subtract phages that bound to nontumor cells. After the final selection round, the ssDNAs of 20 clones were sequenced and analyzed. One sequence was found to be enriched in 65% of all sequenced clones. Peptide ASGALSPSRLDT shares a significant homology with heparinase II/III family protein, which binds and reacts with heparan sulfate proteoglycans. 1284 SVSVGMKPSPRP(8) NHNTSTWAAYST(1) TLPSPHSLLTVH(1) 3 Phage display (common panning) 3 PMID:20329714 GaAs semi-insulating substrate Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The specific adhesion of these peptides is used for biofunctionalization of GaAs/AlGaAs photonic waveguides capable of enhancing the second harmonic generation response. 1285 YLCKFGC(1) SPDELHK(2) REPLVYW(1) AEPIIYW(2) KQPIVFW(1) 5 Phage display (common panning) 4 PMID:20844088 Anti-SEB monoclonal antibody ab53981 Enterotoxin type B (SEB) Not determined. Not determined. Ph.D.-7 phage display library (X7) The N-terminal 15-mer peptide of SEB was determined to be an epitope of ab53981. 1286 MGGTLIASDQYQ(17) QSLPASMSYQTA(1) SLSASMDFMMYA(1) NALRASNSFMDE(1) L-x(2)-S 4 Phage display (common panning) 4 PMID:20844088 Anti-SEB monoclonal antibody ab53981 Enterotoxin type B (SEB) Not determined. Not determined. Ph.D.-12 phage display library (X12) The N-terminal 15-mer peptide of SEB was determined to be an epitope of ab53981. 1287 AHPWWLG(4) SHPWYLG(2) P-x(2)-L-G 2 Phage display (common panning) 4-6 PMID:20652782 Anti-S100A4 monoclonal antibody 22.3 Protein S100-A4 Not determined. Not determined. Ph.D.-7 phage display library (X7) The consensus sequence of MAb 22.3 mimotope was corresponded to amino acids 43-47 of S100A4 amino acid sequence. 1288 AWPDKQP(1) SWPDKMP(1) ILSDKMA(1) PDKQP 3 Phage display (common panning) 4-6 PMID:20652782 Anti-S100A4 monoclonal antibody 22.1 Protein S100-A4 Not determined. Not determined. Ph.D.-7 phage display library (X7) The consensus sequence of MAb 20.1 was corresponded to amino acid 94-98 of the C-terminal part of S100A4. 1289 HGINTTKPFKSV WPSSYLSPIPYS AVTTLTLVPAGT ALTGSTKSAFGG 4 Phage display (common panning) 6 PMID:20405826 Na+ Montmorillonite (MMT) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1290 HGINTTKPFKSV TSVRTHFPLYPV GFEHKDQRFTRE 3 Phage display (common panning) 6 PMID:20405826 Primary Ammonium C18 Modified Montmorillonite (MMT) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1291 HGINTTKPFKSV GTFPLAIKARDP LHLPSDRLRLWG MHRSDLMSAAVR 4 Phage display (common panning) 6 PMID:20405826 Quaternary Ammonium C18 Modified Montmorillonite (MMT) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1292 LSVTYCFVY NSIWGLFVH PVNMTFVAL RFSTYVSSW VHFSYVSVV RYSCARSTS WYARFNLAA WVRFVGVTG GRWVALLDV SVARFLVVD RVRPARYLG YVQVARYLS RFLGMRRGF VRWLAWKMV LRYIMALAV GTRFALETS WDRWAMSTG RYWVVIETV SKGARFWLR RWGFRFFAG FSRYFSLRL YSRWEVRFM AWVRFWEDD RYLEIADFK LRVSFLDIA FVSLWLEVA AWGVYLDLG WGVYLEAVS YVTGYLDVV GRVFFWDLE MVGFWELVR PPRGWKLEW VSLYSSDGL VVGYLMEVL VSLFMGIDF VYTRLEGCL VVAFLMGQD QGFLSFLRE WAWLMATME WIFYSLSTE WVLPVQWSM VGFLFNLLW YVFRMRLVS VVFLAAGSY WLSLLNVRV MIGYAVRPH FWAFALARA EPYMLGVTR GTCYMSVVS VRMMCWYPV EARVLACLF 51 Phage display (common panning) 5 PMID:20338912 Chymase Not determined. Not determined. Not determined. X9 T7 phage display library Nonapeptide-coding inserts were then sequenced from 75 individually sampled phages. Fifty-one of the obtained sequences were aligned. Seventeen background sequences that had also been observed in analyses of other enzymes were excluded. Eleven of these contained the insert WCQVQSVCA and six the insert TLMVPRTGS. Four sequences contained stop codons. The remaining three sequences contained repetitive sequences and did not contain aromatic aa. 1293 RLQLKL(2) RTRYED(3) RIPLEM(3) QFDEPR(3) TSAVRT(2) QCTGRF(1) ITDMAA(1) WRPCES(1) 4 Phage display (in vivo) PMID:20460372 Breast cancer tumors Not determined. Not determined. Not determined. pComb M13 phage display library 1294 RLQLKL(11) GLWQGP(7) QCTGRF(5) LPGMMG(5) DVGTTE(5) TDLGAM(5) GMMYRS(4) DSNAES(4) ITDMAA(3) RWRTNF(4) WRPCES(2) WRNTIA(3) IDKQLE(3) FMEIET(3) HEVVAG(2) GGHTRQ(2) INGKVT(2) 17 Phage display (subtractive panning) 6 PMID:20460372 Mixed liver/kidney tissue extracts Not determined. Not determined. Not determined. pComb M13 phage display library Phage library was pre-exposed to extracts of liver and kidney, tissues that typically have high nonspecific uptake of drugs and imaging agent, to allow cleavage by proteases contained within these extracts. Isolated phage were reamplified and subjected to six additional rounds of both negative (no liver/kidney cleavage) and positive (tumor cleavage) selection, with representative phage being isolated and sequenced after each round. 1295 QQSWPIS(4) 1 Phage display (common panning) 3 PMID:20648291 Pd wire Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Peptide QQSWPIS appeared four times among ten colonies. The peptide can bind to the Pd NC surface and act as a stabilizer to mediate Pd crystal nucleation and growth. 1296 MFQRMAVDA(1) YRQMSAPTL(1) WFGQLQAAQ(1) YRHAQAYGV(1) MPRQYTQMI(1) FHKVYRGLL(1) WQMAMSRHL(1) SIMMMGAVR(1) WYTLLKSRL(1) YKPMLASVG(1) YAAMRGGQI(1) YKPLWGQMT(1) NYQMMGTMR(1) FERARSRLL(1) YLRMLQILK(1) NWRALRGAL(1) YRQMWVNRA(1) FMAMRAVRL(1) FTMLMNQVM(1) VYRAQGPQM(1) MMSRMLGQT(1) SSGRYARLG(1) TFYRMGTQM(1) WIGQMYAAK(1) YRMQSNHVS(1) DRSMEWRMM(1) FSAIRNRIL(1) YRAMQTTLS(1) NYLGMKPRV(1) RGFRHMMAA(1) YQQMGARLM(1) SWRHIQTRV(1) MYGLMLAQR(1) YQALRAYWQ(1) RFTQMMAVA(1) WKGLQGALS(1) FMRLGGGHL(1) IWHMQNARV(1) YKLLRATQM(1) LWQSFMQMA(1) FHQLRGGSL(1) YRGMQRRTL(1) YGKLRGGTL(1) QVGKYLGLG(1) TYRGMAGFR(1) RVFQNLRAS(1) WRQMYRAQL(1) RSYSLAPGR(1) TWNSLRGRL(1) QRYNAMRAA(1) YIMQMNRVV(1) LTLMQAKRL(1) RFTQMGALA(1) YYGKMPALV(1) PWQAMRGWL(1) KKIMSYHAL(1) YRGMSAFRA(1) MFHRMSGLQ(1) GQRSYSSMQ(1) YMMQKSVRA(1) 60 Phage display (common panning) 4 PMID:1402647 HLA class II histocompatibility antigen, DRB1-1 beta chain HLA class II histocompatibility antigen, DM Not determined. Not determined. M13lib X9 phage display library DR1 molecules were isolated from the human EBV-B cell line, HOM-2. M13 phages were incubated with biotinylated DR1 in binding buffer (50 mM Tris-C1, pH 7.5, 150 mM NaC1, 1 mM EDTA, 1 mM PMSF, and 0.2% NP-40). After at least 24 h incubation at room temperature, BSA-blocked streptavidin on 4% beaded agarose was added and incubated for 10 min. The M13 phage/DR1 complexes were purified by washing the solid phase several times with binding buffer. Competition experiments with both isolated phages and corresponding synthetic peptides showed that the binding was specific. 1297 EPLQLKM QQQLSTH 2 Phage display (common panning) 5 PMID:20942387 TIMREX SLP30 Primary Synthetic Potato graphite Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Modification of graphene with bifunctional peptides reveals both the ability to impart selective recognition of gold nanoparticles and the development of an ultrasensitive graphene-based TNT sensor. 1298 TMGFTAPRFPHY YHRMPQALSAME 2 Phage display (common panning) 5 PMID:20942387 TIMREX SLP30 Primary Synthetic Potato graphite Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Modification of graphene with bifunctional peptides reveals both the ability to impart selective recognition of gold nanoparticles and the development of an ultrasensitive graphene-based TNT sensor. 1299 GAMHLPWHMGTL 1 Phage display (common panning) 5 PMID:20942387 AFM standard highly ordered pyrolytic graphite (HOPG) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Modification of graphene with bifunctional peptides reveals both the ability to impart selective recognition of gold nanoparticles and the development of an ultrasensitive graphene-based TNT sensor. 1300 GAMHLPWHMGTL 1 Phage display (common panning) 5 PMID:20942387 N006 nano graphene platelets Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Modification of graphene with bifunctional peptides reveals both the ability to impart selective recognition of gold nanoparticles and the development of an ultrasensitive graphene-based TNT sensor. 1301 CRGWRGEKIGNC CNMQALSMPVTC 2 Phage display (competitive panning) 5 PMID:20129923 Bifunctional heparan sulfate N-deacetylase/N-sulfotransferase 1 Heparin Not determined. Not determined. CX10C T7 phage display library The library was biopanned against a chimera of mNdst1 consisting of the full-length protein minus the cytoplasmic tail and transmembrane domain fused to protein A. mNdst1 was immobilized on IgG beads, and heparin was used to displace bound phage. Both peptides inhibited mNdst1 activity in vitro, however, by distinct mechanisms. The peptide CRG WRGEKIGNC presents a chemokine-like repeat motif (BXX, where B represents a basic amino acid and X is a noncharged amino acid) and binds to heparan sulfate, thus blocking the binding of substrate to the enzyme. The peptide NMQALSMPVT inhibits mNdst1 activity by direct interaction with the enzyme near the active site. 1302 SNFYMPL(48) 1 Phage display (subtractive panning) 4 PMID:20637198 Human OE33 (esophageal adenocarcinoma) cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Peptide selection was performed using phage display by removing nonspecific binders using Q-hTERT (intestinal metaplasia) cells and achieving specific binding against OE33 (esophageal adenocarcinoma) cells. After completion of biopanning, plaques from a total of 60 phage that demonstrated preferential binding to the OE33 cells were selected. Authors found that 48 of these 60 clones (80%) contained the identical DNA sequence and corresponded to the peptide with amino acid sequence SNFYMPL. The other 12 clones expressed different unique amino acid sequences. The peptide sequence SNFYMPL binds specifically to dysplasia in Barrett's esophagus and can be fluorescence labeled to target premalignant mucosa on imaging. 1303 HKTTLSPASH(1) HKTASTPATH(1) PRFSTTPASH(1) AKGNTAPATD(1) NNSSAAPATQ(1) RFPTAAPATA(1) TKSYIAAPAS(5) TXSYIAAPAS(1) GKYFTAAPAS(1) LKTLMTAPAT(1) FPDPPTAPAT(1) LARPNVAPAT(1) FSPLNWPSAT(1) AATPRAYAPAS(1) ARPAPATYG(1) HNMAPATLH(1) HPWKQEAAPAS(1) LRPAPASVI(1) HCAAPASRQ(1) APATHLPAQ(1) VTASPATLFG(1) SSPPASIHSL(1) I-A(2)-P-A-[ST] 22 Phage display (common panning) 3 PMID:20699234 Lupus-derived monoclonal antibody BT164 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After three panning rounds a consensus was obtained with sequence IAAPAS/T corresponding to amino acids 27-KSAPAT-32 from histone H3. 1304 HTDWRLGTWHHS(3) NWITMNPAMPTL(6) LPFNTLADPRIN(5) LLADTTHHRPWT(5) NLLIPENVPLRH(1) 5 Phage display (common panning) 5 PMID:20949958 Hyperbranched poly(phenylene vinylene) (hypPPV) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1305 ELWSIDTSAHRK(6) YYPASSTIQSRP(2) HPTLHMTYYKKQ(1) HIHRGEHGPSAR(5) TPLTPNGLTRSG(2) IVKNVPLTPLRE(1) AVPHRVGGLHSL(1) NYLHNHPYGTVG(1) EHPHVPITPSNL(1) 9 Phage display (common panning) 5 PMID:20949958 Linear poly(phenylene vinylene) (linPPV) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1306 AFTHEAPKRRDS(4) SFPGPASARAGA(13) QIEESFVRGJTT(1) SLYKLRNVAAGR(2) CRWSTPTTSQCP(1) IHWSWTTVERPH(3) WPANILSHGPKH(1) 7 Phage display (common panning) 5 PMID:21131596 Apoptosis regulator Bcl-2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) At the final round, bound phages were eluted by addition of excess amount of Bcl-2 protein. Authors showed that the P6 peptide specifically recognized binding to the Bcl-2 protein in the cell system (in vivo). 1307 EHWSHDMFSPGD(9) EVSLHDMNLATH(2) KWLGHDMIMSGT(1) FNTKHDMQGDTS(1) EHNDFPMYTWRP(1) TTTHFLATKFYK(1) E-x(3)-H-D-M 6 Phage display (common panning) 3 PMID:21118490 Anti-urease B monoclonal antibody U001 Urease subunit beta Not determined. Not determined. Ph.D.-12 phage display library (X12) EXXXHDM matched the urease B proteins at 347-353 aa. 1308 CEKLKHSMC(10) CTKTWQSMC(2) CMSLLGHKC(1) CKHGLLSMC(1) CMSQDGHTC(1) E-x(3)-H-S-M 5 Phage display (common panning) 3 PMID:21118490 Anti-urease B monoclonal antibody U001 Urease subunit beta Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) EXXXHSM matched the urease B proteins at 347-353 aa. 1309 STLMTTTYHSVS(1) QGAHYEYSRTEL(1) IPGDAGKGLHMT(1) YDTTSSPPRLTR(1) GMIKAAHERPLR(1) VLSNSLPTAIST(1) GMATEATVHELA(1) LIRGLLIGFGRN(1) YPLLPESPTDAN(2) NWSAEKAKLYDS(1) QNSHRVALENNT(1) 11 Phage display (common panning) 3 PMID:21234986 Indium nitride (InN) semiconductor (SC) material Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Prior to the phage display, the InN surfaces were preliminarily prepared by appropriate chemical etching; thus, native oxides have been removed assuring the specific phage to reach the SC. As the YPLLPESPTDAN appears two times over 12, this peptide can be expected to be a good binder. 1310 SATTHYRLQAAN(11) TEHPSNTSPMRL(2) SGNTHYRLQAAN(1) TPHRLDWSPHLV(1) 4 Phage display (common panning) 4 PMID:21151669 Fibrosarcoma cell line NG4TL4-tk Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A 12-mer amino acid peptide SATTHYRLQAAN, denominated TK4, was isolated from a phage-display library with fibrosarcoma tumor-binding activity. In vivo biodistribution analysis of TK4-displaying phage showed a significant increased phage titer in implanted tumor up to 10-fold in comparison with normal tissues after systemic administration in mouse. 1311 CAYHRLRRC(55) CGFYWLRSC(5) CSFFYLRSC(16) R-L-R(2), F-[FY]-x-L-R-S 3 Phage display (common panning) 4 PMID:21063027 Human leukemia cells MOLT-4 Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Phage selection was performed with an excess of the competing Arg-Gly-Asp (RGD) synthetic integrin-binding peptide motif to minimize and eliminate the recovery of RGD-containing ligands. By screening of human leukemia cells with a combinatorial phage display peptide library, Authors isolated a peptide motif, sequence Phe-Phe/Tyr-Any-Leu-Arg-Ser (FF/YXLRS), which bound to different leukemia cell lines and to patient-derived bone marrow samples. 1312 CAVTDGKQC(5) CQRPPR(4) CRPAR(4) CRPPR(2) CKPPR(1) CKAPR(1) CGRQGAR(2) CTLTGNSKC(2) CASLVLVAC(2) CKPPTPEEC(1) CRPAVRISC(1) CSYTKDKTC(1) CSHDA(1) CQSVRG(1) 14 Phage display (subtractive panning) 2-3 PMID:20869411 Macrophages isolated from primary human lung squamous carcinoma Not determined. Not determined. Not determined. Byungheon Lee (Department of Biochemistry and Cell Biology, School Medicine, Kyungpook National University, Republic of Korea) For subtraction, the eluted phage solution was incubated with cells from normal tissue and then the supernatant containing unbound phages was recovered. Using phage display, Authors identified the CQRPPR peptide, named ApoPep-1(Apoptosis-targeting Peptide-1), that was able to home to apoptotic and necrotic cells in tumor tissue. ApoPep-1 also bound to apoptotic and necrotic cells in culture, while only little binding to live cells was observed. Its binding to apoptotic cells was not dependent on calcium ion and not competed by annexin V. 1313 TIPKWISIIQALR(94) 1 Phage display (common panning) 3 PMID:21093050 Peptide SLGLSFYQPPEGDNALC Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The selected phage population was screened further by affinity chromatography using PMCA from rabbit duodenal mucosa which expressed mostly PMCA1. Following competition, 184 plaques were picked and their plasmid DNA was sequenced. The dominant phage clone had 94 copies and encoded for a variable peptide sequence TIPKWISIIQALR. 1314 SFHQLPARSPLP GSTQAWMSPPLA LLADTTHHRPWT 3 Phage display (subtractive panning) 3 PMID:20952686 In vitro M cell coculture system Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative panning was performed to remove the phage particles displaying peptides capable of binding to the monocultured Caco-2 cells. On the basis of the deduced amino acid sequences of these peptides, authors selected three peptides because these sequences were found at least four times and were not found in the sequences from phage particles that bound to monocultured Caco-2 cells. One of the selected peptide ligands, SFHQLPARSPLP, promoted the binding of ligand-fused Ag to mouse Peyer's patch M cells and human M-like cells that had been defined by binding with the M cell-specific and anti-GP2 Abs. 1315 QTYTPTKQSPPMPRP YLRITSTPHKYLTQR IKAPNPPSVSTLPPR RKRAPPPALRTAPRS PRDSMLFMPCKFRPM PVLIFKKIVLTAPVA AKPLASPRETGASQR RKSLPIALQLHHSYN QSPSCHMSSLQCTQL KLVCVSANFQSGVPH SRLTRNDMVTGCMC HVTSTKAWYDPSQPL RAKASATVCHLSNPY PMRPKGPDTLLYTLP VILNWRPSCLVTCHE KEMVVSCPPGALAYS MNRLPDFTRLPPYSP ANGVPIPPHPARAAR KGMRMRLAIGPDSA RPKLLHCEPSPAVVQ QSGMPQCYTALRPRV TPGLDAAVFWSLGSE LVIAPMCKPPPYTPE QHTLIIDATMPSASP RVMKHRTPSSRYQKP TNSSRPLSKSRQYRC KDPALLTRSPPSARM TSMIYRPHNVQTLLH APSGFARRTQTFWYV 29 Phage display (subtractive panning, in vivo) 4 PMID:20958260 PC-3 human prostate carcinoma cell Not determined. Not determined. Not determined. f88-15mer phage display library (X15) The library was precleared of phage that bound to the vasculature and other non-tumor antigens in normal non-tumor bearing CF1 mice, and then utilized in an in vivo selection scheme in PC-3 human prostate tumor-bearing mice. 1316 INTECAGLGLVCKPHT NKSKCRCRQNACKQLI CTSSCKPHSQSCKEKT NKKQCKTVLKMCHRRV DRPHCLKTWNICTSYY MKRECKNRCALCKSER IVPGCSKTERGCSYQS DMASCTQHADNWLKHE KPSPCSSFKSHCVRRD AKYYCEELVNHCTSAQ GDLRCRITKQKCEQQC ETIMCIRYRCDCPLPH GPAHCKRTISQCQTNE DEWHCKFNGAVCTSMR TPILCENIRSGCELKR QRVTCDMAENCCPKTS TNTECNTLSKLCQSSI HMQVCATVTHQCWIYG PPRLCQGMRGTCSGNQ CACICPCNPAFCTVAV KMPECHEQQEYCDGDR QKEHCILHTANCGRIT TNTNCGTDLEPCVSTM LKGDCTQRYVYCMKSK HDKQCLTAKDRCGTIK GDSSCQEVQTQCRYSK ASCECNPHPRHCGETR 27 Phage display (subtractive panning, in vivo) 4 PMID:20958260 PC-3 human prostate carcinoma cell Not determined. Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) The library was precleared of phage that bound to the vasculature and other non-tumor antigens in normal non-tumor bearing CF1 mice, and then utilized in an in vivo selection scheme in PC-3 human prostate tumor-bearing mice. 1317 RAPMGGR RRPVGRA RLPPKAG RRPVVGR RAPARRY SRPGLRR VGPRTLR RGPRGRV 8 Phage display (common panning) 5 PMID:21071869 Soluble 37/48kDa oligomer formation of Aβ(1-42) Not determined. Not determined. Not determined. T7 phage display library (X2PX4) Although the reason for the enrichment of the arginine residues is not fully understood, highly cationic sequences containing at least three arginine residues are probably important in binding to the soluble Aβ(1-42) structure, because arginine contains a basic guanidine group. These peptides were found to suppress specifically 37/48kDa oligomer formation and to keep the monomeric form of Aβ(1-42) even after 24h of incubation, as disclosed by SDS-PAGE and size-exclusion chromatography. 1318 CGSYGFNAC(3) CSLERLGFC(7) CYRLTGLWC(2) CLDSASRGC(2) CGLRLESTC(2) CWLFSVSAC(2) 6 Phage display (common panning) 2-3 PMID:21070862 Virulent isolate of Paracoccidioides brasiliensis (Pb18) Not determined. Not determined. Not determined. CX7C fUSE5 phage display library Variability of CGSYGFNAC phage binding to different Pb isolates were examples of variability of expression by the fungus of its binding molecule, strongly suggesting CGSYGFNAC as a biomarker of virulence. 1319 CTSGTHPRC 1 Phage display (common panning) 4 PMID:21182881 Primary rat lung alveolar epithelial primary cell cultures Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The first three-rounds identified apical cell-surface binders, while the 4th round aimed to identify phage internalised by the ATI-like cells. Peptide CTSGTHPRC showed significant pulmonary epithelial translocation across highly restrictive polarised cell monolayers. Cell biological data supported a differential alveolar epithelial cell interaction of the CTSGTHPRC peptide-phage clone and the corresponding free synthetic CTSGTHPRC peptide. 1320 GWWYKGRARPVSAVA(15) RAVWRHSVATPSHSV(7) LWRPVLFHSAVRALG(1) WRGVYFGDRWLGSQP(1) GWYSSRHYVRSLNGL(1) QQLVYNWWAVSSARR(1) LSWPLHAGRGFRWVS(1) 7 Phage display (common panning) 4 PMID:19558186 Ganglioside GM3 Not determined. Not determined. Not determined. fUSE5-based X15 phage display library The peptides were found to have affinity for GM3, and alanine scanning showed seven amino acid residues that contribute to carbohydrate recognition. The binding of peptides to the cell surface was significantly inhibited in the presence of sialic acid or by the digestion of cell surface sialyl residues by neuraminidase. 1321 EPWLDSRYSPLS GHGLLQYTDVMF TSLYTDRPSTPL LPIPSSLGGPFP SYEMPFSTRPWF LPGWPLAERVGQ TGHQSPGAYAAH NFMESLPRLGMH HSTWKLLRLDME HATGTHGLSLSH SFLYSYTGPRPL TMGFTAPRFPHY SLKMPHWPHLLP KAWIVQPPFHYS QLPKHNYWPGAF YTTSNTLQVIAR NTPGIRPQATYS ALHPLTNRHYAT SNTSIIRNAFPQ GVSQNTNSLHLR GMVSTSRMHAGW 21 Phage display (common panning) 4 PMID:15278181 Germania Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The powder was exposed to the phage library, after which the powder was washed repeatedly to remove any free phage particles. The bound phage were then eluted by incubation glycine-HCl solution. The 3 most dominant peptides identified from the clones were chosen for further evaluation. The extent of germania precipitation (i.e. the germania precipitating activity) was quantified by adapting the β-silicomolybdate colorimetric assay. SLKMPHWPHLLP and TGHQSPGAYAAH peptides exhibited relatively high germania-precipitating activities, whereas the TSLYTDRPSTPL peptide exhibited much lower activity. 1322 NLLMAAS KLWVIPK HHHRHSF HPWLTRH 4 Phage display (common panning) PMID:14978510 Extracellular domain of Tie2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In fact, the target is Tie2-Fc. At the end of the selection, 48 clones were isolated and sequenced, showing that 11 different peptides were represented. Each selected clone was assayed by ELISA for binding to Tie2-Fc. Four clones gave the highest signal. One peptide, NLLMAAS, completely abolished the binding to Tie2 of both angiopoietin 2 and angiopoietin 1 (Ang1). We further show that NLLMAAS specifically suppresses both Ang1-induced ERK activity and migration in human umbilical endothelial cells. Moreover, in vivo, this peptide inhibits angiogenesis in the chick chorioallantoic membrane assay. 1323 GALTPLR(1) SYTPLRP(1) GFTPHRS(1) AIETLIR(1) NYYTPHR(1) HTQTRHR(1) NDLTPLR(1) SPFTPHR(1) SYTPHRT(1) T-x(2)-R 9 Phage display (common panning) 3 PMID:18400009 Anti-M2e monoclonal antibody 8C6 Matrix protein 2 Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage clones exhibited a consensus motif (TXXR), which was found on the epitope EVETPIRN on influenza A virus M2 protein. Site-directed mutation analysis indicated that Thr and Arg on the epitope EVETPIRN played a key role in the recognition by 8C6. Furthermore, sequence alignment and analysis revealed that Thr and Arg on the epitope were highly conserved. 1324 WGRFWGRWLA(5) VCDWWGWGIC(3) YWMGWKWEGE(6) WWDFLQGSER(1) FAMWYPLGWR(1) W-x(2)-W 5 Phage display (competitive panning) 5 PMID:9492279 Doxorubicin P-glycoprotein Not determined. Not determined. fUSE5-based X10 phage display library To ensure a high level of the drug absorption on polysterene, doxorubicin was conjugated to BSA, and the conjugate was immobilized on polystyrene ELISA plates. Displacement with verapamil, a competitive inhibitor of doxorubicin binding with P-glycoprotein were used to elute the phages that bound to the doxorubicin-BSA2-coated plates. The displacement assay showed that the phages expressing these peptides bound MDR type drugs (vinblastine, doxorubicin, verapamil, and genistein) with the same selectivity as P-glycoprotein and did not interact with non-MDR type drugs, such as arabinosylcytosine (Ara-C) and melphalan. The structure modeling suggested that all the selected sequences contained a hydrophobic envelope in which MDR drugs could be docked with substantial energy minimization. 1325 TWWWTWAGKH(1) LWSPWYGGSW(9) W-x(2)-W 2 Phage display (common panning) 4 PMID:9492279 Doxorubicin P-glycoprotein Not determined. Not determined. fUSE5-based X10 phage display library To ensure a high level of the drug absorption on polysterene, doxorubicin was conjugated to BSA, and the conjugate was immobilized on polystyrene ELISA plates. Treatment with low-pH buffer which is usually used for phage panning were used to elute the phages that bound to the doxorubicin-BSA2-coated plates. The displacement assay showed that the phages expressing these peptides bound MDR type drugs (vinblastine, doxorubicin, verapamil, and genistein) with the same selectivity as P-glycoprotein and did not interact with non-MDR type drugs, such as arabinosylcytosine (Ara-C) and melphalan. The structure modeling suggested that all the selected sequences contained a hydrophobic envelope in which MDR drugs could be docked with substantial energy minimization. 1326 HEIGSLAGLAMR(1) AHFNGSLRSLTQ(1) IINTGSLLSLAH(1) DTGSLVWLSQRS(1) HNHGSISALMHL(1) DHDRLIRRTAQI(1) FHTLDNNIRNVN(1) FHDTSLLSHHLA(1) G-S-L-x(2)-L 8 Phage display (common panning) 4 PMID:20961714 Anti-FMDV Asia1 antigen monoclonal antibody 3E11 Polyprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) Through a 12-mer random peptide phage display, synthetic peptide analysis and constructing a series of FMDV Asia1/YS/CHA/05 mutants using reverse genetic system, we finely mapped the neutralizing epitope as the 12-amino acid peptide 141SXRGXLXXLXRR152. 1327 ELKQSAQVQAKI HLWSFALPSAQV INSARIMPPNWR TSPQARLEAIEP YSLRQDYNPEPV YSLRQDVLVLEE YSLRNDWPTLPP SLTNSMFPGYAY HMSIWPERWPLS HSLRPEWRMPGP FQIAPTDPASNA SPKIISSGLPTY TSSGLAPTPIMQ FGTGLASHSPAW HNLVPRHLGTVL SYQWHHLGTWLT STYGSHLGWGHF GIHLNFKVTHMH WHSTMSWTNSTW AGHDRLAVTPSS LTRSLDSHSMNT 21 Phage display (common panning) 3 PMID:21071249 Methylmalonic aciduria and homocystinuria type C protein Methylmalonic aciduria and homocystinuria type D protein, mitochondrial Not determined. Not determined. Ph.D.-12 phage display library (X12) The MMACHC-affinity-selected peptides were scanned against MMADHC to predict binding sites of MMACHC on this protein. Five distinct regions of multiply-aligning clusters were located along the primary sequence of MMADHC. Two regions were positioned in a domain with homology to ATPase component of a bacterial ATP-binding cassette (ABC) transporter: Region I, residues142-150; and Region II, residues 157-169. The other three regions were located in the C-terminal domain of MMADHC: Region III, residues 220-236; Region IV, residues 246-256; and Region V, residues 280-290. 1328 LLDSARH GPASGPM TPSSNRF AHTPNSR 4 Phage display (common panning) 3 PMID:21071249 Methylmalonic aciduria and homocystinuria type C protein Methylmalonic aciduria and homocystinuria type D protein, mitochondrial Not determined. Not determined. Ph.D.-7 phage display library (X7) The MMACHC-affinity-selected peptides were scanned against MMADHC to predict binding sites of MMACHC on this protein. Five distinct regions of multiply-aligning clusters were located along the primary sequence of MMADHC. Two regions were positioned in a domain with homology to ATPase component of a bacterial ATP-binding cassette (ABC) transporter: Region I, residues142-150; and Region II, residues 157-169. The other three regions were located in the C-terminal domain of MMADHC: Region III, residues 220-236; Region IV, residues 246-256; and Region V, residues 280-290. 1329 NMNKHPLAYTEP(1) FHWSWYTPSRPS(5) WHFEWWRATPSG(1) VLPPKPMRQPVA(1) HLQSMKPRTHVL(1) 5 Phage display (common panning) 4 PMID:21214859 Glutathione S-transferase, GST Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1330 CHLYTAGSCNMS(2) CHLSATGACRMI(5) IGSDMKGMPKPR(1) KSLSRHDHIHHH(1) 4 Phage display (common panning) 4 PMID:21214859 Cytoplasmic domain of CORYNE Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In fact, the target is recombinant glutathione S-transferase(GST)-CRN-His6. Two positive clones that specifically bind to the intracellular protein kinase domain of CRN, CHLYTAGSCNMS and CHLSATGACRMI, have been identified. Alignment of these peptides and the kinase-associated protein phosphatase (KAPP) shows high similarity, indicating that KAPP might interact with the cytoplasmic kinase domain of CRN and negatively regulate the CLV signal. 1331 SYTFHWHQSWSS(3) QSWSWHWTSHVT(3) WTWRWAHVTNTR(1) QDVHLTQQSRYT(1) HKAHEYDPWISP(1) SYSQHYGIPNPW(1) SSWQMSWSWMGS(1) QSWS 7 Phage display (common panning) 5 PMID:21259034 Dry degummed silk fibroin fibers Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The selected silk-binding peptides contained a consensus sequence QSWS which is important for silk-binding as confirmed by binding assays using phage and synthetic peptides. 1332 EHYLDKWASHDM(1) LPQLNTLDKWAT(2) ASSLDKWALYTS(2) AAMTNHVLDKWA(1) YKLDKWAPSLSK(1) YPTLDKWAMWMH(1) KHLALDKWSILA(1) LPHLDKWSSITP(1) ITPALDKWSTRF(1) QVPLDKWSVAWP(1) MDLDKWNMKSLT(1) SLDKWVLAPLAY(1) NGLDISLDKWGV(1) HPLTLLDKWTLL(1) QSHYDKWASWGF(1) ALSEDKWASTAS(1) YGFPYDKWASRP(1) NMDKWAAVFQSK(2) YLPDKWATPQMI(1) ATNYDKWALPYT(1) EHAYDKWAQRXX(1) TPDKWALPHPTL(1) QFIPDKWARSPN(1) AIQLVYDKWAMP(1) HLPYTSDKWALM(1) YMDKWANIVALR(1) SHPGPDKWQTLP(2) TPDKWYGLAPYR(2) TPDLWSYLSNLQ(1) TNLFDKWSYLAS(1) YPNIDKWVALYH(1) ANDPDKWNLTPL(1) SPIHDKWSDLSR(1) TPFLDKWVALKP(1) YLPTDKWSHLRT(1) SWPFDKWSQSLS(1) SLHETHMLDRWA(1) TTLQTLDRWSQL(2) LESSLDRWTTLA(2) QTMDRWASLRWS(1) LVPDRWAFLQMS(2) MPDPDRWALWPL(1) ALVSDRWSWMHQ(1) ETLAKDRWVTLG(1) DPWAFRWPSGPM(1) ELDKWAS 45 Phage display (common panning) 3 PMID:21237206 Anti-HIV-1 gp41 MPER monoclonal antibody 2F5 HIV-1 gp41 membrane proximal exterior region (MPER) Not determined. Not determined. Ph.D.-12 phage display library (X12) 1333 CKTLDKWAC(4) CMALDKWAC(2) CWHLDKWAC(2) CWTLDKWAC(1) CLVLDKWAC(1) CLRLDKWAC(1) CVSLDKWAC(1) CLGNDKWAC(1) CDKWAPPTC(5) CDKWAGLLC(4) CDKWAPTSC(3) CDKWALAYC(2) CDKWAPRSC(1) CDKWAVRWC(1) CDKWAPPSC(1) CDKWAEQYC(1) CDKWASQPC(1) CDKWAFMTC(1) CDKWAPPGC(1) CDKWAPSAC(1) CDKWAPSWC(1) CDKWAQTYC(1) CDKWATRFC(1) CDKWAGRAC(1) CDKWAGPRC(1) CDKWAPRQC(1) CDKWAIPWC(1) LDKWA 27 Phage display (common panning) 3 PMID:21237206 Anti-HIV-1 gp41 MPER monoclonal antibody 2F5 HIV-1 gp41 membrane proximal exterior region (MPER) Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 1334 SNFFDRTWPKLT YNFFDRTWPKLT SHRQHETDRNWP NYPEDFFQRTWP F(2)-D-R-T-W-P 4 Phage display (common panning) 3 PMID:21264311 Anti-DENV2 monoclonal antibody 2A10G6 Dengue virus 2, DENV2 Not determined. Not determined. Ph.D.-12 phage display library (X12) After three rounds of panning, 94 positive phage clones were identified and subjected to DNA sequence analysis. The results showed that 86 of the positive clones displayed 12 amino acid residues in common, i.e., SNFFDRTWPKLT. Phage-display biopanning and structure modeling mapped 2A10G6 to a new epitope within the highly conserved flavivirus fusion loop peptide, the 98DRXW101 motif. 1335 APRDPLS(1) DPFFYTP(1) GLDHHPP(1) GYDKHPQ(1) HFTNHPQ(2) HLANHPQ(1) HLENHPM(2) HLIAHPQ(1) HLYAHPQ(2) HRISWPS(1) HYEGHPQ(1) IAPNHPQ(2) IPEWHPQ(8) IPYWHPQ(3) ISSTHPQ(1) MQSHQDS(1) NLISHPQ(1) NLLNHPQ(11) NLVNHPQ(4) NLVSHPQ(1) NPTKHQM(1) PLLAHPQ(1) SLIAHPQ(4) SLLAHPH(1) SLLAHPQ(7) SLLSHPQ(2) SLVSHPM(1) SPTYQRL(1) TLIAHPQ(1) TLISHPQ(5) TLLAHPQ(8) TLLNHPQ(2) TLQPGGA(1) TPSPLAG(1) VTPTMHP(1) YLVNHPQ(1) YMEHSRV(1) HPQ 37 Phage display (common panning) 4 PMID:21278676 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. Ph.D.-7 phage display library (X7) One hundred phage isolates were sequenced after the fourth round of selection against streptavidin-coated beads. 1336 HHHPPLA(5) HHPPFPP(2) HPPSWGD(2) HPPHFPN(1) HPTGLFR(2) HNHLGVH(2) KPFHNST(5) KPGYSSA(2) KPPQVPL(2) KPHAPHR(1) KPPHHPR(3) KPPHPVY(1) KPVKVPR(2) 13 Phage display (common panning) 4 PMID:21278676 Unmodified helix 31 of bacterial 16S ribosomal RNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Duplicate samples of unmodified h31 on magnetic beads were employed for selection. For each sample of unmodified h31, 100 isolates were sequenced. Two hundred phage isolates were sequenced after the fourth round of selection against unmodified h31 on streptavidin-coated beads. Two hundred phage isolates were sequenced after the fourth round of selection against unmodified h31 on streptavidin-coated beads. Only peptides with repeated sequences or common motifs are shown. 1337 FVRPFPL(6) FVRPFAL(4) FVRPYAP(2) FPRTIAP(1) DIRTQRE(4) DIRTQTR(2) DIRATQA(2) ATPLWLK(5) ATPTQRE(3) ATPLYLR(2) TLWDLIP(5) TLWSFMP(3) TLWVPSR(2) TLTTLTN(2) TLTFFHR(2) TYLPWPA(5) TYLPWPP(2) TYLRARL(3) TYPFAPW(2) TLW, VRP 19 Phage display (common panning) 3 PMID:21278676 Modified helix 31 of bacterial 16S ribosomal RNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Duplicate samples of h31 on magnetic beads were employed for selection. For each sample of h31, 100 isolates were sequenced. Phage isolates were sequenced after the third round of selection against wild-type modified h31 on streptavidin-coated beads. Two hundred phage isolates were sequenced after the third round of selection against wild-type modified h31 on streptavidin-coated beads. Only peptide sequences with repeated sequences or common motifs are shown. Several of the peptides exhibited moderate affinity (in the high nM to low μM range) to modified helix 31 in biophysical assays, including surface plasmon resonance (SPR), and were also shown to bind 30S ribosomal subunits. These peptides also inhibited protein synthesis in cell-free translation assays. 1338 CVRPFAL(5) CVRAPTL(2) TVRPFTL(2) TLWDLIP(2) TLWPLSP(2) TLW, VRP 5 Phage display (subtractive panning) 3 PMID:21278676 Modified helix 31 of bacterial 16S ribosomal RNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) A separate counter selection was also carried out in the third round of selection against wild-type h31 in which free unmodified h31 was added to the phage and modified h31-streptavidin mixture. One hundred phage isolates were sequenced after the third round of selection. Only peptide sequences with repeated sequences or common motifs are shown. Several of the peptides exhibited moderate affinity (in the high nM to low μM range) to modified helix 31 in biophysical assays, including surface plasmon resonance (SPR), and were also shown to bind 30S ribosomal subunits. These peptides also inhibited protein synthesis in cell-free translation assays. 1339 SDSLMKKPGGPG(1) MYHHRKKPGGPG(1) MGAMHKKPGKTG(1) VHKYDKKRGKRG(1) PTGKMKKAGGLL(1) EPPRYKKHGGAS(1) AHHTLLELWTQA(1) ASYSNLTHWLSS(1) SPRLTTYKASPK(1) DMHHKPGRTLHH(1) K(2)-P-G(2)-P-G 10 Phage display (common panning) 3 PMID:21375771 Anti-WNV C protein monoclonal antibody 6D3 Capsid protein C Not determined. Not determined. Ph.D.-12 phage display library (X12) The mAb 6D3 recognized the phages displaying a consensus motif consisting of the amino acid sequence KKPGGPG, which is identical to an amino acid sequence present in WNV C protein. Further fine mapping was conducted using truncated peptides expressed as MBP-fusion proteins. 1340 AYYPQNHKSNAE 1 Phage display (common panning) 5 PMID:21329494 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1341 WHWRNPDFWYLK(0.225) WHWTWLSEYPPP(0.215) LETSKLPPPAFL(0.125) WHWSQWLSGSPP(0.085) WHRTPQFWAFPW(0.07) SVSVGMKPSPRP(0.05) WHKTPWFWPTNL(0.05) WHWSWQPQRHSP(0.04) WHWQYTPWWRGS(0.03) 9 Phage display (common panning) 5 PMID:21329494 Gliadin Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Panning was done both with gliadin dissolved in urea and in NaHCO 3. Since electrostatic binding of the phage to the target protein weakens with increasing ionic strength, which in turn influences the specificity of the interaction, different buffers with different ionic strengths were tested prior the actual panning experiments. Although identical sequences were frequently picked up in independent experiments, altogether more than 160 unique sequences encoding peptides with potential gliadin binding activities were identified. In order to confirm that the selected phage clones interacted with gliadin proteins, nine different phage populations that had repeatedly been picked up in different panning experiments were chosen. Together, these nine sequences represent 89% of all identified gliadin-binding sequences. 1342 HAPVQPN(1) CTGPTSLSC(2) CKPMQFVHC(1) CSSYEYHAC(1) CSTQAHPWC(1) CGTSRLFSC(1) CASHNPKLC(1) CPAKQKAHC(2) CSASGTPSC(1) CTRFYRPSC(1) CQNPRQIYC(4) CNPQMQRSC(5) CNYPTLKSC(7) NPYHPTIPQSVH(4) HQFISPEPFLIS(1) SPNFSWLPLGTT(2) SVSVGMKPSPRP(2) TPLTSPSLVRPQ(1) 18 Phage display (competitive panning) 4 PMID:21291244 Single-crystal hydroxyapatite (HAP) Not determined. Not determined. Not determined. Ph.D.-12, Ph.D.-7 and Ph.D.-C7C phage display library pool In last round, the HAP binding phages were simultaneously exposed to the HAP target substrate in solution. 1343 CNERQMELC(1) CNKPLSTLC(1) CHTLLSTTC(1) CLKPFSGAC(1) CLGPGKAFC(4) CSTSAKHWC(1) TMGFTAPRFPHY(7) 7 Phage display (competitive panning) 4 PMID:21291244 Polycrystalline hydroxyapatite (HAP) Not determined. Not determined. Not determined. Ph.D.-12, Ph.D.-7 and Ph.D.-C7C phage display library pool In last round, the HAP binding phages were simultaneously exposed to the HAP target substrate in solution. 1344 HWKHLHNTKTFL 1 Phage display (common panning) 4 PMID:21295090 Amino acid (160)REVPYAYIREGHEKQ(174) of MMP-14 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Peptide HWKHLHNTKTFL (denoted as MT1-AF7p) showed high MT1-MMP binding affinity. Computer modeling verified that MT1-AF7p binds to the MT-loop region of MT1-MMP and interacts with MT1-MMP through hydrogen bonding and hydrophobic interactions. 1345 TYTDWLNFWAWP(1) 1 Phage display (common panning) 5 PMID:21297616 Myelin basic protein, MBP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the selection against MBP, phage expressing a library of peptides were selected through multiple cycles for binding to biotinylated MBP, using avidin agarose to isolate selected phage. Specifically, the phage library was mixed with biotinylated MBP and allowed to bind for 1 h. Avidin agarose was added and incubated for an additional hour. Nonbinding phage were removed by washing the agarose three times with PBS solution and the supernatant was plated for titer and amplification for subsequent cycles. 1346 NTQTLAKAPEHT(5) KSLSRHDHIHHH(2) DFTKTSPLGIH(2) 3 Phage display (subtractive panning) PMID:21297616 Excised murine peripheral nerves Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Specifically, for positive selection using nerve tissue, nerve tissue was dissected/washed and mixed with a phage library. For negative selection using nonnerve tissues, nonnerve tissues (muscle and fat) were dissected from normal mice and incubated with the phage library obtained from the positive selection. 1347 AHHNSWKAKHHS(1) 1 Phage display (in vivo) 8 PMID:21297616 Nerve tissue (sciatic, brachial plexus, cranial nerves) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1348 CSTSMLKAC(79) CKPGTSSYC(20) CPDRSVNNC(18) 3 Phage display (in vivo) 3 PMID:21316369 Ischemic region of left ventricular (LV) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The CSTSMLKAC sequence was capable of mediating selective homing of phage to ischemic heart tissue. 1349 CWEPMNHLC(14) CPFASYLHC(13) CENVWYPRC(9) 3 Phage display (in vivo) 3 PMID:21316369 Non-ischemic region of left ventricular (LV) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The first two rounds of bio-panning were performed against ischemic left ventricular (LV) tissue. 1350 CQSTMSTLC(13) CPTSFLTDC(9) CYDLRSHQC(7) 3 Phage display (in vivo) 3 PMID:21316369 Right ventricular (RV) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The first two rounds of bio-panning were performed against ischemic left ventricular (LV) tissue. 1351 CTESAPYFC(1) CPDANNGNC(1) CNMAQTNMC(1) CPNANLGTC(1) 4 Phage display (common panning) 3 PMID:21329515 Intact matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The blocking strategies with bovine serum albumin (BSA) were deployed, and eluates treated with acid were used. 1352 CINSFYAQC(1) CKSAISSSC(1) CVPQYSSQC(1) CQPKAVNHC(1) CPVSPSGAC(1) CSNASRPFC(1) CNPALSTHC(1) CGKAGLPLC(1) CPTHPPFQC(1) CESSAIRYC(1) CNNGTSRLC(1) CPSQTHPTC(1) CTNQQRHTC(1) 13 Phage display (common panning) 4 PMID:21329515 Intact matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The blocking strategies with bovine serum albumin (BSA) were deployed, and eluates treated with acid were used. 1353 CPPTPLSLC(1) CINASKPLC(1) CNRMVQPMC(1) CNLALTQAC(1) CQEPRSNAC(1) CPSHHLESC(1) CNPLHRQHC(2) CSKTFPVRC(1) CFKHSSHQC(3) 9 Phage display (common panning) 3 PMID:21329515 Solubilized matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The blocking strategies with bovine serum albumin (BSA) were deployed, and eluates treated with acid were used. 1354 CFKHSSHQC(2) CTYPFHASC(1) 2 Phage display (common panning) 3 PMID:21329515 Solubilized matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The blocking strategies with bovine serum albumin (BSA) were deployed, and eluates treated with acid/salt were used. 1355 CLSTSSKSC(9) CQTSANTQC(1) CGVPAGSTC(1) CLATKLHNC(1) CDGVSTKHC(1) CIKNPTKYC(1) CMPSPSLKC(1) 7 Phage display (common panning) 3 PMID:21329515 Solubilized matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The blocking strategies with bovine serum albumin (BSA) were deployed, and eluates treated with glaycosaminoglycan (GAG) were used. 1356 HWHDWMWSWRRD(2) SMWPWYYSQWAR(1) TLGDRYSTKHPI(1) SFSTMNTAPGGS(1) WYMPWWSAGQAA(2) QKKIRKRPHVKR(1) GAFHKHHHARLI(1) WNRSPLPDYGAA(1) SLWQRWFPVLDH(1) DLALRNPTPSDP(1) SHALPLTWSTAA(1) WHYNSWYRWPVM(1) 12 Phage display (common panning) 3 PMID:21329515 Solubilized matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The blocking strategies with bovine serum albumin (BSA) were deployed, and eluates treated with acid were used. 1357 AYYPQNHKSNAE(2) APQYQHNQATHT(1) SITWTHHPGALQ(1) AGLHPRSLESLP(1) HPGNRSLDPLNH(1) LLADTTHHRPWT(1) ATGKPTRLESHV(1) NPSNLYRQPAMT(1) SKAHDISQRQPP(1) VNRIPGENLSSP(1) SNQPAPALFHQL(1) YSPASKSPVPSL(1) SVSVGMKPSPRP(2) 13 Phage display (common panning) 3 PMID:21329515 Solubilized matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The blocking strategies with bovine serum albumin (BSA) were deployed, and eluates treated with glaycosaminoglycan (GAG) were used. 1358 CVPSSARIC(1) CRPHDSKAC(1) CHPEPRSQC(1) CVEKRPRQC(1) CFMDYRNLC(1) CSHSVQPFC(1) CQTHNPRQC(1) CDGAPAPLC(1) CKTDLQKQC(1) CGPFPQPHC(1) CSFHGPGPC(1) CSTNQTPTC(1) CSTSPONSC(1) CKLIHNNSC(1) 14 Phage display (common panning) 3 PMID:21329515 Cryostat-sectioned matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The blocking strategies with bovine serum albumin (BSA) were deployed, and eluates treated with acid were used. 1359 CHMSPRHQS(1) CLPNKQWSC(1) CKQPLNNTC(1) CTVTPRHLC(1) CDNTSKTQC(1) CASTTAACC(1) CLHMDKKRC(1) CMKTPMRSC(1) CYKHVGQRC(1) CHLSPFKSC(1) CTTSKYRDC(1) CTATGLSNC(1) CPSSMPSRC(1) CPATSHTHC(1) 14 Phage display (common panning) 3 PMID:21329515 Cryostat-sectioned matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The blocking strategies with bovine serum albumin (BSA) were deployed, and eluates treated with acid/salt were used. 1360 CVQSSTQHC(1) CSGHHSLRC(1) CPTSQQKVC(1) CNSTHPRAC(1) CNRLESHLC(1) CTNPHRSQC(1) CTKTPWPGC(1) CNRLQGEHC(1) CPTPTGRYC(1) CVPTAMSNC(1) CSLARPNEC(1) CVRTPFSMC(1) CNNTTPPSC(1) CTSQQKANC(1) 14 Phage display (common panning) 3 PMID:21329515 Cryostat-sectioned matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The blocking strategies with phosphate-buffered saline with Tween (PBS-T) were deployed, and eluates treated with acid were used. 1361 CHVTAQRAC(1) CATPEWPPC(1) CPNLMNTRC(1) CTKSSPPRC(1) CTQTTVASC(1) CDQSKTIAC(1) CSRGSMGIC(1) CSPIRGSMC(1) CSHTGHHQC(1) CHEPTTMAC(1) CSRADLTTC(1) 11 Phage display (common panning) 3 PMID:21329515 Cryostat-sectioned matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The blocking strategies with phosphate-buffered saline with Tween (PBS-T) were deployed, and eluates treated with acid/salt were used. 1362 CKGPVSRHC(1) CTTSSEHVC(1) CSTTMKTSC(1) CDNKRSPAC(1) CKLNYPNAC(1) CQFSKSQSC(1) CTLDTRRDC(1) CPFSSSPSC(1) CPSMSHHQC(1) CSASTQSFC(1) CPLKGLATC(1) CTGKPLKTC(1) CIHMTGYHC(1) CEMTETKHC(1) CKENWPLIC(1) CSNSPTTMC(1) 16 Phage display (common panning) 4 PMID:21329515 Cryostat-sectioned matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The blocking strategies with bovine serum albumin (BSA) were deployed, and eluates treated with acid were used. 1363 CGKHDDTYC(1) CPTKDLRYC(1) CTSSGNRYC(1) CTIKTNLQC(1) CHSTAKSAC(1) CPASKGDFC(1) CSHRVPHDC(1) CHATPYPKC(1) CDSSRHTHC(1) CSRLSQEYC(1) CTGKQYPQC(1) CGMNAFRAC(2) CTTKYSTTC(1) CSSDKALVC(1) CSPRSHLSC(1) CPPSPMPYC(1) 16 Phage display (common panning) 4 PMID:21329515 Cryostat-sectioned matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The blocking strategies with phosphate-buffered saline with Tween (PBS-T) were deployed, and eluates treated with acid were used. 1364 HETFPSPRANSV(6) SQIDYATGPRQA(1) WDTEKASPLSPL(1) DHTGKSPGLFHN(1) 4 Phage display (common panning) 4 PMID:21329515 Solubilized matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The blocking strategies with non-fat milk (NFM) were deployed, and eluates treated with acid were used. 1365 AHKHKHPGHITA(1) 1 Phage display (common panning) 4 PMID:21329515 Solubilized matrices (Integra ®) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The blocking strategies with phosphate-buffered saline with Tween (PBS-T) were deployed, and eluates treated with acid were used. 1366 STSVLYN THHHLPN AFHGPVH LPLTPLP 4 Phage display (common panning) 4 PMID:21362346 Anti-human TGF-β1 monoclonal antibody Transforming growth factor beta-1, TGF-beta-1 Not determined. Not determined. Ph.D.-7 phage display library (X7) The data of MTT showed that TGF-β1 and one phage model peptide (LPLTPLP) could promote keloid fibroblasts proliferation, however, other three phage model peptides could inhibit keloid fibroblasts proliferation. 1367 ELMAVPVPLPPA(5) SEYTSQLIFTAT(3) SEFSYIVIDTSL(4) ELTAILVSPAPL(8) ELNAQHIMEPKY(10) ELIPMLIMQSTS(1) EDYSTIMKTLAH(1) STPKSPHSVASH(1) AVQHNPTHPFYP(1) AHSTGLSPSTLR(1) TMSSVAPRNLSS(1) 11 Phage display (common panning) 3 PMID:21364990 Candida albicans, C. albicans Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The peptides can distinguish C. albicans from other closely related species. 1368 QPHHYSL(1) NPHSYPH(1) QPHHYPL(1) SPHHYPH(1) TPHHYMH(1) QPHHYFK(1) WPHHFPH(1) VPHGYFL(1) TPHGYAH(1) VPHSYPH(2) QPHHYPF(1) APHHYPM(1) LAINIKS(1) SPHSYPR(1) P-H-x-Y 14 Phage display (subtractive panning) 4 PMID:21399663 Patient CA502 ascitic IgG Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The library was pre-absorbed with the total immunoglobulin G (IgG) mix of 10 healthy donors to remove the phages recognized by the IgG. 1369 NGTTSSNNQLINENNIQN EHMYNTPHTYHTTMKNNK QPIHPNNM NKLAAALE KNYKN TNTHN KHTNN 7 Phage display (in vivo) 3 PMID:21408169 Mouse colonic dysplasia Not determined. Not determined. Not determined. X18 T7 phage display library 1370 NGTTSSNNQLINENNIQN EHMYNTPHTYHTTMKNNK NKLAAALE KNYKN TNTHN KHTNN 6 Phage display (in vivo) 4 PMID:21408169 Mouse colonic dysplasia Not determined. Not determined. Not determined. X18 T7 phage display library 1371 WHKEQFW(9) NTWHNSR(12) THSPGLL(1) SINPDNR(1) HVQLLIF(1) SITTVAA(1) ALLADSR(1) EIALGAR(1) AGPTRIS(2) TVKTRPA(2) ATSAIHG(1) QTKMTNP(2) 12 Phage display (subtractive panning) 3 PMID:21418812 Purified lgG of tuberculosis serum Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Purified IgG of health people serum was used as the molecule of counter selection during the second and third selection. Twelve phage clones with different sequences were amplified and detected with indirect ELISA and single phage HVQLLIF showed higher affinity with IgG of tuberculosis and Was identified as the positive clone. 1372 CPKGLDWCC(7) 1 Phage display (common panning) 3 PMID:21423621 Protein E7 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The binding specificity and affinity of the selected peptide to HPV16E7 were tested by competitive enzyme-linked immunosorbent assay (ELISA). The antagonist peptide showed obvious anti-tumor efficacy both in cell lines and animal tumor models. 1373 VPQQKFR(2) HSQISSY(1) QPTHQLT(1) QHQLPSD(1) SLLSTPQ(3) LRPTLSC(1) LPMRPIV(1) H-x-Q, L-x-S 7 Phage display (common panning) 3 PMID:12970876 Anti-gastric cancer antigen monoclonal antibody MGb1 Gastric cancer antigen Not determined. Not determined. Ph.D.-7 phage display library (X7) These 10 phage clones could also partly inhibit the binding of MGb1-Ab to gastric cancer cell KATO-III. 1374 CHASIYDFGSC(18) CVYALIMPPLC(1) 2 Phage display (common panning) 5 PMID:10760806 Anti-CCR5 monoclonal antibody 3A9 C-C chemokine receptor type 5 Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) All clones were tested by capture ELISA for reactivity with mAb 3A9 and 23 of 26 clones showed strong reactivity. After sequencing, 19 nonameric peptide inserts could be identified. For phage CHASIYDFGSC, the motif SIYD aligned to residues at the N terminus and FG to residues on the first extracellular loop. 1375 CPHWLRDLRVC(15) CLPPSYCFGSC(1) CPPVFGTFTSC(1) CYGPFSRASYC(1) CMPPSMTSVSC(1) 5 Phage display (common panning) 3 PMID:10760806 Anti-CCR5 monoclonal antibody 5C7 C-C chemokine receptor type 5 Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) After three rounds of biopanning, 64 phage clones were isolated and tested by capture ELISA for reactivity with mAb 5C7. Nineteen clones showed very strong reactivity and sequencing revealed five different phagotypes. For CPHWLRDLRVC, residues at the N terminus, first extracellular loop, and possibly the third extracellular loop could be aligned and so would contribute to the mimotope. 1376 AQNPSDNNTHTH ARYDLSIPSSES HGNPLPMTPFPG RLELAIPLQGSG 4 Phage display (common panning) 3 PMID:10864319 Semiconductor crystalline surface GaAs(100) Not determined. Not determined. Not determined. X12 M13 phage display library 1377 ASSSRSHFGQTD WAHAPQLASSST TPPRPIQYNHTS GTLANQQIFLSS 4 Phage display (common panning) 4 PMID:10864319 Semiconductor crystalline surface GaAs(100) Not determined. Not determined. Not determined. X12 M13 phage display library 1378 VTSPDSTTGAMA AASPTQSMSQAP SSLQLPENSFPH 3 Phage display (common panning) 5 PMID:10864319 Semiconductor crystalline surface GaAs(100) Not determined. Not determined. Not determined. X12 M13 phage display library 1379 MYWYPY(2) IPWYPY(2) LYWYPY(6) RIFYPY(2) PIFYPY(1) LPFYPY(1) FYWYPY(1) YVYYPY(5) FYYYPY(1) PLFVRY(1) TYSATV(1) WFSFMS(2) ENGRKS(1) SSSGFW(1) VPGVSF(1) ARRYSR(1) HSSYFF(1) RRHHHH(2) LHHHH(1) HWLVHH(1) HRHKHQ(1) WPDMVR(1) FNAAVL(1) YPY 23 Phage display (common panning) 4 PMID:1376919 Concanavalin-A, Con A Methyl α-D-mannopyranoside Not determined. Not determined. f3-6mer phage display library (X6) Biotinylated ConA (bio-Con A) at 2μM in Tris-buffered saline (TBS) was incubated on streptavidin-coated polystyrene Petri dishes, for a total of four rounds of biopanning. 1380 MYWYPY(8) IPWYPY(7) LYWYPY(1) IAWYPY(3) IFWYPY(1) VWWYPY(2) TAFQLS(1) HRVGGT(1) CACRLK(1) YPY 9 Phage display (common panning) 4 PMID:1376919 Concanavalin-A, Con A Methyl α-D-mannopyranoside Not determined. Not determined. f3-6mer phage display library (X6) In the first round, biotinylated ConA (bio-Con A) at 2μM in Tris-buffered saline (TBS) was incubated on streptavidin-coated polystyrene Petri dishes. The samples of amplified phage were subjected to remaining rounds of biopanning at 0.2μM bio-Con A. 1381 MYWYPY(11) YTWYPY(1) PYWYPY(1) VGRAFS(1) VSWYIS(1) RAAGIV(1) YPY 6 Phage display (common panning) 4 PMID:1376919 Concanavalin-A, Con A Methyl α-D-mannopyranoside Not determined. Not determined. f3-6mer phage display library (X6) In the first round, biotinylated ConA (bio-Con A) at 2μM in Tris-buffered saline (TBS) was incubated on streptavidin-coated polystyrene Petri dishes. The samples of amplified phage were subjected to remaining rounds of biopanning at 0.02μM bio-Con A. 1382 SLERSI KQVKLN VWGVFP SLRWGS DLRSAF IARRAI RVIPMY FIPLFS RFIWRW REMVFY FLMVRT 11 Phage display (common panning) PMID:7588601 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. f3-6mer phage display library (X6) 1383 FNPVYP(1) QFNPVY(1) FNPAYY(1) MNPAYS(1) MNPVYS(1) MFNPVS(4) YFNPAY(1) AYNPIY(1) YNWVYD(1) FNPVYP 9 Phage display (common panning) PMID:7588601 Anti-fiber tail monoclonal antibody 4D2.5 Tail domain of fiber protein Not determined. Not determined. f3-6mer phage display library (X6) A similar peptide motif, FNPVYP, is present between residues 11 and 16 in Ad2 and was found to be highly conserved within the fibers of other serotypes. 1384 RQFQSP(1) RQNQSP(1) SQFQSP(1) SQFQAP(2) IQFQGP(1) FQWQGP(1) QWQAPA(2) QGQAPV(1) R-Q-[WF]-Q-[SA]-P 8 Phage display (common panning) PMID:7588601 Anti-penton base monoclonal antibody 20G1 Penton protein Not determined. Not determined. f3-6mer phage display library (X6) The best fitting peptide sequence in the Ad2 penton base protein was the RQPFQE motif at position 264, in a region presenting a high probability of accessibility and antigenicity. 1385 GLSIWN(8) FESIWP(1) WDGLSP(1) QSLWVP(1) NGSYPL(8) AQQHYA(2) AQQIML(1) NDMISR(3) IDLMSI(1) RTKMSL(1) HMTRND(1) TNSLRA(1) LTARSI(1) ATSHRI(1) TERLHL(1) 15 Phage display (common panning) PMID:7588601 Penton capsomer Not determined. Not determined. Not determined. f3-6mer phage display library (X6) Ad2 penton was immobilized by adsorption to pre-coated penton base antibody. 1386 NGSYPL(11) GLSIWN(5) VEPATP(2) RWYRPI(2) LGEWSF(1) GRLWDT(1) WDAVNL(1) WKWVNL(1) HSWLSH(1) ASPWLV(1) APWKVW(1) ASQSHF(1) AQFQHH(1) GITHVA(1) QLPAKQ(1) HQLDSA(1) MNTLRY(1) RMNTKH(1) NRMRYL(1) NSQRYK(1) AEKLRY(1) YRSKSS(1) YVKSRF(1) FYTLHT(1) MTSDDL(1) SMDADD(1) DDTYLF(1) LDLDPG(1) YIRVEP(1) RDPTYE(1) RQDPRS(1) RWYSPI(1) RTYRPI(1) RAAWPI(1) KAIWRW(1) 35 Phage display (common panning) PMID:7588601 Penton protein Fiber protein 2C9F,1X9T,4V4U, Not determined. f3-6mer phage display library (X6) 1387 GLSIWN(6) NGSYPL(12) GQQDYA(3) 3 Phage display (competitive panning) 3 PMID:7588601 Penton protein Fiber protein 2C9F,1X9T,4V4U, Not determined. f3-6mer phage display library (X6) Elution was carried out by competition with the natural ligand of penton base, soluble wild-type full-length fiber (fl-fiber). 1388 GRIFRH(2) GLSIWN(1) MVLFWS(1) TLSVFG(1) KWDTRR(1) DRIVWW(1) AQQHYA(1) NTQYQL(1) QGHYVA(1) NDMISR(1) VLENSM(1) FSTNGM(1) IVGMLR(1) MPKSLR(1) PQLVPS(1) IGLPTR(1) RARRSI(1) RIRLEN(1) KKCCYI(1) TRLYDR(1) DRRYMP(1) 21 Phage display (common panning) PMID:7588601 Fiber protein Penton protein 2C9F,1X9T,4V4U, Not determined. f3-6mer phage display library (X6) 1389 KKCCYI(2) TRVSWE(1) SVAISW(1) IVLWSF(1) TIFITI(1) PKLGNH(1) PVRHPN(1) MPAHPF(1) PHFARS(1) SPRNKF(1) RTPTLF(1) LPRARA(1) RIRSLI(1) STLVRY(1) YRVLRT(1) THSKSF(1) SHHSSQ(1) GAFFIG(1) GAILVI(1) KNLFFI(1) SSRVFF(1) LSAGLA(1) 22 Phage display (common panning) PMID:7588601 Head domain of fiber protein Penton protein 2C9F,1X9T,4V4U, Not determined. f3-6mer phage display library (X6) 1390 KCCYST(2) IRVRTW(1) LVRQVF(1) MRLTNM(1) GLRMFM(1) KKCCYI(1) KCCFYV(1) AGIFRH(1) VRSHLL(1) HKTLGF(1) TEHRID(1) VLTVIH(1) ITRSSG(1) LAGTRL(1) NEFSNY(1) ELASVY(1) DARELY(1) GGFELI(1) GALLFP(1) 19 Phage display (common panning) PMID:7588601 Shaft domain of fiber protein Penton protein 2C9F,1X9T,4V4U, Not determined. f3-6mer phage display library (X6) 1391 RRVLLG(9) RYVLPI(1) 2 Phage display (competitive panning) 3 PMID:7588601 Fiber protein Penton protein 2C9F,1X9T,4V4U, Not determined. f3-6mer phage display library (X6) Elution was carried out by competition with the natural ligand of fiber, soluble penton base (pb). 1392 GHSSLGISRWVG(1) GNAQRIFSPRSY(1) AAFGSNRVELFM(1) GLLSIIWNSSAR(1) RGAEWYMVNEGD(1) MNLARPNATSDM(1) 6 Phage display (common panning) 6 PMID:8612228 PEA10 mouse fibroblast cells Not determined. Not determined. Not determined. Phage display library ON159.3 (X12) Binding was conducted at 4 ℃ and the phage were eluted from the cell surface with acid. 1393 SALNPWDEYLEL(3) IASVVRTEVAGF(1) 2 Phage display (common panning) 6 PMID:8612228 PEA10 mouse fibroblast cells Not determined. Not determined. Not determined. Phage display library ON159.3 (X12) Binding was conducted at 4 ℃. The cell were incubated with acid, then scraped from the plate and spun in a vortex mixer. This cell-associated fraction was recovered and amplified. 1394 TPHSLYEDLKRQMMQLGRHL(5) 1 Phage display (common panning) 6 PMID:8612228 PEA10 mouse fibroblast cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Binding was conducted at 4 ℃. The cell were incubated with acid, then scraped from the plate and spun in a vortex mixer. This cell-associated fraction was recovered and amplified. 1395 MKVPDPVSNASN(1) WNYTISERATRD(1) NYTSTIMHRGYR(1) SGNGSYFWVFPL(1) GMRQAPDFAV(1) 5 Phage display (common panning) 6 PMID:8612228 PEA10 mouse fibroblast cells Not determined. Not determined. Not determined. Phage display library ON159.3 (X12) Binding was conducted at 37 ℃. The cell were incubated with acid, then scraped from the plate and spun in a vortex mixer. This cell-associated fraction was recovered and amplified. 1396 TPHSLYEDLKRQMMQLGRHL(1) KTLTLEAALRNAWLREVGLK(3) 2 Phage display (common panning) 3 PMID:8612228 PEA10 mouse fibroblast cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Binding was conducted at 37 ℃. The cell were incubated with acid, then scraped from the plate and spun in a vortex mixer. This cell-associated fraction was recovered and amplified. 1397 KTLTLEAALRNAWLREVGLK(5) 1 Phage display (common panning) 6 PMID:8612228 PEA10 mouse fibroblast cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Binding was conducted at 37 ℃. The cell were incubated with acid, then scraped from the plate and spun in a vortex mixer. This cell-associated fraction was recovered and amplified. 1398 WSLFLNHAE(12) PWLKYAHEA(1) YTWRWDSKL(1) MRIIHIAVL(1) KWLTTHDGT(1) RFLPYYEIP(1) LNHGHRQLV(2) KFLALMPSQ(1) WKWRYFSSQ(1) VNWHWTVTV(1) RWISIKEHA(1) WWWQTFDAR(1) 12 Phage display (common panning) 3 PMID:11457994 Anti-DENV1 envelope protein monoclonal antibody 4E11 Envelope protein Not determined. Not determined. X9 phage display library 1399 TCTSDILPTPHPCT MCNTDASYPHVPCD KCNSAVVPLGAQCR ACQGKPNLPQNTCP VCTPLKKVVDHPCP RCYNPAEWAEPLCY QCYPHVSTLGWSCE 7 Phage display (common panning) 2 PMID:18760481 Anti-WNV E protein monoclonal antibody E24 Envelope protein Not determined. Not determined. XC(X)10CX phage display library 1400 ACPSFIQALTPECM RCVDSFVINNHMCD QCDSFVISPPRTCG QCNSFAIPPSLLCT HCDSFWIPPPPVCK YCDAFLIEQANHCW KCMDAFVIYETNCQ RCDLRYDAFVPWCP DCFNSKNWEHAQCT SCDVFLTMFYPGCK NCKSVRSGVIERCW SCNSFMIPAQPSCT HCPQDPRSYNAFCI QCDAFAVPVEGACM 14 Phage display (common panning) 2 PMID:18760481 Anti-WNV E protein monoclonal antibody E60 Envelope protein Not determined. Not determined. XC(X)10CX phage display library 1401 ICEDDEEWYSAMCA QCNAREWSANYICR VCDQVQWMTRLACQ TCLEVNWLTRMACM MCEYSHWSGFAFCK TCKELQWLPRLRCL TCSPGLREWAHNCL RCEQSMWTQRLPCL ECEHWGGQSQVQCR ECEWTHNRYTHKCQ 10 Phage display (common panning) 2 PMID:18760481 Anti-WNV E protein monoclonal antibody E113 Envelope protein Not determined. Not determined. XC(X)10CX phage display library 1402 DCEIPKIPMDRKCW YCRTNQSGAFARCT LCAPCAVPGNEAWS LCEETPPNETTPCI HCRMYEKLHHTACL LCKTEARTRAAPCA QCETESDLLIPSCP HCGQNSHPMWRKCQ YCGELAHHKTFMCN RCSCRPPSEATTCK NCGSRTQYCKSRCT TCYSFPMQRAAACP GCKLAMTTTTLACH ACTSQLLSVPSCCC RCPYYTRHPSMRCN GCEMSCEIALVVCH ECPQSTSIGLFRCT FCLPTRESTGRACP ICTSHQHILKPMCE ACDPSTVMVPWPCS YCTPSDRFIPHPCS 21 Phage display (common panning) 2 PMID:18760481 Anti-WNV E protein monoclonal antibody E121 Envelope protein Not determined. Not determined. XC(X)10CX phage display library 1403 LCEIKRVYLRQECM ECTVGYRQQKQECM ACRRQLTDAQTTCL YCIMSEEKPMPRCT KCPKAQKDKELSCQ RCPERHGHHTPACP PCLGAHVRFGNECL QCEREGVKAICTCP VCAVFVVLRTAPCQ PCKLPEMQTEATCL IWSPSPVPPHDECK CCEAEVRHPQLQCV KCSHLAPEATLECN KCSRPARARPTNCE LCPESRVHSRTVCP KCAHRTQPPACMCE SCEMSQQVLIRSCS ACVHSTHRLQENCP PCMGESAETTSKCP TCAGTGEICTSDCE VCTNTLAWSAVLSC GCGTRKRTESQTCK PCGHSMNQHEGSCT FCNPRLTTRPPHCR PCDAPVPEYPTDCG SCNQIRQIRESPCL RCERPNTTGTFSCD ICERVEYQVCSYCR RCGEQWPTKEAPCK NCNLLRPYSFSACS KCPPLSHPATSTCT TCPSPPPMMSTTCL TCAESYPPKMAGCL TCKQNRSWRAKECP LCELGAESREPECK KCAHQNSTTQNMCH ACQSTATQKEPTCR HCASTPQPYEPPCE ACPARLSRPASICP RCESNGPQCTEACR FCEPLWRQDRTLCH RCEPHELESPIGCR ACWQRESQTKDNCC SCLTLRTPATESCA RCRTERDSMCERCR ICNLDQQTSVRDCP TCTQTSQQGSMMCR QCPMLHKVTPLECV QCPNWYSIKTGACV SCAPTLNRHAHECL QCCSEQAGPRIACT TCQMRPRSLFSPCG TCYSMHTMSYGLCA TCYVNILTQPEICA NCRKMEPPAETECA FCETTSYTPPGECL LCAYPAPSKEMSCP TCTSAPHNSREMCK LCIPNPEMRQAPCW QCEHMEFKGHNKCH SCLQEELAAEKYCL PCQRMRAENTQRCA ACEESRTRPASTCR HCNQLSSFAAKKCN HCSCLLSAPKPACE QCQSIPFRTPEACP RCTTRRPRLQDFCE PCPHSMEAHADLCH YCAMLSPTPELRCT RCKEAHPTEPSLVG HCRTSGGETATACL QCAEKVPFCAKHCA LCPETPLCTKSICP ECYHWYPPEGKQCP QCRWIQSGTKKNCY ACIQEIRPPSSQCK SCPGLGQRKGAHCE VCENKNQAQKPWCA ECTYRTRAMPTTCS ECSPAASMKSTSCG ICHYEVPRSLTTCR VCPSTYLKNENHCF ECDQRPTVAPGFCK LCTDEKSALVSTCP SCHEQQRRPVAWCQ KCMMLELISTKACG ECPGSQPQCQSRCT DCAMMNLRTPPHCQ VCQNLRNRLPMECF ECIRPKQPNWTMCW QCGHCDDMETSECL PCTASQTPHGSECI HCHPSTAPEIRSCS ACTVPVTMWNNLCR WCLPLGVTPRDLCQ ACRTKEMPGKNPCS NCPNPKLPHWAPCP NCEWATHAATVTCA ECNSNEPGTPSACV ECPLRPPESSSTCA DCLTPGPCETNYCP TCKTETDLSMYQCR GCKTPQESNNTACH RCEPHELESPIGCL LCHKHTHECRTHCR TCLEPSSSSLAECK VCEEHEPRDRTQCA LCMQSELITDTPCP NCVQPELVTPPTCL 109 Phage display (subtractive panning) 3 PMID:19038455 Anti-WNV polyclonal antibody West Nile virus, WNV Not determined. Not determined. XC(X)10CX phage display library The phage library was first depleted of phage particles reactive to normal human serum (pooled from five non-infected individuals) by adsorbing the phage library to normal human antibodies bound to Protein G-sepharose. 1404 CRTCAHPGEHA CGPFYLSAPQC CGPFFLSPTSC CGPFFLAASVC 4 Phage display (subtractive panning) PMID:7514533 Anti-HBsAg polyclonal antibody Hepatitis B surface antigen Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) Twelve serum-coated beads were pre-incubated with an excess of M13K07-UV killed phage. 1405 TENFNMWKNDMVEQMH RSNFEAWKWDEAQQGGS RENQTRWYRDMGGDMSS ASYLQWVRDTWVSL ASYWKDWLHDTAGGGVW ADYWRGWKERYNRSL EANYNAWREWVRSDWVS REYVYLWWIDHKTSL 8 Phage display (common panning) 3 PMID:8798975 Anti-gp120 monoclonal antibody GV4D3 Surface protein gp120, SU Not determined. Not determined. X20 phage display library 1406 QMHEDIISLWDQSLKPC ASIVSLWDSL DSRSDIWSIWSLGRERS PKRRSASELWGPSL SAHSPIEGLWAGESL GWREGVVGLWDRHSL YPRETAIQLWSL RGSVSIRDLWRSL 8 Phage display (common panning) 3 PMID:8798975 Anti-gp120 monoclonal antibody GV1A8 Surface protein gp120, SU Not determined. Not determined. X20 phage display library 1407 IHYCAPAGFAILKCNNK SGQREHVGFAILSL TRGGAAAGFAVHGSLSL VVARDHTGFAVLGTSL TDLMHCSGFALSPGCPQ ASMNGKVGWAIHAREAV SDDSSHTGWAVLDLGLR ASKTGVGWAVL QDMVRRPGWALLHPQLG 9 Phage display (common panning) 3 PMID:8798975 Anti-gp120 monoclonal antibody GV4H3 Surface protein gp120, SU Not determined. Not determined. X20 phage display library 1408 KVVKIEPLGVAPTKAKR GGLQRAALGVAPRT LSSKEGALGVAPPLTRT GGGLIKVLGVAPEGDSG PGALGVAPRVMER AGRAEWPLGIAPG ARGVAPDPTEH RVRKVEPLGVRVTA KVGKRSPLGIGPHAGG 9 Phage display (common panning) 3 PMID:8798975 Anti-gp120 monoclonal antibody GV1G2 Surface protein gp120, SU Not determined. Not determined. X20 phage display library 1409 XNNKFTSPPMSL DHGASSFHVSSX SDLNTDPSHPST LPLVVTPTSANG LKPISSHPSTVA HDQSPYXTVYGN SYQRAHHPPVLR STSQPTVSTLRP QHKTQPLNVISF TTTLSLVPLRPN GYSLYPIITSQK HTPHPMNLVAEA QTPPMQSPSATE SYSSPPSKAVRE HAPSYPLETKLM FYDSQPQVLSTT MPLMPTARSSMS GPRPPAALPHPL QSSYQTPXTPQP YXTALLHTPAED HTPHPDASIQGV YSNASARELGAT 22 Phage display (common panning) 3 PMID:9878399 Paclitaxel (Taxol) Apoptosis regulator Bcl-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) Thirty micrograms of C7-biotinylated paclitaxel solubilized in HPLC-grade dimethylsulfoxide and diluted into Trisbuffered saline plus Tween 20, was attached to a streptavidin-coated petri dish. Each selection was performed on the streptavidin-coated petri dish. A subset of the peptides was identified that exhibits significant similarity to a non-conserved region of the anti-apoptotic human protein Bcl-2: ELISA assays conffirmed binding of paclitaxel to Bcl-2, and circular dichroism spectroscopy demonstrated that a substantial conformational change accompanies this binding. In vivo, treatment with paclitaxel has been shown to lead to Bcl-2 inactivation with concomitant phosphorylation of residues in a disordered, regulatory loop region of the protein. Similarity between paclitaxel-selected peptides and this loop region implicate these residues in drug binding, and suggest that the apoptotic action of paclitaxel may involve the binding of paclitaxel to Bcl-2. 1410 CKSSGKLISLC[14.98 ± 2.51, 11.73 ± 1.78] CGTKLVCFAAC[12.72 ± 2.11, 14.09 ± 2.18] CCNGRLYCGPC[13.87 ± 2.07, 1.02 ± 0.00] CCAGGLTCSVC[13.73 ± 2.64, 12.50 ± 1.30] CSGRLYCHESWC[18.74 ± 1.30, 10.23 ± 1.30] 5 Phage display (subtractive panning) PMID:10229859 Anti-HIV-1 polyclonal antibody Human immunodeficiency virus type 1, HIV-1 Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA Comparison of ELISA reactivities of phagotopes with sera of LTNP vs AIDS subjects was performed by ELISA. The difference between OD405 nm and OD620 nm was determined by an ELISA reader. Data are expressed as fold increase of the average values of the phagotopes over values of the wild-type phages and as mean ± SEM of fold increase for each phagotope. Data shown are reproduced from Fig. 1B in the reference. Data in the first column are values of phagotopes with sera of LTNP. Data in the second column are values of phagotopes with sera of LTNP. Values were considered positive when at least 4-fold higher than the background signal of wild-type phage. In the immunoaffinity selection, serum IgG was linked to magnetic microbeads previously coated with an anti-human (Fc-specific) polyclonal Ab of beads suspension. Given the fact that sera from long-term nonprogressor (LTNP) subjects show higher titers of neutralizing Abs than sera from AIDS patients, initial screening was performed with LTNP sera. 1411 EATFVYPAP[12.23 ± 1.74, 12.02 ± 2.08] TKTLIYGGA[13.73 ± 2.41, 11.48 ± 1.94] KRIVIGPQT[17.49 ± 1.83, 9.55 ± 1.93] FASLHYDKP[13.92 ± 2.17, 10.52 ± 1.40] RPTLRFQGA[11.56 ± 1.88, 13.32 ± 1.16] 5 Phage display (subtractive panning) PMID:10229859 Anti-HIV-1 polyclonal antibody Human immunodeficiency virus type 1, HIV-1 Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA Comparison of ELISA reactivities of phagotopes with sera of LTNP vs AIDS subjects was performed by ELISA. The difference between OD405 nm and OD620 nm was determined by an ELISA reader. Data are expressed as fold increase of the average values of the phagotopes over values of the wild-type phages and as mean ± SEM of fold increase for each phagotope. Data shown are reproduced from Fig. 1B in the reference. Data in the first column are values of phagotopes with sera of LTNP. Data in the second column are values of phagotopes with sera of LTNP. Values were considered positive when at least 4-fold higher than the background signal of wild-type phage. In the immunoaffinity selection, serum IgG was linked to magnetic microbeads previously coated with an anti-human (Fc-specific) polyclonal Ab of beads suspension. Given the fact that sera from long-term nonprogressor (LTNP) subjects show higher titers of neutralizing Abs than sera from AIDS patients, initial screening was performed with LTNP sera. 1412 NTAHSDMHSP(2) KAEERFHEVR(1) WNIFQSLSSP(1) RVAATAAREE(1) TPEPLEEAAS(1) DHVFKRPQPS(1) ETCVEKNEAD(1) WRRDGC(1) YMDPHTQREA(1) EPVRDNCAPS(1) VERDIATRPW(1) ERPEIEDVCQ(1) QKTLNTSNAN(1) VKTLNTSNAN(1) RDTTMWEVNA(1) WGHCSQGMIE(1) PPFDVFHNPM(1) DTHAGMHSPT(1) RMPHHDPQLM(1) TQNLMQMQHA(1) SPFMLMHGEH(1) TCQAGRESMHNP(1) WETMHNPGTP(1) DALNMHEGRP(1) SKLETTMHSP(1) LGGMDSMHSP(1) MAQMHEPVRS(1) LPQHNMMHDP(1) PSNYSAMHAP(1) RCETHFNMHEPY(1) 30 Phage display (subtractive panning) 2 PMID:19390580 Anti-HER-2 extracellular domain polyclonal antibody Extracellular domain of HER-2 Not determined. Not determined. XC(X)10CX phage display library The library was initially depleted of phages recognized by naive mouse serum by 3 sequential pannings of the library with immobilized serum of non-immunized mice. The resultant \"depleted\" library was used for mimotope identification. Similarly, the sera obtained from immunized mice were initially depleted of anti-phage activity by overnight incubation at 4 ℃ with wild type phage immobilized on plastic plates. The sera were also depleted of IgM by similar incubation with immobilized anti-IgM antibody. 1413 NWSIAGD(1) SRGDTEP(1) TSDSWQR(1) PKTEVPQ(1) DDFSPPR(1) EWYTPQG(1) SRCDLDEKGC(1) PRCKHNQKKC(1) AGACKRMREF(1) ATPKRTHDHD(1) TSNNRVEARA(1) RTTPSGGGFK(1) DGWYVAQ(1) GGRWGES(1) WYTTPGS(1) SGWYTPV(1) AKDEQPM(1) GWYVSSP(1) PSEGQSE(1) GTAGGEITEH(1) FYSSMFWAVGEQ(1) LQEFPGDQLV(1) LSTRLWIPAW(1) KTTTWPSTPT(1) MWASVNKMA(1) PPPLLAGDPK(1) AVNQCTTVLA(1) GTAGGEITEHE(1) 28 Phage display (subtractive panning) 2 PMID:19390580 Anti-HER-2 polyclonal antibody Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. XC(X)10CX phage display library The library was initially depleted of phages recognized by naive mouse serum by 3 sequential pannings of the library with immobilized serum of non-immunized mice. The resultant \"depleted\" library was used for mimotope identification. Similarly, the sera obtained from immunized mice were initially depleted of anti-phage activity by overnight incubation at 4 ℃ with wild type phage immobilized on plastic plates. The sera were also depleted of IgM by similar incubation with immobilized anti-IgM antibody. 1414 TYSGRAACGV(1) SASMPFPANE(1) SKPKYEKPSQ(1) EVTTEHVEAN(1) RSHERHPPSP(1) DDELHSGTSY(1) SPPAFSHPMQ(1) VNTQGPNSIA(1) 8 Phage display (subtractive panning) 2 PMID:19390580 Anti-HER-2 polyclonal antibody Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. XC(X)10CX phage display library The library was initially depleted of phages recognized by naive mouse serum by 3 sequential pannings of the library with immobilized serum of non-immunized mice. The resultant \"depleted\" library was used for mimotope identification. Similarly, the sera obtained from immunized mice were initially depleted of anti-phage activity by overnight incubation at 4 ℃ with wild type phage immobilized on plastic plates. The sera were also depleted of IgM by similar incubation with immobilized anti-IgM antibody. 1415 ADTVSGAMV(7) VARVGSPPD(7) VAVVGTFPD(5) VSTPGSPDD(5) ASYQDLGSS(4) AEMVKSSID(4) DFVTRSPQD(4) EWDPAPAME(4) ARGGEGTED(3) DNGSLANGE(3) DRRDDAVSE(3) ENARSVNDD(3) EYSAPSADA(3) GDYQSLVED(3) GMNNASFPD(3) GSPSEYSSQ(3) VPQTSLAME(3) EGGGGLIND(2) ESGGSGSTE(2) GETLDPSLE(2) GGTESMTAS(2) VFLGDPSVE(2) VSATLEPVD(2) ADGSPDLPA(1) AEGGLEHLD(1) AEYAFGGTD(1) AFGAQEDNT(1) AFNATASMS(1) AGSNINLVD(1) AHMSEVEGG(1) AIRDIAGDS(1) AITASDSMS(1) ANRVSEMTE(1) ANSISFDDD(1) APDSSAWSD(1) APNLSISDS(1) APTVEADNN(1) AQDLAVITG(1) AQYETSYPE(1) ASSHFDGSD(1) AYESGTGSS(1) DEHMEGNAG(1) DGFADLASD(1) DGRPSDIPT(1) DGVADAASN(1) DSGGSGIAS(1) DVSGAFMAD(1) EAGPIPGDG(1) EDISLSMGE(1) EDYPDISVG(1) EETMNGMPM(2) EFNDGSGVD(1) EHELDPVMA(1) EIGGMGAGG(1) EMERSSSMA(1) ENPVLASDS(1) ESSTQDFPD(1) EWVDPRGED(1) EYSSSELPP(1) GATHEVGGD(1) GDLVQDVTA(1) GESSGGADD(1) GGDFAAANG(1) GLRGEVEEG(1) GMQAGDQDD(1) GPVEGDADG(1) GSDSDLTFD(1) GSGGAENVS(1) GSLDDITDN(1) GSRDGVISE(1) VEGQGSVSD(1) VGDAGHSME(1) VGEGYREES(1) VGRDSTLND(1) VGTDSNDAG(1) VLDSVSSPD(1) VLPVGAGTD(1) VNENQLVMD(1) VNVTDAGID(1) VPAAYIGND(1) VTANPDSSA(1) VTMFESNAD(1) VTNDPSLDD(1) VVNQGDPPD(1) VVRDPGVSG(1) VYQNSESTL(1) 86 Phage display (subtractive panning) 3 PMID:19186939 α, β-tubulins from bovine brain Microtubule-associated proteins (MAPs) Not determined. Not determined. f8-9mer phage display library (X9) The landscape phage library was added to the untreated dish at 4 °C to remove the phages that specifically bound to the Petri dish. The tubulins received have been modified so that random surface lysines contain a covalently linked, long-chain biotin derivative. This long-chain biotin derivative enables the immobilization of the tubulins on the streptavidin-coated Petri dish. By using RELIC, the affinity-selected peptides were shown to have similarity with the sequences of two MAP families (MAP1 and MAP2/tau), thereby identifying putative microtubule-binding domains on these MAPs. The tubulin-binding affinity was also confirmed by using transmission electron microscopy (TEM) to characterize the interaction between affinity-selected tubulin-binding phage and tubulins. 1416 YTRSEGVLLQLT(1)[++] FGCRVESVSTHC(1)[+++] CRNQAVRVFCHN(1)[+++] EKPSESRIAQLG(1)[++] KGFRMEAVWRRF(1)[+] YVWKIVTEGVNV(1)[+] RKRCFHALDCLG(1)[+] R-x-[EQ] 7 Phage display (subtractive panning) 2 PMID:18707547 Anti-CD6 monoclonal antibody ior t1 T-cell differentiation antigen CD6 Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA The affinities of the selected clones to anti-CD6 monoclonal antibody ior t1 were analyzed by ELISA assay. The absorbance was measured at 492 nm. Symbols represent the optical density values obtained for each antibody as follows: (+) from 0.3 to 0.5; (++) from 0.6 to 1.0; (+++) from 1.1–2.0. Polystyrene beads were preincubated with approximately M13K07 UV-killed phage particles. 1417 KGIPNTKAP[0.589] DMFQIGKYG[0.405] GIREVWPAG[0.917] SSMGAYWGG[0.241] KGTTGVRNT[0.441] 5 Phage display (common panning) 3 PMID:17964653 Anti-Der p 1 monoclonal antibody Peptidase 1 Not determined. Not determined. J404 phage display library (X9) ELISA Der p 1-specific was coated on ELISA plates and incubated with selected single phage clone amplificates. The detection was carried out with anti-phage antibody at 405-490 nm. For negative control wild-type phage (wt) lacking the specific insert was used at the same concentration as the selected phage clones. Absorbances at 405-490 nm were reproduced from the graph and shown. The absorbance for negative control was 0.064. To identify single phage clones, 120 random phage clones were screened by a colony screening ELISA. On this ELISA, five phage clones showed a significantly higher binding signal to the specific antibody in comparison to background reactivity with wild-type phage. No binding of these clones was detected to the control antibody. 1418 FVVEYTKKW[0.229] SWWNLPQIG[0.358] KGITTKWMA[0.419] AGISYTKTW[0.702] 4 Phage display (common panning) 3 PMID:17964653 Anti-Der p 2 monoclonal antibody Mite group 2 allergen Der p 2 Not determined. Not determined. J404 phage display library (X9) ELISA Der p 2-specific was coated on ELISA plates and incubated with selected single phage clone amplificates. The detection was carried out with anti-phage antibody at 405-490 nm. For negative control wild-type phage (wt) lacking the specific insert was used at the same concentration as the selected phage clones. Absorbances at 405-490 nm were reproduced from the graph and shown. The absorbance for negative control was 0.067. To identify single phage clones, 120 random phage clones were screened by a colony screening ELISA. On this ELISA, four phage clones showed a significantly higher binding signal to the specific antibody in comparison to background reactivity with wild-type phage. No binding of these clones was detected to the control antibody. 1419 PSYNASLDCPYSVGD FSYRSSYVPSSRRSA RCEVWPCRSSPHLNP CGVGHCLPFFCFPSF DYFVTPCCWVSRLTL CRRVVCFCDGYCLAD FYSCMFPYAASASFV TLLFVSVPACFGPHA PLSEIFYFGCCYDSV SGVTCCDSSRALCGS GLSVMCNPVLDPVCK PMGYFSFPWNTDVVA FIRHSITLKCAARGG YLFLNVIGSDSFVFD LVDNCILTVFPPHNA LDMFSFPSVCFGTCI RGVSLCHASASSTQY SFYRSDGVLCPIVPW LGNLCHFNAMGLADG VCGLVHVKSLAPPAW VYDASVFGVWTPLHG WATVVADALHLGGFS CCFSGSHLVPYAKGT FFSLTGVPGNVFCLS CVFDVRSAADSFLAV CGNCCVYLDADNDVP SIALGCRGFWYFDLP TVLFALMRSGADGIW FCISHILNYPFDGDA AFLSFSRPLWPSFFC VCLGVGVGFVADCPD LVGALGVTVRHALGT LFHLYCDFDAPLAAW LMICAGRWRSGSFVD CVFFISLNGAACCVG 35 Phage display (common panning) 1 PMID:18930404 Synthetic ligand for FK506-binding protein, SLF Not determined. Not determined. Not determined. X15 T7 phage display library The screening procedure involved a cuvette type 27-MHz quartz-crystal microbalance (QCM) apparatus with introduction of self-assembled monolayer (SAM) for a specific small-molecule immobilization on the gold electrode surface of a sensor chip. For SLF-bio immobilization, the gold sensor surface was initially coated by treatment with 100μl of neutravidin solution (100lg/mlaq) for 30 min at room temperature. After washing the gold electrode surface, SLF-bio was applied and left for 16 h under a humid and shaded atmosphere at room temperature. The gold surface was thoroughly washed prior to conducting the screening experiments. 1420 CDRRACRVAFSSRTG LVFSLFVILLLVLGV FGFFSSNPLVHVHWD FIPAYCCVVCAYLSV GYFNCLVVHRHALRC LSSLCGLPFGRPQCV SFARLLFRLCLTTTP FFFGGLRLLVLLRES FFYRLVFCHYPRLVF DCSRFSWLSSVFAVA HLYGFSLIMFCMVTI SLVLFTSNVYPMFVF WDSSCFVPFLCSGSS CSDIVVASPSPFPTC CVLSLCLFMVLFFSV FFLPVGFRAFLLGRR FVDSCLLCTFTTLRT LFVLSSSDAHECSAY RCVLSYGPNVFQPVD SFCISCLCVCGGDGR SFDAALCLVSSLFGA TGSHCGVYKVGLGVV VTYFTWSFDPRGCCW VVVIVFLCLVWLVGG YVLWWRLLFFILIIS YYAIFFHRVLRMYSV SFDFGSFDFVSPLAP CIDVNCFVWTFGMPF FGYELSFLSSWSVFG 29 Phage display (common panning) 1 PMID:18930404 Irinotecan, Iri Not determined. Not determined. Not determined. X15 T7 phage display library The screening procedure involved a cuvette type 27-MHz quartz-crystal microbalance (QCM) apparatus with introduction of self-assembled monolayer (SAM) for a specific small-molecule immobilization on the gold electrode surface of a sensor chip. For Iri-bio immobilization, the gold sensor surface was initially coated by treatment with 100μl of neutravidin solution (100lg/mlaq) for 30 min at room temperature. After washing the gold electrode surface, Iri-bio was applied and left for 16 h under a humid and shaded atmosphere at room temperature. The gold surface was thoroughly washed prior to conducting the screening experiments. 1421 PLDMIGTYQHIM(60) 1 Phage display (common panning) 3 PMID:18383103 Anti-transferrin monoclonal antibody hTf345 Serotransferrin, Transferrin Not determined. Not determined. X12 phage display library 1422 EPVESPWSRARL(2) VVGTDPFSRARL(4) AGPRDPYGV(1) QGMKSPYDSARL(1) LEYDPHHVARV(1) TLQTTPHDVARL(1) KKFKMGPPV(1) ETPTPWGRARV(4) TLQTTPHDVARR(4) SQGKTPFDIGRR(1) DPHWVGRV(1) GDPHYVARV(1) 12 Phage display (common panning) 3 PMID:17397983 Anti-b558 monoclonal antibody 7A2 Cytochrome b-245 heavy chain Not determined. Not determined. J404 phage display library (X9) 1423 SCTDDPWCTARV(9) HERGDPHYVARV(1) RIVQDPVDVARR(1) DRERDPWLVARR(1) RWNHTSERV(1) 5 Phage display (common panning) 3 PMID:17397983 Anti-b558 monoclonal antibody 8G11 Cytochrome b-245 heavy chain Not determined. Not determined. J404 phage display library (X9) 1424 DHQRFFV(5) AHQASFV(1) 2 Phage display (common panning) 3 PMID:11292756 Anti-E. coli K1 monoclonal antibody HmenB1 Escherichia coli K1 bacteria Not determined. Not determined. Ph.D.-7 phage display library (X7) The new mimotopes may be useful in producing a vaccine(s) capable of eliciting anti-NMGB antibodies not reactive with neuronal tissue. 1425 SHVPNAF(23) SHAPNAF(6) SHVPDAF(4) SHVHNAF(2) SHAPSAF(1) SHVPHAF(1) 6 Phage display (common panning) 3 PMID:11292756 Anti-E. coli K1 monoclonal antibody HmenB3 Escherichia coli K1 bacteria Not determined. Not determined. Ph.D.-7 phage display library (X7) The new mimotopes may be useful in producing a vaccine(s) capable of eliciting anti-NMGB antibodies not reactive with neuronal tissue. 1426 CTGPGSWFC(6) CTSPGPWFC(2) CTAPGAWFC(1) CTTPGPWFC(1) 4 Phage display (common panning) 3 PMID:11292756 Anti-E. coli K1 monoclonal antibody HmenB3 Escherichia coli K1 bacteria Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The new mimotopes may be useful in producing a vaccine(s) capable of eliciting anti-NMGB antibodies not reactive with neuronal tissue. 1427 VVSTGSH(5) IPMKGHW(1) LPMKYQA(1) HWGMWSY(1) ALSSDPH(1) DPRLSAL(1) KPSTLML(1) YHWYTSP(1) LFMPATP(1) ALLPWTD(1) SPPPPPI(1) SHAPYTH(1) NQDVPLF(1) 13 Phage display (common panning) 3 PMID:11292756 Anti-E. coli K1 monoclonal antibody HmenB13 Escherichia coli K1 bacteria Not determined. Not determined. Ph.D.-7 phage display library (X7) The new mimotopes may be useful in producing a vaccine(s) capable of eliciting anti-NMGB antibodies not reactive with neuronal tissue. 1428 MELQTRS(5) LPMKYQA(5) NTLQPSP(2) YMTPFSP(2) LHAQSRS(1) YHLQPTP(1) HFMEPVN(1) LSTPSLL(1) ITAARLP(1) SIDNPLP(1) SIITGYL(1) AYSMLGH(1) 12 Phage display (common panning) 3 PMID:11292756 Anti-E. coli K1 monoclonal antibody HmenB14 Escherichia coli K1 bacteria Not determined. Not determined. Ph.D.-7 phage display library (X7) The new mimotopes may be useful in producing a vaccine(s) capable of eliciting anti-NMGB antibodies not reactive with neuronal tissue. 1429 LEKGTPW AAKGTPW EPDKMTT EPDKSII QKTAASY GKQPASD AVTFSKG 7 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #1 Major pollen allergen Bet v 1-A Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Bet v 1. 1430 LEKGTPW AAKGTPW QKTAASY 3 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #1 Major allergen Pru av 1 Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Pru av 1. 1431 LEKGTPW AAKGTPW QVELPKG KLPLTKG QKTAASY EAAKQGE 6 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #1 Stress-induced protein SAM22 Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Gly m 4. 1432 LEKGTPW AAKGTPW EPDKSII EAGKHSG 4 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #1 Ara h 8 allergen Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Ara h 8. 1433 GKIGMYP GKDRSIQ IKLPALF HLVEKGY 4 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #2 Major pollen allergen Bet v 1-A Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Bet v 1. 1434 GKIGMYP FKLKAIE ITADMYT MPNYDRL 4 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #2 Major allergen Pru av 1 Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Pru av 1. 1435 GKIGMYP GKDRSIQ IKLPALF HLVEKGY 4 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #2 Stress-induced protein SAM22 Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Gly m 4. 1436 GKIGMYP IKLPALF 2 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #2 Ara h 8 allergen Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Ara h 8. 1437 KNLEEPK KYLEEPV LNRADIL WPKDTDT 4 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #3 Major pollen allergen Bet v 1-A Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Bet v 1. 1438 KNLEEPK KYLEEPV TPIRHRS QYPWRLA FDMATNP SLYELTH TAIHAVW DHLPSPW 8 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #3 Stress-induced protein SAM22 Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Gly m 4. 1439 YKIGDNW KSDNNVT 2 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #4 Major pollen allergen Bet v 1-A Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Bet v 1. 1440 YKSDNIR SKTDNLT AFNRASD 3 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #4 Stress-induced protein SAM22 Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Gly m 4. 1441 YLMEVQK TLKQAPA 2 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #5 Major pollen allergen Bet v 1-A Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Bet v 1. 1442 YLMEVQK YEMDAPK ERHLTFT 3 Phage display (competitive panning) 2 PMID:16199263 Polyclonal antibody IgE from birch pollen-allergic patient #5 Stress-induced protein SAM22 Not determined. Not determined. Ph.D.-7 phage display library (X7) Two millilitre aliquot of a tosyl-activated M280 Dynabeads suspension were loaded with rabbit-anti-human IgE , and blocked with skim milk in PBS. Patient serum (600 μl) was diluted in 9.4 ml PBS with 0.05% Tween 20 and 0.1% skim milk, and incubated overnight at 4 ℃ with 200 μl coated beads suspension. In the second round, phages bound to the beads were eluted with Gly m 4. 1443 GCLYSDLLATCI[1.321] NCLYSDLTQSCI[1.236] NCLYSDLYARCI[1.223] KCMYSDLLGICI[1.153] DCLYSDLESRCI[0.818] SCLYSDLLELCI[0.750] ECMWSDLELRCI[0.571] NCLWSDLEQFCI[0.343] 8 Phage display (common panning) PMID:11413337 Anti-gp120 monoclonal antibody b12 Surface protein gp120 (SU) 2NY7, 1N0X, XCX(3)SDLX(3)CI phage display library ELISA Affinities of selected phages to IgG1 b12 were measured by ELISA. OD405-490, optical density at 405 minus 490 nm, was shown. The absorbances of phage gp120 as positive control and phage f88-4 as negative control were 1.121 and 0.075, respectively. 1444 HERSYMFSDLENRCI[1.236] CSRNQLWSDLHGSCI[1.207] NNQGCLWSDLTASCI[1.189] STTRCTWSDLYDSCI[1.141] QSSSCMWSDLFQQCI[0.992] AQKQCTWSDLLSRCI[0.903] RPCRGVYSDLLDKCI[0.886] SSDHCLWSDLTMTCI[0.644] LPSSCSWSDLLNRCI[0.276] HTCAGTWSDLLSTCI[0.252] 10 Phage display (common panning) PMID:11413337 Anti-gp120 monoclonal antibody b12 Surface protein gp120 (SU) 2NY7, 1N0X, X(7)SDLX(3)CI phage display library ELISA Affinities of selected phages to IgG1 b12 were measured by ELISA. OD405-490, optical density at 405 minus 490 nm, was shown. The absorbances of phage gp120 as positive control and phage f88-4 as negative control were 1.121 and 0.075, respectively. 1445 FNWSLDYLREYH(6)[0.537] ISTVPNYLRHFH(6)[0.536] DKYRIVIARGCV(5)[0.544] LCHPQNCHTKSL(3)[0.547] DPYKLPNRRCCI(3)[0.603] YTIADLITRCCV(2)[0.598] PRDTNTIRRCCV(2)[0.584] PYYNRHHPQDNR(2)[0.556] WGPYPEWLRFYH(2)[0.552] MPDLTTHPQFAR(1)[0.553] RWIDATSHPQFD(1)[0.533] HHPMDNRSYRWP(1)[0.522] GVLHPFHHPGNV(1)[0.530] VVFRPHYLSGFH(1)[0.557] NYHHLRDLRSYH(1)[0.532] HMASWLRFFHGP(1)[0.529] DDFRFTLPRCCV(1)[0.612] DRILAIGYRCCV(1)[0.561] MHVETALVRCCI(1)[0.543] PDRLRSPMRCCI(1)[0.532] DVGFIDRNWCCV(1)[0.360] H-P-[QM], [YW]-L-R-x-[YF]-H, R-C(2)-[VI] 21 Phage display (common panning) 3 PMID:16819727 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. X12 T7 phage display library ELISA After three rounds of biopanning, approximately 1.0e11 PFU of amplified phage pools isolated from each round of screening were analyzed for SA binding in a polyclonal (heterogeneous phage populations) phage ELISA. Absorbances at 650 nm were determined and shown. The absorbance of the phage T7 10-3b as negative control was 0.050. 1446 CHPMRPC(5)[0.654] CHPQFRC(5)[0.638] CHPQVRC(4)[0.630] CHPQGPC(3)[0.677] CVHRFVC(3)[0.639] CVHRFIC(2)[0.668] CHPMVPC(1)[0.613] CPHPMFC(1)[0.582] CHPQFIC(1)[0.676] CHPQFNC(1)[0.728] CHPQFPC(1)[0.774] CHPQNHC(1)[0.707] CHPQNNC(1)[0.667] CHPQNPC(1)[0.751] CHPQVKC(1)[0.597] CHPQVMC(1)[0.602] CHPQYPC(1)[0.632] CKHPQFC(1)[0.639] CLHPQFC(1)[0.681] CRHPQFC(1)[0.666] CIHRFIC(1)[0.696] CIHRFLC(1)[0.662] H-P-[QM], C-[VI]-H-R-F-[VIL]-C 22 Phage display (common panning) 3 PMID:16819727 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. CX5C T7 phage display library ELISA After three rounds of biopanning, approximately 1.0e11 PFU of amplified phage pools isolated from each round of screening were analyzed for SA binding in a polyclonal (heterogeneous phage populations) phage ELISA. Absorbances at 650 nm were determined and shown. The absorbance of the phage T7 10-3b as negative control was 0.060. 1447 CKRHPQVGC(18)[0.703] CVRHPQFGC(6)[0.702] CRKHPQVGC(5)[0.721] CWQVMHLKC(5)[0.623] CQRHPMVGC(4)[0.728] CGHPQFHFC(3)[0.723] CRQMMHITC(3)[0.615] CLPMMHITC(2)[0.515] CHPMAPLRC(2)[0.721] CGHPQRWTC(2)[0.703] CHPQAPRWC(2)[0.731] CGFRHPQFC(1)[0.755] CGHPQIGRC(1)[0.734] CHPQAGKRC(1)[0.757] CHPQFATRC(1)[0.758] CHPQFWRLC(1)[0.817] CHPQNDTGC(1)[0.635] CHPQVYRAC(1)[0.658] CHTKHPQFC(1)[0.708] CLHPQSGRC(1)[0.661] CRKHPQIGC(1)[0.690] CHPMAARNC(1)[0.723] CHPMAPRTC(1)[0.659] CKRHPMVGC(1)[0.676] CKRHPHVGC(1)[0.722] CLHPMNNRC(1)[0.666] CWAVMHIPC(1)[0.543] CWPVMHAFC(1)[0.681] CFVGYGFLC(1)[0.655] CNAMPWFMC(1)[0.605] H-P-[QM], C-x(2)-[VM]-[M]-H-[LIA]-x-C 30 Phage display (common panning) 3 PMID:16819727 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. CX7C T7 phage display library ELISA After three rounds of biopanning, approximately 1.0e11 PFU of amplified phage pools isolated from each round of screening were analyzed for SA binding in a polyclonal (heterogeneous phage populations) phage ELISA. Absorbances at 650 nm were determined and shown. The absorbance of the phage T7 10-3b as negative control was 0.060. 1448 CDPATPYC(43)[0.850] CDLAAPYC(7)[0.752] CDPPTPYC(3)[0.780] CDLSTPYC(2)[0.675] CDVATPWC(2)[0.769] CDWSSPYC(2)[0.812] CDLATPYC(1)[0.809] CDPASPWC(1)[0.877] C-D-P-A-[TS]-P-[YW]-C 8 Phage display (common panning) 3 PMID:16819727 NeutrAvidin Biotin 1AVD,2C4I,2JGS,1DF8, Not determined. CX6C T7 phage display library ELISA After three rounds of biopanning, approximately 1.0e11 PFU of amplified phage pools isolated from each round of screening were analyzed for NA binding in a polyclonal (heterogeneous phage populations) phage ELISA. Absorbances at 650 nm were determined and shown. The absorbance of the phage T7 10-3b as negative control was 0.061. 1449 AFDVRERLL(3) IDWRSRLAP(3) KHDFRSRLA(3) ADWRERLVL(2) AVDWRQRML(2) KTDFRSQLL(2) QDFRWLLT(2) SMDIRNRLL(2) WMDGRHRLL(2) EFRGRLLET(2) ADYRSRLTW(1) AFHVRDRFL(1) AGDFRSRLF(1) ALDWRLRMG(1) ALEFRGRLL(1) DFRGRLFNL(1) DFRNRLTAP(1) DFRLRLLGS(1) DFRSRLLRV(1) DFRLRLVNP(1) DFRYRLLQS(1) DIDFRSRML(1) DLDWRQRLF(1) DTDFRQRLL(1) DWRHRLVNH(1) DWRSRLVIG(1) DWRGRLS(1) EGGYRQRLL(1) EMGFRARLL(1) FDFRSRLLS(1) GADIRARLL(1) GDWRARLAV(1) GIDYRARLL(1) GKDWRGLLL(1) GLDPRARLL(1) GLDWRISLL(1) GPDSRARLL(1) GQDHRERLL(1) GVDFREKLT(1) GWDNRARLL(1) HLDFRARLL(1) HQDWRANLL(1) HSEWRARLA(1) IDWRARLTK(1) ILKDWRHRL(1) IPSFRDRLL(1) IRDFREMLL(1) KDWRGRLVN(1) KLDSRHRLI(1) KMDWRGRLM(1) KQDIRARLL(1) LDFRGSLLV(1) LDIRGRLLG(1) LDIRHRLVN(1) LEDWRGRLL(1) LIDYRQRLA(1) LLDSRARLI(1) MDPRSRLMT(1) MFDFRERML(1) MHDFRARLL(1) MLDRLERLL(1) MLDPRNRLL(1) MNDPRDRLL(1) NDIRGRLLT(1) QADFRDRLL(1) QDFRNRLQL(1) QEDWRGLLL(1) QKDFRNRLA(1) QSSFRERLL(1) RRMLRARLL(1) SDWRGLLL(1) SDWRGRLGA(1) SELFRSRLL(1) SFDWRGKLL(1) SLWDWRGRL(1) SMDLRSRL(1) STDFRQRLF(1) TDDFRSRPL(1) TDFRHRLGM(1) TIEFRHRLL(1) TRDVRERLL(1) VADIRARLL(1) VGDWRHRLM(1) VHGDFRNRL(1) WDFRNRLV(1) WDLRERLI(1) WFPEWRMTS(1) WGDFRANLI(1) WGGFRDRLL(1) YEDWRGRLL(1) 90 Phage display (common panning) 3 PMID:17675514 Anti-FPR monoclonal antibody NFPR1 fMet-Leu-Phe receptor, fMLP receptor Not determined. Not determined. J404 phage display library (X9) 1450 LPSISDLL(28) ETTLPSIKW(13) GALDQSLPS(2) PSISDLLAI(2) RDTLPSIFW(2) RETLPSLYL(2) SQWWRHNDM(2) TLPSLMRHI(1) TTLPSISDLL(1) TTLPSISDWL(1) WWGGTLPSL(2) AATLPSIDW(1) LPSIDAVLN(1) ESIAFWNRRPA(13) DSIAFWNRRPA(12) CLTCWNVMPA(1) EVASYWRWG(9) ETLDWWRVG(4) ETRDWFRFE(3) EVRGWSRIW(1) EVFGWDRFW(1) EVIGWSRLF(1) ETRDWWRLS(1) KHLHRHKIG(1) LCIPGFC(1) 25 Phage display (common panning) 3 PMID:17675514 Anti-FPR monoclonal antibody NFPR2 fMet-Leu-Phe receptor, fMLP receptor Not determined. Not determined. J404 phage display library (X9) 1451 AYSSGAPPMPPF NPSSLFRYLPSD SLATQPPRTPPV 3 Phage display (common panning) 4 PMID:12618805 Silver nanoparticles Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1452 KFLQFVCLGVGP AVLMQKYHQLGP IRPAIHIIPISH NVIRASPPDTSY LAMPNTQADAPF QQNVPASGTCSI NAMPGMVAWLCR HNTSPSPIILTP ASQTLLLPVPPL YNKDRYEMQAPP TLLLLAFVHTRH PWATAVSGCFAP SPLLYATTSNQS WSWRSPTPHVVT 14 Phage display (common panning) 4 DOI:10.1002/adfm.200304501 Silver nanoparticles Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phages were washed in TBST buffer followed by elution with 0.2M glycine-HCl pH2.2. The phage-nanoparticles was incubated in lysis buffer to disrupt the phage coat, resulting in phage DNA release. The PCR reaction mix was directly added to the silver nanoparticles and placed in a thermocycler for PCR amplification. The PCR products were cloned into the TOPO vector. The clones were then sequenced using an automated DNA sequencer. 1453 HSVRWLLPGAHP HETNPPATIMPH WASAAWLVHSTI SPLQVLPYQGYV ESIPALAGLSDK GVLNAAQTWALS TPNSDALLTPAL HYPTLPLGSSTY HAMRPQVHPNYA QYKHHPQKAAHI YGNQTPYWYPHR HPPTDGMVPSPP TWQPFGMRPSDP TGDVSNNPNVTL 14 Phage display (common panning) 4 DOI:10.1002/adfm.200304501 Cobalt nanoparticles Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phages were washed in TBST buffer followed by elution with 0.2M glycine-HCl pH2.2. The phage-nanoparticles was incubated in lysis buffer to disrupt the phage coat, resulting in phage DNA release. The PCR reaction mix was directly added to the silver nanoparticles and placed in a thermocycler for PCR amplification. The PCR products were cloned into the TOPO vector. The clones were then sequenced using an automated DNA sequencer. 1454 HSVRWLLPGAHP SAPNLNALSAAS SVSVGMKPSPRP SPLQVLPYQGYV SLTQTVTPWAFY TNLDDSYPLHHL TPNSDALLTPAL HYPTLPLGSSTY TQQTDSRPPVLL QYKHHPQKAAHI TFPSHLATSTQP QNFLQVIRNAPR KLHSSPHTPLVQ QLLPLTPSLLQA 14 Phage display (common panning) 4 DOI:10.1002/adfm.200304501 Cobalt nanoparticles Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phages were washed several times in TBST buffer only. The phage-nanoparticles was incubated in lysis buffer to disrupt the phage coat, resulting in phage DNA release. The PCR reaction mix was directly added to the silver nanoparticles and placed in a thermocycler for PCR amplification. The PCR products were cloned into the TOPO vector. The clones were then sequenced using an automated DNA sequencer. 1455 GTSTFNSVPVRD SAPNLNALSAAS SVSVGMKPSPRP VPTNVQLQTPRS 4 Phage display (common panning) 4 DOI:10.1002/adfm.200304501 Cobalt nanoparticles Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1456 KLWTIPQ(2) KLWTIPM(1) KLWVIPQ(2) KLWSIPR(1) KVWYITP(1) KVWVLPI(1) KVFLLPR(1) KVFNWPW(1) YSLRLDY(2) FDSLVAP(1) MESQGMK(1) SQLIPWS(1) SPSWEST(1) SPTPFLL(1) KFHTHFH(1) AHSTLMR(1) ITPLAWG(1) HDIRTTH(1) K-[LV]-W-x-I-P-x 18 Phage display (common panning) 5 DOI:10.1023/A:1005605808186 Interleukin-6, IL-6 Interleukin-6 receptor 1P9M, Not determined. Ph.D.-7 phage display library (X7) A synthetic oligopeptide, KLWTIPQ, was prepared but it did not inhibit growth of MH60, an IL-6 dependent cell line. In interaction between IL-6 and its two receptors (IL-6R and gp130), a peptide with two cysteine residues at each end of consensus sequence, GGCKLWTIPQCGG (PC), was also synthesized, which significantly inhibited cell growth of MH60 at over 100 μM and phosphorylation of Stat3, which is primary signal transducer and activator of transcription, phosphorylated by IL-6 signal. 1457 VVVGSLVVARLR(2) 1 Phage display (common panning) 3 PMID:10.3923/jbs.2007.1382.1387 Banana streak virus, BSV Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phage was eluted with trypsin. 1458 SQSYPTRNS[1.006 ± 0.021] DQSYPADAN[NT] GGSQSYPDL[NT] EDGGAQSYP[NT] WRTAPESYP[NT] VQATLQSYP[NT] IHQSYPDRG[NT] REGAIQSYP[NT] RLTPESDDR[0.662 ± 0.025] QARYAKEPD[0.595 ± 0.042] QSYP, R-x(2)-P-E-P-D 10 Phage display (competitive panning) PMID:12545550 Anti-gp120 monoclonal antibody F105 Surface protein gp120, SU Not determined. Not determined. J404 phage display library (X9) ELISA Plates were coated with MAb F105 and negative control MAb F91 at 0.1 μg/well. Phage clones were added at 1.0e9 pfu/well. Parent phage M13KBst was a negative control. Phage binding was detected by rabbit anti-M13 antiserum, followed by AP-conjugated goat anti-rabbit secondary Ab. Absorbance values at 405 nm are reproduced from the bar graph and shown as the mean of duplicate samples ± standard deviations. The absorbance for the phage M13KBst was 0.053 ± 0.007. NT denotes not tested. Bound phages were eluted by the addition of rgp120. Phage display selection against MAb F105 resulted in a number of phage clones that matched the QSYP consensus sequence. Many phage clones selected by MAb F105 also matched the consensus RXXPEPD. 1459 AGPPYQSYP AMRDYQSYP SQSYPDR SQSYPD YMSYPNRSA YQSYPSREH [YS]-Q-S-Y-P, E-Q(2)-V-S-A-T-A-Q 6 Phage display (common panning) 3 PMID:12545550 Anti-rhodopsin monoclonal antibody 4B4 Rhodopsin Not determined. Not determined. J404 phage display library (X9) Phage screened against the MAb 4B4 sample resulted in the selection of two consensus sequences, with one consensus matching an epitope on the intradiskal face of rhodopsin, while the other sequence (QSYP) could not be mapped to rhodopsin. 1460 YMSYP TDWHYQSYP TDLQYQRYP TDWQYQSYP YQSYPSREN YDHNYQSYP HMSYP YQSYP, GPQV 7 Phage display (common panning) 3 PMID:12545550 Anti-cytochrome b-245 monoclonal antibody 449 Cytochrome b-245 Not determined. Not determined. J404 phage display library (X9) Phage selected during screening with the MAb 449 produced a consensus sequence GPQV matching an epitope on cytochrome b, as well as sequences containing the QSYP motif. 1461 YQSFP[NT] YMSYP[0.749 ± 0.040] YQSYV[NT] SQSYPDR[NT] TSYQSRPT[NT] GKSQYESYP[NT] GAVSYESYP[NT] NKTAYESYP[NT] YMSYPNRSA[NT] AMRDYQSYP[NT] TDWQYQSYP[0.424 ± 0.034] QKHYYESYP[NT] GDVMYMSYP[NT] YQSYP 13 Phage display (common panning) 3 PMID:12545550 Anti-MOMP monoclonal antibody GZD1E8 Major outer membrane porin, MOMP Not determined. Not determined. J404 phage display library (X9) ELISA Plates were coated with MAb GZD1E8 at 0.1 μg/well. Phage clones were added at 5e8 pfu/well. Parent phage M13KBst was a negative control. Phage binding was detected by rabbit anti-M13 antiserum, followed by AP-conjugated goat anti-rabbit secondary Ab. Absorbance values at 405 nm are reproduced from the bar graph and shown as the mean of duplicate samples ± standard deviations. The absorbance for the phage M13KBst was 0.064. NT denotes not tested. MAb GZD1E8 selected phage with the QSYP consensus, in addition to phage with a consensus sequence that maps to the C. pneumoniae major outer membrane protein. 1462 CWPLSHSVIVC(3)[1.504/1.407/1.609] CYPLNPEVYHC(2)[1.210/0.809] CSSVTAWTTGC(1)[2.304] CYMASGVFLC(1)[1.464] CWPLGPSTYIC(1)[0.865] CSLIASMETGC(1)[2.360] CYIGDPPENPC(1)[0.665] CWPLGDSTVIC(1)[1.713] CPLRLAFTFGC(1)[1.910] CTRMSHGYWIC(1)[2.015] 10 Phage display (common panning) 3 PMID:15131117 Anti-PSA monoclpnal antibody 30H12 Poly-α 2-8 sialic acid, PSA Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA Reactivity of BSA-coupled peptides with 30H12 anti-PSA Ab was measured by ELISA tests. Absorbances at 492 nm were reproduced from the bar graph and shown. BSA-coupled MK6 and MK11 reverse peptides have values of A < 0.1 (data not shown). Authors tested mimotopes bioactivity in several in vitro and in vivo tests and demonstrated their ability to enhance PSA biological activity. 1463 IQSPHFF(6) SQLSGPQ(1) MHQGSNT(1) SILPYPY(3) GTPAPVN(1) HPTHTDP(2) KLPASLT(1) YAFTPPP(1) SDMGSLR(2) QLTNLRQ(2) TASYLTL(1) TELARKI(1) TKTDTWL(5) ASLNSNS(1) 14 Phage display (subtractive panning) 3 PMID:20052979 Octyltrimethoxysilane Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) To obtain only those specific phage displayed peptides which identify the C8 ink,the eluted phages were subsequently incubated with a Si substrate, to screen out phages that bind to the background Si. 1464 LEHPFPK(1) EPLQLKM(1) GNTPSRA(3) QILAFNS(1) TFINPSH(1) GPLAKFP(1) LVQTFGK(1) HVPLLAT(1) GETRAPL(1) SILPYPY(1) YHQTTIT(1) AYSTLWP(1) AHLPPAQ(1) TTYSRFP(1) GIRHTNP(1) TTYSRFP(1) HAIYPRH(1) 17 Phage display (subtractive panning) 5 PMID:20052979 Octyltrimethoxysilane Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) To obtain only those specific phage displayed peptides which identify the C8 ink,the eluted phages were subsequently incubated with a Si substrate, to screen out phages that bind to the background Si. 1465 HAIYPRH(4) LPIWRDF(1) TTYSRFP(2) QILAFNS(2) AYSTLWP(1) SILPYPY(2) GETRAPL(1) GIRHTNP(1) VYPHPER(1) GNTPSRA(1) 10 Phage display (subtractive panning) 6 PMID:20052979 Octyltrimethoxysilane Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) To obtain only those specific phage displayed peptides which identify the C8 ink,the eluted phages were subsequently incubated with a Si substrate, to screen out phages that bind to the background Si. 1466 NSSPYLNTK(20) ASSLLASSP(16) PQSPGSSFP(10) PHNRQESPA(7) IPFPTLFAP(2) PHESDATVR(2) LVGTPNKTK(2) PSPSLSHPL(1) PPLKPVIDE(1) S(2)-[PLF] 9 Phage display (common panning) 3 PMID:7994916 Anti-TNF-α polyclonal antibody Tumor necrosis factor Not determined. Not determined. pVIII-9aa phage display library (X9) Biotinylated anti-human immunoglobulin was added to the reacted library for 3 h at room temperature and the immune complexes were immobilized in streptavidin-coated polystyrene tubes. Authors demonstrated that three synthetic peptides (ASSLLASSP, NSSPYLNTK and PPLKPVIDE) displayed by the selected phages reduced the binding of the autoantibodies to TNF-α protein by 50%. Interestingly, the sera of mice (BALB/c) immunized with phages displaying ASSLLASSP and NSSPYLNTK peptide showed an anti-TNF-α response as detected by ELISA. This response was not found in mice immunized with the wild type phage. Thus, the recombinant phages selected from the phage libraries could be used as carrier for immunization, and therefore as a tool for vaccine development. 1467 GLKVCHGPAGYPVCPCD(3) GTKRCFNVYPCQDIFVI(1) 2 Phage display (competitive panning) 2 PMID:30267550 Serum 14018 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 1468 VNWRGPSATLEGTNSNTGRRGQAVACRTCF(4) DERQIQRQEPMVRNSERDAMRCRTCAFKEL(2) TITNSASGLHFCKTCWKNSGGPAVAGKQDE(2) SLDRWPEHLATMGNRLGMTRQCKTCVGSTL(2) QRKESNPNLGCVTCGFRVRQTVGESDGGS(1) RYSGVVGNAVSEGERLNGLSSSCVTCLGWR(1) CRTCGEVGLMTRPGVRMNA(1) GAGQVERLREAKDPCRTCGGSRWRGEPFWM(1) VQWRWNDTEWMRCKTCMLSE(1) RHPHKRVRQYDGMRGAGGDWSCKTCLRPGY(1) PKRQISMERWLQVTQGEEVTPCATCNPWVA(1) LCKTCVRSHQERTVKGDQVTGTRICQTC(1) WDKRPVVWLRFEESQRLSRCATCGVGGVE(1) NVNEPGIRQGPAASVGWKVVRLAGICKTCV(1) GMKIVVFPKRSVPDVTGSQGAPPCRTCTST(1) SVLRQAAQFGNFELYVRREGNCRALTGCMR(1) C-[RK]-T-C 16 Phage display (common panning) 2 PMID:8700874 Anti-HBsAg monoclonal antibody H166 Hepatitis B surface antigen Not determined. Not determined. fNG1 phage display library (X30) 1469 EQFAKTCPMQAVKGGWASTLCRSVYSGNVR(2) RVAKGQGPLRVYLTQRRKQGNASWEWEEFI(2) LTEVVISRSADCRFRDVITGECCGWHRGCF(2) 3 Phage display (common panning) 2 PMID:8700874 Anti-HBsAg monoclonal antibody H5 Hepatitis B surface antigen Not determined. Not determined. fNG1 phage display library (X30) 1470 SPVSYGEWRARYCTNGGQGTVQRRADRSCW(23) RASGARARCEHRSGLSLSWQPSECSDSRTT(1) 2 Phage display (common panning) 2 PMID:8700874 Anti-HBsAg monoclonal antibody H35 Hepatitis B surface antigen Not determined. Not determined. fNG1 phage display library (X30) 1471 RGGLMRGGSSDTRLMGWQSSSIYSFQARGS(10) GSVPLSRRKEVWAGEEESFGYWLVNWQEMM(5) GRKTEKYSDGWTSMHSAEVCTQWNMSYCMI(3) GVELGKRANRGGSTTSWHGSSLGDIQSYWT(2) 4 Phage display (common panning) 2 PMID:8700874 Anti-HBsAg monoclonal antibody H53 Hepatitis B surface antigen Not determined. Not determined. fNG1 phage display library (X30) 1472 SLIPRGARTPRQCRSACPPREYSSADHWKS AGMWRVPRAENYSAPVRTRPKRQPWAQGSY LSADTLRSNSVDHDRVQNEVRSSRERRQPR LLEVGRDWVGGNNMWVGRMRERRKNERRQF IWDRRRQRQPGRVENYPVGKPASHQLYILN MKLPVDENKSGRRERRQPTPAGERELIRFD YLPYRAGVDKSGGREVGSHMRFFTRERRHA AELTGYEDVRRVEKRKGLAGTRERRQPAAY KMLRERRQPSSNLDYEKEVGQFYVVVAKSD SYGSGAARVSKLQETGGRRGRRQPSGSYIG FPARSEASGRQRRQPGRDVTHGEAVRVNIL IGPRRKWDALASDSGCNPSSHSQRCHRLKP PCQNTYRGLMLNEDCRQGRRTRRQPPATTL IGQGQQKEALGSRQRFDLRGRRQPVGSGKW RSGGVSELRAEGVNRRQARRKEQRRQPPRY RGSYDRRRERRQPRGLR QAVVSGERGARPRRQPRTPGVAACARSAGG QGYVDAGSISRFGGRGVWRQRRQPLLNGSF KIVRLTDAGHRTRRQPRTEEMWKVSTWLLN QVLKGGVSKRTMKGRACRQQACPKTVPSVV KELHTVEANERLKLREGRLRDRRQPISQWN R-G-R(2)-Q-P 21 Phage display (common panning) 4 PMID:8543161 Anti-HCV core protein monoclonal antibody 14-153-462 Hepatitis C core protein Not determined. Not determined. fNG1 phage display library (X30) 1473 PAPRSQESVGQTRVRVGRCEASRLGRSWCQ[1.621] GKVAARKSCTKFGRSCKVQTLWEGAGIPFI[0.600] ASARAYGECKYGRCTFNGIALPHSTMVKNP[1.516] PHWEGKVGCKRTRQGRASCSRVELDTKWGG[0.537] LARGGTSSMGWIEQGEVRWGCHNGRCSRIG[0.763] x-G-R 5 Phage display (common panning) 4 PMID:8543161 Anti-V3 loop of HIV gp120 monoclonal antibody 1001 V3 loop of HIV gp120 Not determined. Not determined. fNG1 phage display library (X30) ELISA Affinities of gpl20-mimic phages to anti-HIV mAb 1001 were measured by ELISA. Absorbance at 490 nm was determined, reproduced from the graph and shown. Phage fUSE2 was used as the negative control (with A490 of 0.052). The positive control well was coated with 0.5 μg of gpl20 (with A490 of 0.801). The results show that phage obtained are able to capture the anti-HIV mAb 1001. 1474 YPYPVFVH(1) YPYYQYFM(1) YPYGSYWA(1) VWYPYGAG(1) VWYKYPGW(1) DVFYPYPY(4) NRVWYPYG(1) NCQGTACS(1) LRCGWGVC(1) YNMYSTAA(1) AVNGCRHD(1) SYTHVASS(1) KGQAELLR(1) VFTDQKAQ(1) LNDNSAGY(1) WARNTSHS(1) YPY 16 Phage display (competitive panning) 3 PMID:1608948 Concanavalin-A, Con A Methyl α-D-mannopyranoside Not determined. Not determined. X8 fAFF1 phage display library Adherent phage were eluted with buffer containing methyl α-D-mannopyranoside or yeast mannan at concentrations sufficient to block rebinding of peptide at the sugar binding site. 1475 YNYGWVEF(5) YRYDIFRE(1) YDYGSFSK(1) YSYPYYHL(1) NYDYMGIW(1) YPYAYIWT(1) VVFYDYGS(1) YEAHYQYG(1) GTWFTNFR(1) SRCGLLVE(1) Y-x-Y 10 Phage display (common panning) 3 PMID:1608948 Concanavalin-A, Con A Methyl α-D-mannopyranoside Not determined. Not determined. X8 fAFF1 phage display library Biotinylated Con A was bound in 96-well microtiter plates coated with streptavidin. Each selection was carried on the plates. Bound phages were eluted with a low-pH citrate buffer (pH 3.0) to denature the receptor-ligand interaction. 1476 HWSPPSL(1) EQTINQW(1) LRPDSIF(1) HFTTRLL(1) YLTTDEF(1) RLVTDEI(1) LETDELH(1) AIWQHGV(1) QVARTSF(1) WHQVSLT(1) THQRPAT(1) EHQTGTW(1) KSFNSPH(1) SGTSHIR(1) AIWYPHV(1) TPHNPAI(1) EHPWPLL(1) 17 Phage display (common panning) 2 PMID:9880034 Anti-CMV CP polyclonal antibody Capsid protein, CP Not determined. Not determined. Ph.D.-7 phage display library (X7) From the second round of biopanning, 12 of 17 clones (70%) were found to contain insert sequences matching with at least three amino acids in the primary sequence of the CMV CP. 1477 QLVTDEL(3)[0.18] EHQLNRA(2)[NT] YHQTTIP(2)[NT] SLTTDEL(1)[0.20] GHQVSRL(1)[NT] HGLHLPV(1)[NT] VLPDSVW(1)[NT] SLPTLTL(1)[NT] KIPIALS(1)[NT] HAIYRPH(1)[NT] FHQMPDA(1)[NT] ADCTTPT(1)[NT] FGGDADR(1)[NT] SGIAVNP(1)[NT] HSYTLMF(1)[NT] GHWTRFA(1)[NT] 16 Phage display (common panning) 3 PMID:9880034 Anti-CMV CP polyclonal antibody Capsid protein, CP Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding of the positive clones to Fny-CMV polyclonal antibodies was confirmed by ELISA. Absorbance at 405 nm was determined and average ELISA value was shown. The average ELISA value of the positive clones was at least three times as high as that of a negative control clone (with average A405 of 0.06). These results suggest that the sequence LXTDEL, corresponding to amino acid residues 194-199, is a dominant epitope in the Fny-CMV CP. NT denotes not tested. Nine of 19 clones (47%) from the third round of biopanning carried insert sequences resembling a sequence of the CMV CP. 1478 SPSAPTH(4) SSFSPST(3) ALSIIGK(1) SASIRPQ(1) SLGMHPL(1) 5 Phage display (common panning) 3 PMID:9880034 Anti-PSV-J polyclonal antibody J strain of peanut stunt virus Not determined. Not determined. Ph.D.-7 phage display library (X7) 1479 SHLSTMV(4) SHLSTML(4) AHKSTLR(2) AHRPTLL(2) LERTPGK(1) LPYTMWV(1) TASFHRN(1) AHIPWLL(1) 8 Phage display (common panning) 3 PMID:9880034 Anti-TAV-V polyclonal antibody V strain of tomato aspermy virus Not determined. Not determined. Ph.D.-7 phage display library (X7) 1480 LTMTSPI(5) AQGNSVK(2) APWELPL(1) 3 Phage display (common panning) 3 PMID:9880034 Anti-TAV-C polyclonal antibody C strain of tomato aspermy virus Not determined. Not determined. Ph.D.-7 phage display library (X7) 1481 KQTWQQLWD(21) VPHPTWWRG(3) VCQAWPCKL(2) WTRSDHRIQ(2) KQTQEQLWD(1) SQTWRTAFL(1) KDWWSTGEV(1) GGHTWKMLW(1) NQTWQQLWD(1) YKTSWDT(1) KQGSAWWHR(1) YPAWHSSAW(1) AKITSQRMM(1) KMYQQFVLD(1) QNGPTWWRW(1) YNSPTWWRA(1) MYWPTWYRG(1) NGSPTWWRS(1) EGADLEFRR(1) LDLPAWHGR(1) HAKTPPWWR(1) SNIPSWYRW(1) VESRPWWWR(1) AHSPVWWRQ(1) VESPSWWRR(1) QDKVPLWWR(1) GDQWRAWLT(1) WQNWRSAFH(1) SEEVWSWRS(1) NLWYREEWR(1) GGGQDWRR(1) EEKIWRTQA(1) WTRDQHQIH(1) WTLREHDFH(1) WQITQHKLQ(1) WTLQHHRVV(1) WTLGEHTLI(1) WRLSDHRMV(1) WTIKDHQLL(1) WKLSEHRMA(1) WSLGQHRIF(1) HGKHTHKVG(1) HGDKHKHRG(1) KPHQHKVHK(1) 44 Phage display (common panning) 3 PMID:10211822 Anti-F-actin polyclonal antibody Actin, alpha skeletal muscle Not determined. Not determined. J404 phage display library (X9) Western blot,Competition experiment The result of Western blot demonstrated the specificity of the recognition of KQTWQQLWD by the anti-actin antiserum. Besides, the KQTWQQLWD-ovalbumin conjugate inhibited antibody binding to actin by 58 ± 7% (n=4). The scrambled peptide conjugate, WQDKWLQTQ-ovalbumin, inhibited binding only by 12 ± 2% (n=5). One hundred and forty-four positively staining clones identified in this way were sequenced. Of these, 54 displayed peptides with sequence similarities. When the most abundantly selected sequence, KQTWQQLWD, was produced as a synthetic peptide and derivatized to ovalbumin, the complex was strongly recognized by the antiserum on Western blots and inhibited the binding of the antibody to immobilized F-actin by 60%. A scrambled version of this sequence WQDKWLQTQ, when coupled to ovalbumin, was not recognized by the antiserum and minimally inhibited binding of antiserum to immobilized F-actin by 10%. KQTWQQLWD contained four residues that corresponded, in frame, to a highly conserved six residue region of the chicken β-actin sequence (351)TFQQMW(356). 1482 ETQRCTWHMGELVWCEREHN KEASCSYWLGELVWCVAGVE GELVW 2 Phage display (common panning) PMID:10678837 Ig gamma-1 chain C region Not determined. Not determined. Not determined. X(i)CX(j)CX(k) M13 phage display library pool The gene sequence for KEASCSYWLGELVWCVAGVE was transferred to gene III of M13 bacteriophage and improved by monovalent phage display. Five residue blocks were randomly mutated in six separate libraries to exhaustively cover the noncysteine positions in the peptide sequence. Preferred residues from selection of these libraries for binding to Fc were then recombined to give three more libraries spanning the peptide sequence (13). Selection patterns from these libraries suggested a 13-residue core Fc binding sequence (DCAWHLGELVWCT). Crystal structure of DCAWHLGELVWCT, in complex with IgG-Fc, has been deposited in the PDB under accession number 1DN2. 1483 RRTPDPAVSPWQLTY RDGQRLTSSKTMLPY KLTSSLRYNSPPLCF KSTSSLRDNSPPVCF KLTSSLRCNCPPLRF THDLSSRASSSLSYN QAPRLMSSLSYFPQS DHRSPPWLTSLLTIS DTWPTARLTSSMQYI HTYTSHLRYVPPISL GHRYTSSVSLTEACP SGTSHSASTTSKWFL NLLSVAPFWPLNDSL SDPDQWPFWRANEYG R-L-T-S(2)-L-R-Y-N-P 14 Phage display (common panning) 3 PMID:11157855 Anti-dsDNA polyclonal antibody Deoxyribonucleic acid, DNA Not determined. Not determined. f88-15mer phage display library (X15) Three rounds of screening were performed with the biotinylated anti-dsDNA antibody. For the first round of selection, 10μg of biotinylated anti-dsDNA antibodies in 400 μl of 0.5×TBST (25 mM Tris, 75 mM NaCl and 0.05% Tween 20, pH 7.5) containing 1 mg/ml dialyzed BSA were immobilized onto a streptavidin-coated Petri dish. In the subsequent rounds of selection, 1μg of biotinylayed anti-dsNDA antibodies were used. Synthesized peptide RLTSSLRYNP could be recognized by 88% (37 out of 42) of anti-dsDNA antibody-positive SLE sera, suggesting that the mimotope is shared by ds and ssDNAs as well as native RNA, whereas denatured RNA was not observed to inhibit the binding. The peptide was also to elicit an immune response in rabbits and the anti-peptide rabbit serum was observed to cross-react with the peptide, ss and dsDNAs, and ss and dsDNAs could inhibit the binding of the anti-peptide serum and the peptide. However, the inhibition was not obtained with RNA. 1484 RQHPKL(35) 3 Phage display (common panning) 3 PMID:7576084 Anti-lymphotoxin polyclonal antibody Lymphotoxin Not determined. Not determined. f3-6mer phage display library (X6) Phages displaying a ligand for the biotinylated polyclonal antibodies are captured on the dish by binding of the biotin moiety to immobilized streptavidin. 1485 LPSRQHPRSNAYRPF(5) PRQHPTLDRAVRLGS(4) GWRGVHQFGSPRQHR(5) PRQLHGLHWNFSPVR(1) RGWMGAAAMLLDPPN(2) IYNPLSLLKWELPFP(l) KHAFADSTAKPWVGR(1) 7 Phage display (common panning) 3 PMID:7576084 Anti-lymphotoxin polyclonal antibody Lymphotoxin Not determined. Not determined. X15 phage display library Phages displaying a ligand for the biotinylated polyclonal antibodies are captured on the dish by binding of the biotin moiety to immobilized streptavidin. 1486 HAIYPRH(6) 1 Phage display (subtractive panning) 10 PMID:11277922 Transferrin receptor protein 1 Iron-loaded human transferrin Not determined. Not determined. Ph.D.-7 phage display library (X7) In fact, the target is CEF+hTfR cells expressing the human transferrin receptor. The original phage library was applied to CEF cells. Unbound phage were transferred to another well of CEF cells, before transferring the unbound phage to a well of CEF+hTfR cells. In the final round of biopanning selection, the HAIYPRH phage represented 67% (6/9) of the phages sequenced. Phage containing the sequences HAIYPRH were inhibited from binding the hTfR in a dose-dependent fashion when peptides of the same sequence were present in a competition assay. Interestingly, transferrin did not compete with either of these sequences for receptor binding, suggesting that these peptides bind a site on the hTfR distinct from the transferrin binding site. 1487 THRPPMWSPVWP(8) 1 Phage display (subtractive panning) 7 PMID:11277922 Transferrin receptor protein 1 Iron-loaded human transferrin Not determined. Not determined. Ph.D.-12 phage display library (X12) In fact, the target is CEF+hTfR cells expressing the human transferrin receptor. The original phage library was applied to CEF cells. Unbound phage were transferred to another well of CEF cells, before transferring the unbound phage to a well of CEF+hTfR cells. In the final round of biopanning selection, the THRPPMWSPVWP phage represented 89% (8/9) of the phages isolated and sequenced. Phage containing the sequences THRPPMWSPVWP were inhibited from binding the hTfR in a dose-dependent fashion when peptides of the same sequence were present in a competition assay. Interestingly, transferrin did not compete with either of these sequences for receptor binding, suggesting that these peptides bind a site on the hTfR distinct from the transferrin binding site. 1488 FHENWPS(5)[15.708 ± 7.915] GPLYHTP(2)[26.697 ± 9.371] WSLDPHR(1)[5.454 ± 1.617] 3 Phage display (common panning) 3 PMID:11479280 Anti-β-1,2-linked mannoside monoclonal antibody DJ2.8 β-1,2-linked mannoside Not determined. Not determined. Ph.D.-7 phage display library (X7) Densitometry The intensity of bound mAb was measured and analysed by densitometry. Data shown were intensity of bound mAb (arbitrary units) and reproduced from FIG. 3 in the reference. Results are expressed as the mean ± SD of measurements from one representative experiment. It revealed that all clones were recognized by mAb although different levels of binding were observed. Sixty percent of the phages had an identical DNA insert corresponding to the peptide sequence FHENWPS that was recognized specifically by the mAb. Injection of KLH-coupled peptide into mice generated high titers of polyclonal antibodies against C.albicans yeast cell walls. The anti-FHENWPS antibodies bound to C.albicans PPM and were inhibited by soluble β-1,2-mannotetraose. 1489 SEVGCRAGPLQWLCEKYF(6) 1 Phage display (common panning) 4 PMID:9636028 Insulin-like growth factor-binding protein 1, IGFBP-1 Insulin-like growth factor I, IGF-I 2DSQ, Not determined. SGTACX(2)GPX(4)CSLAGSP M13 phage display library ELISA Peptides CRAGPLQWLCEKYFG (p1-01, cyclic) and SEVGCRAGPLQWLCEKYFG (p1-02, cyclic), both based upon the predominant phage clone (SEVGCRAGPLQWLCEKYF), showed the highest affinities for IGFBP-1, blocking IGFBP-1 binding to IGF-1 with IC50s of 180 and 50 nM, respectively. The initial selection was carried out by binding phage, washing, and then eluting by incubation with 50 mM DTT (to reduce the biotin-disulfide linkage, releasing phagemid particles) for 1 h at room temperature. In the second and third cycles of binding selection, streptavidin was included in the phage cocktails along with biotin, and 5 g/L ovalbumin, or 5 g/L instant milk in 50 mM sodium carbonate buffer, was used as the blocking agent. The fourth round was carried out on plates directly coated with IGFBP-1 or with albumin only. 1490 SEVGCRAGPLQWLCEKYF(5) KDPVCGEGPLMRICERLF(1) 2 Phage display (common panning) 4 PMID:9636028 Insulin-like growth factor-binding protein 1, IGFBP-1 Insulin-like growth factor I, IGF-I 2DSQ, Not determined. X(4)CX(2)GPX(4)CX(4) M13 phage display library ELISA Peptides CRAGPLQWLCEKYFG (p1-01, cyclic) and SEVGCRAGPLQWLCEKYFG (p1-02, cyclic), both based upon the predominant phage clone (SEVGCRAGPLQWLCEKYF), showed the highest affinities for IGFBP-1, blocking IGFBP-1 binding to IGF-1 with IC50s of 180 and 50 nM, respectively. Peptide p1-13, corresponding to phage clone Ф13 (KDPVCGEGPLMRICERLF), bound IGFBP-1 much more weakly, blocking IGF-1 binding with IC50 of 5.4 μM. The initial selection was carried out by binding phage, washing, and then eluting by incubation with 50 mM DTT (to reduce the biotin-disulfide linkage, releasing phagemid particles) for 1 h at room temperature. In the second and third cycles of binding selection, streptavidin was included in the phage cocktails along with biotin, and 5 g/L ovalbumin, or 5 g/L instant milk in 50 mM sodium carbonate buffer, was used as the blocking agent. The fourth round was carried out on plates directly coated with IGFBP-1 or with albumin only. 1491 EVDGRWWIVETFLAKWDHMA(6) 1 Phage display (common panning) 4 PMID:9636028 Insulin-like growth factor-binding protein 1, IGFBP-1 Insulin-like growth factor I, IGF-I 2DSQ, Not determined. X20 M13 phage display library ELISA Peptide p1-31, corresponding to phage clone Ф31 (EVDGRWWIVETFLAKWDHMA), bound IGFBP-1 much more weakly, blocking IGF-1 binding with IC50 of > 10 μM (data not shown). The initial selection was carried out by binding phage, washing, and then eluting by incubation with 50 mM DTT (to reduce the biotin-disulfide linkage, releasing phagemid particles) for 1 h at room temperature. In the second and third cycles of binding selection, streptavidin was included in the phage cocktails along with biotin, and 5 g/L ovalbumin, or 5 g/L instant milk in 50 mM sodium carbonate buffer, was used as the blocking agent. The fourth round was carried out on plates directly coated with IGFBP-1 or with albumin only. 1492 EVRSFCTDWPAEKSCKPLRG[+++] 1 Phage display (common panning) 4 PMID:12119302 Human serum albumin Not determined. Not determined. Not determined. CX(2)GPX(4)C, X(4)CX(2)GPX(4)CX(4), and X(i)CX(j)CX(k) M13 phage display library pool ELISA In phage ELISA, the absorbance at 405 nm was monitored. 1493 RAPESFVCYWETICFERSEQ[++] 1 Phage display (common panning) 4 PMID:12119302 Human serum albumin Not determined. Not determined. Not determined. X20 and X(i)CX(j)CX(k) M13 phage display library pool ELISA In phage ELISA, the absorbance at 405 nm was monitored. 1494 EMCYFPGICWM[+++] 1 Phage display (common panning) 4 PMID:12119302 Human serum albumin Not determined. Not determined. Not determined. X8 and X(2)CX(j)CX(2) M13 phage display library pool ELISA In phage ELISA, the absorbance at 405 nm was monitored. 1495 GENWCDSTLMAYDLCGQVNM[+++] 1 Phage display (common panning) 4 PMID:12119302 Rabbit serum albumin Not determined. Not determined. Not determined. CX(2)GPX(4)C, X(4)CX(2)GPX(4)CX(4), and X(i)CX(j)CX(k) M13 phage display library pool ELISA In phage ELISA, the absorbance at 405 nm was monitored. 1496 MDELAFYCGIWECLMHQEQK[+++] 1 Phage display (common panning) 4 PMID:12119302 Rabbit serum albumin Not determined. Not determined. Not determined. X20 and X(i)CX(j)CX(k) M13 phage display library pool ELISA In phage ELISA, the absorbance at 405 nm was monitored. 1497 DLCDVDFCWF[+++] 1 Phage display (common panning) 4 PMID:12119302 Rabbit serum albumin Not determined. Not determined. Not determined. X8 and X(2)CX(j)CX(2) M13 phage display library pool ELISA In phage ELISA, the absorbance at 405 nm was monitored. 1498 KSCSELHWLLVEECLF[+++] 1 Phage display (common panning) 4 PMID:12119302 Rabbit serum albumin Not determined. Not determined. Not determined. X(2)CX(j)CX(2) M13 phage display library ELISA In phage ELISA, the absorbance at 405 nm was monitored. 1499 RNEDPCVVLLEMGLECWEGV[+++] 1 Phage display (common panning) 4 PMID:12119302 Rat serum albumin Not determined. Not determined. Not determined. CX(2)GPX(4)C, X(4)CX(2)GPX(4)CX(4), and X(i)CX(j)CX(k) M13 phage display library pool ELISA In phage ELISA, the absorbance at 405 nm was monitored. 1500 QRQMVDFCLPQWGCLWGDGF[+++] 1 Phage display (common panning) 4 PMID:12119302 Rat serum albumin Not determined. Not determined. Not determined. X20 and X(i)CX(j)CX(k) M13 phage display library pool ELISA In phage ELISA, the absorbance at 405 nm was monitored. 1501 DLCLRDWGCLW[+++] 1 Phage display (common panning) 4 PMID:12119302 Rat serum albumin Not determined. Not determined. Not determined. X8 and X(2)CX(j)CX(2) M13 phage display library pool ELISA In phage ELISA, the absorbance at 405 nm was monitored. 1502 DTCVDLVRLGLECWG[+++] 1 Phage display (common panning) 4 PMID:12119302 Rat serum albumin Not determined. Not determined. Not determined. X(2)CX(j)CX(2) M13 phage display library ELISA In phage ELISA, the absorbance at 405 nm was monitored. 1503 MSRNFSGTTPRHLLL(2) SVHQLNAPRSLLFNH(1) LPPRNLLYGNSRLDW(1) YRPISNMPRQLLMRL(1) MDTPRHLMAYLITQR(2) LMNSPRQLMTRKMSH(1) TIPSAKSPRSLFVTH(1) STSPPRSLLGSLDRT(1) LTHHPHYPRHLMGSR(1) LPPSEASTPRRLLHQ(1) PRSLLSPSDSLRFIT(1) PRNLFLPTAHPFST(1) TEFITREARHLLLIP(1) FQEPKSEQRHLFYKW(2) P-R-H-L(2) 14 Phage display (common panning) 3 PMID:7507752 Anti-ATPase alpha-1 subunit monoclonal antibody M8-P1-A3 Sodium/potassium-transporting ATPase subunit alpha-1 Not determined. Not determined. X15 M13 phage display library The M8-Pl-A3 epitope was found to consist of the five amino acid 494-PRHLL-498 sequence stretch of α, with residues PRxLx being critical for antibody recognition. 1504 KFYRFL(7) DRFRHI(2) KFDRFL(1) KLYRFL(1) KLYGFV(1) DRFRQV(1) EFDRFR(1) QKPIKL(1) DLLRGA(1) NMPKPI(1) KFDRFL, DRFR 10 Phage display (common panning) 3 PMID:8956081 Anti-sperm whale myoglobin polyclonal antibody Myoglobin Not determined. Not determined. f3-6mer phage display library (X6) Phages displaying a ligand for the biotinylated polyclonal antibodies are captured on the dish by binding of the biotin moiety to immobilized streptavidin. Motif KFDRFL, which matches amino acid residues 42-47 the sperm-whale myoglobin sequence. 1505 RFDRLK(8) LELFRR(4) RFDWLK(1) GFDRLK(1) LELFRG(1) LGLFRR(1) RCDRLM(1) RVDRLM(1) LEVCSE(1) GCDLYI(1) RFDRLK, LELFRR, DRLM 10 Phage display (common panning) 3 PMID:8956081 Anti-sperm whale myoglobin polyclonal antibody Myoglobin Not determined. Not determined. f3-6mer phage display library (X6) Phages displaying a ligand for the biotinylated polyclonal antibodies are captured on the dish by binding of the biotin moiety to immobilized streptavidin. Two motifs show homology to the sperm-whale myoglobin sequence: RFDRLK, which matches amino acid residues 45-50 (RFKHLK), and LELFRR, which matches amino acid residues 135-140 (LELFRK). 1506 YWYRWS(5) HWYRFM(3) QRYRFH(2) WLYRFS(2) QLYRFH(1) YFYRFR(1) KFYRFL(1) QWYRFM(1) FWNRFA(1) KWNRFA(1) HWNRFV(1) VWWRFM(1) YLYRPY(1) HWYRFM, QRYRFH, YWYRWS,YRF, WNRF 13 Phage display (common panning) 3 PMID:8956081 Anti-sperm whale myoglobin polyclonal antibody Myoglobin Not determined. Not determined. f3-6mer phage display library (X6) Phages displaying a ligand for the biotinylated polyclonal antibodies are captured on the dish by binding of the biotin moiety to immobilized streptavidin. 1507 REDLFDRYAHLRTPV QTNVKYDRFDWLLRQ KYDRFVIYKDGSVQT ELREEYKQLGYTGPH 4 Phage display (common panning) 2 PMID:8956081 Anti-sperm whale myoglobin polyclonal antibody Myoglobin Not determined. Not determined. X15 phage display library Phages displaying a ligand for the biotinylated polyclonal antibodies are captured on the dish by binding of the biotin moiety to immobilized streptavidin. Twenty-six samples were sequenced after two rounds of screening. It was found that except for 19% (5/26) of the positive clones containing the insert sequence of D-R-F, there were a few dominant sequence motifs. However, with the help of the Blitz electronic mail server, four sequences were found to show homologies to the myoglobin sequence. Three of the insert peptide sequences (clones REDLFDRYAHLRTPV, QTNVKYDRFDWLLRQ and KYDRFVIYKDGSVQT) showed similarities to amino acid residues 42-50 of the myoglobin sequence, On the other hand, the peptide insert sequence of clone ELREEYKQLGYTGPH was similar to amino acid residues 141-153 of the myoglobin sequence (which was located in the C-terminus of the protein). 1508 RLAPEPDDPITPMTK(2)[1.25 ± 0.06, 22 ± 3%] KLLPEDDESRTYHTV(1)[1.32, 19%] TQSYPPPPAWRAASR(1)[2.11, 53%] QLSPESDYDDHGMRY(1)[3.00, 40%] KLFPEEDEMRTETQR(1)[1.60, 24%] SQSYPEPARGSVPMP(1)[3.00, 29%] L-x-P-E-x-D, QSYP, E-x-[ED]-P(2)-V 6 Phage display (common panning) 3 PMID:12501244 Anti-TPO monoclomal antibody T13 Thyroid peroxidase, TPO Not determined. Not determined. X15 phage display library ELISA Absorbance at 490 nm corresponding to the binding of each clone to aAb T13 was shown. The binding of each clone to aAb T13 was inhibited by soluble human TPO at 90 nM; only clones showing an inhibition greater than 15% with hTPO are indicated. When a clone was represented at least twice, the absorbance and percent inhibition values are given as the mean value ± S.D. Mimotope sequence alignment on the TPO molecule, together with the binding analysis of the T13 aAb on TPO mutants expressed by Chinese hamster ovary cells, demonstrated that regions 353-363, 377-386, and 713-720 from the myeloperoxidase-like domain and region 766-775 from the complement control protein-like domain are a part of the IDR recognized by the recombinant aAb T13. Furthermore, authors demonstrated that these regions were involved in the binding to TPO of sera containing TPO-specific autoantibodies from patients suffering from Hashimoto\'s and Graves\' autoimmune diseases. 1509 GQSYPPRPDTSLHVT(1)[0.50, 28%] ELNPEPDTEVFPMTF(1)[0.91, 38%] KNSRQSYPEPAPVYH(1)[0.59, 28%] ENEPPVWTTESKLQS(1)[0.74, 31%] ESDPPVASYQWRLIN(1)[0.79, 36%] ESVMQSYPPHLQIPG(1)[0.50, 24%] L-x-P-E-x-D, QSYP, E-x-[ED]-P(2)-V 6 Phage display (competitive panning) 4 PMID:12501244 Anti-TPO monoclomal antibody T13 Thyroid peroxidase, TPO Not determined. Not determined. X15 phage display library ELISA Absorbance at 490 nm corresponding to the binding of each clone to aAb T13 was shown. The binding of each clone to aAb T13 was inhibited by soluble human TPO at 90 nM; only clones showing an inhibition greater than 15% with hTPO are indicated. When a clone was represented at least twice, the absorbance and percent inhibition values are given as the mean value ± S.D. Bound phages were eluted by competition with a TPO solution. Mimotope sequence alignment on the TPO molecule, together with the binding analysis of the T13 aAb on TPO mutants expressed by Chinese hamster ovary cells, demonstrated that regions 353-363, 377-386, and 713-720 from the myeloperoxidase-like domain and region 766-775 from the complement control protein-like domain are a part of the IDR recognized by the recombinant aAb T13. Furthermore, authors demonstrated that these regions were involved in the binding to TPO of sera containing TPO-specific autoantibodies from patients suffering from Hashimoto\'s and Graves\' autoimmune diseases. 1510 FSLSKPP(21)[0.468 ± 0.011] STQAMFQ(9)[0.523 ± 0.047] HSMQLST(4)[0.436 ± 0.047] HAIYPRH(3)[0.782 ± 0.022] 4 Phage display (common panning) 3 PMID:12951773 A549 bronchial epithelial cell line Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA In phage ELISA, absorbance at 420 nm was measured. Data shown were reproduced from the bar graph and expressed as means ± standard deviations. The absorbance of the negative control phage was 0.379 ± 0.024. Results demonstrated the isolated phage displaying HAIYPRH, STQAMFQ, and FSLSKPP show significantly increased levels of binding to A549 cells compared with a control wild-type phage displaying no peptide on its surface coat. 1511 NVESQPGGQPNT QYTDHHSSLLGP LYRPSDSSLAGP MNVTLSSSLDGP NGNRSPTTLMGP DRLLTTLSGPAQ Y-x(4)-S(2)-x(2)-G-P, S(2)-x(2)-G-P, T(2)-x(2)-G-P 6 Phage display (common panning) 3 PMID:14629623 Anti-p34 polyclonal antibody 34 kDa antigenic protein Not determined. Not determined. Ph.D.-12 phage display library (X12) Immunization of mice with these peptides elicited an anti-p34 antibody response. Two B-cell epitopes were identified and characterized. Based on the reactivity and the type of immune response elicited, epitope A was determined to be conformational, whereas epitope B was demonstrated to be sequential. Both epitopes were shown to be present in p34 proteins from M. avium ssp. avium or M. paratuberculosis but absent from M. intracellulare, the other member of the M. avium complex. Furthermore, both epitopes were mapped to regions of p34 that display high variability when compared to homologous proteins from other mycobacterial species of public and animal health importance. 1512 SHSALTVPLYHA WSDMSMYSHLMP MSDSHYNSQSSV SH 3 Phage display (common panning) 3 PMID:11932389 Anti-PRRSV neutralizing monoclonal antibody ISU25-C1 Sequential epitope on GP5 Not determined. Not determined. Ph.D.-12 phage display library (X12) The resin was washed with TBS containing 0.05% Tween 20 in the first panning, TBS containing 0.1% Tween 20 in the second panning, and TBS containing 0.5% Tween 20 in the third panning. Phages bound to ISU25-C1 were eluted with 100 mM glycine, pH 2.5. 1513 SHITSYHPAYFW[0.500 ± 0.003] 1 Phage display (common panning) 3 PMID:11932389 Anti-PRRSV neutralizing monoclonal antibody ISU25-C1 Sequential epitope on GP5 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Results of phage ELISA are shown above and expressed as corrected OD at 405 nm (OD for selected phage - OD for unselected phage library) ± standard deviation. In the second panning, a CNBr-activated Sepharose-coupled MAb was used. In the third panning, protein G was used again to capture the phage-antibody complexes from solution. The resin was thoroughly washed with 10 mM Tris, pH 7.5, in the first panning, with 10 mM Tris, pH 7.5, containing 0.5 M NaCl in the second panning, and with 10 mM Tris, pH 7.5, containing 0.05% Tween 20 in the third panning. 1514 ALVNIPISNNLA[0.045 ± 0.008] ALVNWHGVYNYR[NT] ALVNNSHTMPLW[NT] ALVNSPLTRAPM[NT] GGYPQQMVRHFA[NT] KIPYNLSFMVPP[NT] ALVN 6 Phage display (common panning) 3 PMID:11932389 Anti-PRRSV polycolonal antibody Prcine reproductive and respiratory syndrome virus, RSV Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Results of phage ELISA are shown and expressed as corrected OD at 405 nm (OD for selected phage - OD for unselected phage library) ± standard deviation. NT denotes not tested. Anti-PRRSV affinity-purified antibodies with high SN titer were used to select phages from the library, and several clones carrying amino-terminal motif ALVN were recovered. This is a consensus motif found in residues 27 to 30 of PRRSV GP5. Another clone (KIPYNLSFMVPP) selected with the swine antibodies with high SN titer expressed a peptide with a YNL motif, which is also a consensus motif found in amino acids 43 to 45 of PRRSV GP5. Finally, clone KIPYNLSFMVPP carried an HF motif, which is found in residues 38 to 39 of GP5 of a PRRSV IA 97-7895 isolate, which was the primary PRRSV strain used to immunize the pigs. 1515 YKNTHLDLIYNA[0.080 ± 0.001] NSAVHFQRQFSQ[NT] 2 Phage display (common panning) 3 PMID:11932389 Anti-peptide SGSGANNSSSSHFQSIYNC polyclonal antibody Prcine reproductive and respiratory syndrome virus, RSV Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Results of phage ELISA are shown and expressed as corrected OD at 405 nm (OD for selected phage - OD for unselected phage library) ± standard deviation. NT denotes not tested. Anti-peptide S2 affinity-purified antibodies were used to select phages from the library, and 4 out of 12 selected clones corresponded to a single clone (YKNTHLDLIYNA), containing an HLXLIYN motif. This motif is also found between amino acids 38 and 44 of the consensus sequence of PRRSV GP5. Another clone selected with these antibodies (NSAVHFQRQFSQ) contained an HFQ motif located between residues 38 and 40 of GP5 of the PRRSV Iowa strain. 1516 APWHLSSQYSRT(11)[0.932 ± 0.046] HGEVPRFHAVHL(6)[0.952 ± 0.089] HPVTRFHNPVEY(2)[0.744 ± 0.026] LISSATPFSPNK(1)[0.606 ± 0.089] WSSGMTPDTGAP(1)[0.661 ± 0.020] V-[PT]-R-F-H-[AP]-V, [MA]-T-P 5 Phage display (subtractive panning) 4 PMID:20508256 Rhesus monkey neural stem cells (R-NS cells) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA In phage ELISA, the absorbance at 492 nm was determined. Data were reproduced from the graph and shown as means ± standard derivations. The absorbance of M13 wild-type phage (Wt) as negative control was 0.019. Besides, results of a competitve assay showed that the synthetic HGE peptide (HGEVPRFHAVHL) could compete with the HGE phage, achieving 50% inhibition at the concentration of about 500 nM. Further results of Western blot showed that the HGE peptide interacted with 48/34-kDa proteins on the membrane of neural stem cells. The periphery cells and rhesus monkey ES cells were used as a negative selection. Combined with quantum dots, the HGEVPRFHAVHL peptide can be used as a direct tool to show optical imaging of specific binding on a single cell membrane. Further results of Western blot showed that the HGEVPRFHAVHL peptide interacted with 48/34-kDa proteins on the membrane of neural stem cells. 1517 AETVESCLAKSH(9) ALTLHPQPLDHP(4) QNMMSPIEGVRI(1) APRYTQTPQALA(1) FMGPQESTLQRL(1) TALATSSTYDPH(1) KSWLPLSQEVRF(1) 7 Phage display (common panning) 6 PMID:20350526 Bacillus cereus ATCC 4342 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The method enabled us to identify two 12-amino acid consensus peptide sequences (AETVESCLAKSH and ALTLHPQPLDHP) that specifically bind to B. cereus 4342 and B. anthracis Sterne, the nonpathogenic surrogates of B. anthracis strain. 1518 DSLQASAT[100, 17] ADLTVQAN[97, 2] AELTTRAE[94, 3] DQLNATAL[113, 1] DQLTVSAQ[84, 18] DSLHGQAM[157, 1] DSLTLQAQ[79, 3] DTLTHEAT[63, 2] EDLTQRAL[66, 3] SNLEMMAT[100, 1] DDLTASAI[1, 130] DELITEAH[<1, 160] DELTVAAN[0, 142] DLLTTQAE[<1, 94] DNLEMMAQ[1, 104] DNLITMAD[<1, 105] DSLNEQAV[1, 138] DTLTENAV[0, 160] EDLNAQAL[1, 113] EDLSAMAG[1, 115] EELESIAN[<1, 129] EELNGQAM[1, 130] EELNQQAN[1, 104] EELSNSAT[2, 123] EELSQQAN[1, 153] EELSVQAT[1, 107] EELTLEAH[1, 146] EELTNSAQ[<1, 168] ELLEAQAK[2, 94] ELLTDQAS[<1, 122] ENLNEVAM[1, 130] EQLENQAI[2, 123] EQLEVQAS[1, 120] EQLSSEAN[1, 179] EQLSSNAE[0, 132] EQLTVEAS[2, 113] ESLAAEAT[1, 152] ESLEAQAT[1, 137] ESLTENAY[<1, 133] ETLENMAS[<1, 97] ETLEQQAT[1, 114] ETLEVQAQ[2, 111] ETLTESAA[1, 148] LDLHEAAV[<1, 135] QELSIQAE[1, 119] SLLSNQAE[1, 131] 46 Phage display (common panning) 3 PMID:19633313 β-galactosidase Not determined. Not determined. Not determined. G-α phage display library (XXLXXXAX) ELISA ELISA signals are normalized against the 1G40 phage (displaying DSLQASAT) ELISA signal in direct ELISA. ELISA signals are normalized against the signal of the sample without phage in competitive ELISA. Both normalized ELISA signals were shown. Normalized ELISA signals of control phage in the direct format and in the competition format were <1 and 115, respectively. 1519 AYLADRAD(34) FDLQLLAE(4) AYLLLRAD(1) 3 Phage display (common panning) 3 PMID:12538914 Bovine fibrinogen Not determined. Not determined. Not determined. f8 α phage display library (XXLXXXAX) 1520 EAGPRAAP(3) EAGPRSTP(3) EAGPRSSP(2) EAGPRSAP(1) EAGPRSNP(1) AAGPRPTS(1) EAGPRSTQ(1) VAGPREVP(1) VAGPRMTE(1) VAGPREVS(1) EAGPRASP(1) AGPREPNL(1) EAGPRSQP(1) EGYLRPDT(1) DGFLRPE(1) EGYLRPES(1) EGYMRPDP(1) DGFLRPDP(1) DGFLRPDS(1) EGFTRPSP(1) DSSVRFTG(1) AGGPRTAP(1) 22 Phage display (common panning) 3 PMID:12538914 Bovine fibrinogen Not determined. Not determined. Not determined. f8-8mer phage display library (X8) 1521 HGHPYQHLLRVL DMPRTTMSPPPR 2 Phage display (common panning) PMID:19342549 Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1522 ANPHTTLRNHVL(1) SPRLTMPPKVPG(1) HPSLNLPLPRLV(1) DLNTYPTLSKRE(1) VSSYPPPQTLLP(1) TQNSNSTQYGNR(1) DRPPHLGDPPHM(1) 7 Phage display (common panning) 3 DOI:10.1002/adma.200702322 Hydroxyapatite, HA Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) This step was necessary to remove any phage with strong affinity to the polypropylene filter tube. The library was then used to perform the subsequent selection against hydroxyapatite (HA). 1523 QHTNIVNTQSRV(1) KLPPINLHPHRL(1) TAPASMSDDRAS(1) SILSTMSPHGAT(1) SSPDRALAATPF(1) LLADTTHHRPWT(1) 6 Phage display (common panning) 5 DOI:10.1002/adma.200702322 Hydroxyapatite, HA Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) This step was necessary to remove any phage with strong affinity to the polypropylene filter tube. The library was then used to perform the subsequent selection against hydroxyapatite (HA). 1524 SVSVGMKPSPRP(19) SVSVGMNPSPRP(1) SAHGTSTGVPWP(1) FPWLPRDNHTLN(1) AVSSLSSTNYSI(1) TMGPTAPRFPHY(1) QSHTRHISPAQV(1) SAKTLSNSPLSN(1) DAQQITLSHWRC(1) 9 Phage display (common panning) 6 DOI:10.1002/adma.200702322 Hydroxyapatite, HA Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) This step was necessary to remove any phage with strong affinity to the polypropylene filter tube. The library was then used to perform the subsequent selection against hydroxyapatite (HA). 1525 CGNSNPKSC(12) CPHNLTKLC(2) 2 Phage display (in vivo) 4 PMID:15492500 Human gastric cancer cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) After four rounds of selection, one phage was obtained with a cyclic 7-mer peptide CGNSNPKSC homing to human gastric adenocarcinoma . There was a 4.6~137.26-fold increase in the number of the selected phage in gastric cancer xenograft in comparision with control organs brain, heart, liver, spleen and kidney. Immunohistochemistry in mouse and human tissue showed that this phage peptide only bind to the endothelial cells of human gastric cancer. This peptide was observed only specific binding to HUVEC not to SGC-7901, Eca-109, LoVo and Hep-G2 by ELISA. The competitive and inhibitory result between the synthetic CGNSNPKSC peptide and the phage displaying the peptide CGNSNPKSC on HUVEC and in vivo was also confirmed its specific binding effect. This peptide may be a possible candidate for targeted drug delivery in antivascular therapy. 1526 DMPGTVLP 1 Phage display (common panning) 4 PMID:21050894 Human breast cancer cell line MCF-7 Not determined. Not determined. Not determined. f8-8mer phage display library (X8) The phage DMPGTVLP (designated by the structure of the borne foreign peptide) demonstrates high selectivity and specificity towards target cells versus control unrelated cells. 1527 AHSLKSITNHGL(1) GLNLPQNKVSFS(1) GPSYLHRLVPAF(1) LPDPHATNILFR(1) SEAVRHLAGPPR(1) SISTSFHAYKLK(1) TGNYKALHPHNG(12) TPQLPIDVNADR(1) YLPDQQLTWFPS(1) 9 Phage display (in vivo) 4 PMID:21470674 ICR male mice brain Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage Clone displaying Pep TGNYKALHPHNG revealed a significant superiority on brain transport efficiency compared with native M13 phage. When conjugated on the surface of PLGA nanoparticles, Pep TGNYKALHPHNG facilitated the targeted delivery of nanoparticles across the BBB, leading to significant higher bEnd.3 cells uptake and in vivo brain accumulation. 1528 NLLNHPQ(2) SLLAHPQ(1) STHTSAQ(1) SLIAHPQ(1) TLLAHPQ(1) HPQ 5 Phage display (common panning) 3 PMID:21473586 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. Ph.D.-7 phage display library (X7) ELISA,Surface plasmon resonance (SPR) The peptide SLLAHPQGGG-mpal was coupled via NCL to the fluorescein labeled EG linker and purified by prep-HPLC (yield = 60%). ELISA and Biacore measurements were performed. The ELISA (on a 100% SA packed The peptide SLLAHPQGGG-mpal was coupled via NCL to the fluorescein labeled EG linker and purified by prep-HPLC (yield = 60%). ELISA and Biacore measurements were performed. The ELISA (on a 100% SA packed\r\nsurface) showed a Kd of 97 μM, while the Biacore measurements yielded 57 μM. 1529 AMYYPLWPSLVY(1) APGYARLPSLMS(1) AVGGQTPIRAKI(1) AYSPISTVTQPY(1) CPLPYPLCLPHG(1) DAMIMKKHWHRF(1) DLQALLPNYPRI(1) ELTWPHVHRKPS(1) ERQGTPYSMYVL(1) FHKEWRTHFQQR(1) FHKHSPRSPIFI(2) FHKYPRPVAMTF(1) FPKAFHHHKIYK(1) GLLHHKHHRSPY(1) GPLLVLNSHSFD(1) GSAVASTLPLGQ(1) HFKHQHSYARPP(1) HFKYHHHMLRSP(1) HFNHTIPLSANP(1) HGSKHMPQQSTH(1) HILPWKIPAHSA(1) HILSPSGSPRMS(1) HISPISAYPWVS(2) HLPRHHWQWPSR(1) HSFHSHVHLKNR(1) HSYMPPLPPQLY(1) IPHPHRWPLHSH(1) IPTMPHPSTARE(1) IQSGTPHPPLRS(1) LEAPRPTPAVPM(1) LHSSVQLTYPLP(1) LITNNPGRLPPQ(1) LLHAPYDHSVSP(1) LSPLYPQLLGLA(3) LTHNKMHREQAS(1) LTYQPLFPTLVY(1) NAISWFPMHLAH(1) NISSIRPTLVEV(1) NVHIRQPLGASS(1) QHVQHQLASTGE(1) QNNLDYIGLYAR(1) QSMLGFISVSAK(1) QTTTAPRHTLWL(1) SAPYKPLLHHFG(1) SHEIYVGSDGFR(1) SHWWARVPFYPP(1) SLVPSYHRSLST(1) SMMMPDQLSLGR(1) SMTHLYTDLWQP(1) SPPTPHHPHPRL(1) SPRTPDLTSLLP(1) SSPFXWQSFSEV(1) STAQPRFGPSSL(1) SYSRTVPPAQWP(1) TCPRYICQAPHP(4) TLTWHTKTPVRP(1) TNVPNPLQPNPR(1) TTGLVQPTAIDP(1) TTNIYFNTPAEV(1) WHKHIPSPRASS(1) WHKHPHAVFNAR(1) WHQSWWAARLGQ(1) WHSSWKSRVVPT(1) YALKHLPESTIP(1) YKAPRHLASHLF(1) YPTSNIIPSIWS(1) 66 Phage display (common panning) 3 PMID:21496351 AC3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In fact, the target is purified recombinant AC3 protein with Maltose Binding Protein as fusion tag (MBP-AC3). Few of these interacting peptides were found to be homologous to proteins from replication process, RNAi pathway, histone and DNA modifying enzymes indicating the role of AC3 in these pathways. 1530 TPITQLL(10) QTSSAAL(4) STFTTTL(4) SLPLPKP(2) 4 Phage display (common panning) 3 PMID:21533529 The first and second extracellular loops of CCR5 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) These peptides could significantly protect against and reduce the severity of EAE. The infiltration of monocytes and lymphocytes into the spinal cord decreased significantly in treated mice, while abundant infiammatory cells and demyelination were observed in spinal cords of EAE mice. 1531 RRSHPCRTCTTHTP HRKTTCTRCPATSP HRRGECRACPLLPA RRPAHCHHCPRNPL 4 Phage display (common panning) 3 PMID:21539876 Anti-PSA monoclonal antibody Prostate-specific antigen, PSA Not determined. Not determined. f88-Cys2 phage display library (X5CX2CX5) Western blot In Western blot, peptide 1 (RRSHPCRTCTTHTP) antiserum recognized PSA protein where as peptides 2 (HRKTTCTRCPATSP), 3 (HRRGECRACPLLPA) and 4 (RRPAHCHHCPRNPL) does not. These peptides apart from differences in their amino acid sequence, elicited minimal cross reactive antibody responses against each other. One of the four peptides analyzed produced an antibody response that recognizes the PSA protein. 1532 AQSPTIKLTPSW(1) ANSYSVNYTPSM(1) SVSVGMKPSPRP(7) ESSYSWSPARLS(1) KPFHDWLYSPTA(1) ATPHYTVGQWNQ(3) NHNMHRTTQWPL(1) LPNIPYHHPLYH(1) SDIRNWDTQSLP(2) SDIWPLQSQMYP(1) 10 Phage display (common panning) 3 PMID:21544611 Human ovarian tumor cell line SK-OV-3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Cells were washed twice with serum-free medium, and re-incubated in serum-free medium for 2 h at 37 ℃ to clear their surface receptors.', '3', '21544611', 'The synthetic biotin-labeled peptide, SVSVGMKPSPRP, demonstrated a high specificity to SK-OV-3 cells especially when compared to other cell lines (A2780 and 3T3). The synthetic biotin-labeled peptide, SVSVGMKPSPRP, demonstrated a high specificity to SK-OV-3 cells especially when compared to other cell lines (A2780 and 3T3). 1533 STLPLPP(9) SPMTLYG(6) TMQMTRY(6) LVTTGPL(4) QTHSWWP(2) 5 Phage display (competitive panning) 3 PMID:21567665 Lead Ion, Pb(2+) Imidazol Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted with 200 mM imidazol solution. Isothermal calorimetric analysis revealed that the peptide STLPLPP bound to Pb(2+), Cd(2+), Hg(2+), and Cu(2+). Through the use of CD studies, no secondary structural changes were observed for the peptide upon binding to divalent cations. Ala scanning mutant peptides bound to Hg(2+) with a reduced affinity. However, no single substitution was shown to affect the overall affinity. 1534 GRQYYEGRKPDYRAA(18) 1 Phage display (common panning) 2 PMID:21573118 Anti-HMW-MAA monoclonal antibody VT80.12 High molecular weight melanoma-mssociated antigen, HMW-MAA Not determined. Not determined. X15 PC89 phage diaplay library ELISA In ELISA inhibition experiments, the peptide GRQYYEGRKPDYRAAC inhibited the binding of the mAb VT80.12 to the HMW-MAA up to 93%. Twenty individual phage clones were tested for their ligand specificity in phage ELISA. Among these, eighteen phage clones specifically bound to the mAb VT80.12. DNA sequencing of these 18 phage clones yielded one peptide sequence GRQYYEGRKPDYRAA. Peptide inhibited the binding of mAb VT80.12 to the HMW-MAA. 1535 GRQYYEGRKPDYRAA(19) 1 Phage display (common panning) 3 PMID:21573118 Anti-HMW-MAA monoclonal antibody VT80.12 High molecular weight melanoma-mssociated antigen, HMW-MAA Not determined. Not determined. X15 PC89 phage diaplay library ELISA In ELISA inhibition experiments, the peptide GRQYYEGRKPDYRAAC inhibited the binding of the mAb VT80.12 to the HMW-MAA up to 93%. Twenty individual phage clones were tested for their ligand specificity in phage ELISA. Among these, nineteen phage clones specifically bound to the mAb VT80.12. DNA sequencing of these 19 phage clones yielded one peptide sequence GRQYYEGRKPDYRAA. Peptide inhibited the binding of mAb VT80.12 to the HMW-MAA. 1536 NYQDLQRTHFKS(20) 1 Phage display (common panning) 3 PMID:21573118 Anti-HMW-MAA monoclonal antibody VF1-TP43 High molecular weight melanoma-mssociated antigen, HMW-MAA Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA In ELISA inhibition experiments, the peptide NYQDLQRTHFKSGPGPGC inhibited the binding of the mAb VF1-TP43 to the HMW-MAA up to 100%. Among 30 tested phage clones, 20 were specifically recognized by the mAb VF1-TP43 in phage ELISA. DNA sequencing yielded one peptide sequence NYQDLQRTHFKS. Peptide inhibited the binding of mAb VF1-TP43 to the HMW-MAA. 1537 WHWTYYW(39) THKFPWI(1) WHWLWLQ(1) HLPPNHT(1) KLWTIPM(1) SSLRLLP(1) 6 Phage display (common panning) 3 PMID:21592685 Major outer membrane protein Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) To identify host proteins that interact with LipL32, phage display technology was employed in the study. The nucleotide and corresponding peptide sequences from pyrosequencing were analyzed and used to search for matched host proteins in the protein databases using the BLASTP program of National Center for Biotechnology Information (NCBI). Putative proteins with potential binding to LipL32 are proteins known to be expressed on the surface of target cells of pathogenic Leptospira such as chloride channel accessory 2, glycoprotein VI, scavenger receptor expressed by endothelial cell isoform I (SREC-I), coronin 2A, laminin alpha 5, collagen XX, and prostaglandin receptor EP1. 1538 IYSNTSAPHLII(2)[1.294 ± 0.245/1.754 ± 0.258] LTRGYNTSLFQE(1)[1.986 ± 0.189] GHTLNAPINLPM(1)[1.621 ± 0.094] ALHMKLGPQYYP(1)[1.513 ± 0.139] LTNQLQHTWISP(1)[3.308 ± 0.303] SATMGRVPTTAS(1)[1.318 ± 0.126] YHETRIIEGADS(1)[1.268 ± 0.120] NTPSPYPMSSAP(1)[1.558 ± 0.107] VFRDVTPMLLTY(1)[2.648 ± 0.037] MNYMNITISVEK(1)[1.891 ± 0.132] KIMPPNWLVAKL(1)[2.270 ± 0.258] NFTSHDFWFWMT(1)[2.824 ± 0.283] NDPNNVNPTAGR(1)[1.513 ± 0.227] KYLAYPDSVHIW(1)[3.371 ± 0.202] 14 Phage display (subtractive panning) 3-4 PMID:21611873 LNCaP prostate carcinoma cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Binding of the selected phages on LNCaP and PC-3 cells was examined with cell phage ELISA. OD450(LNCap)/OD450(PC-3) was reproduced from the graph and shown. Results are represented as the mean ± SD (n=3). The value of the insertless phage was 0.979 ± 0.397. Biopanning were performed on LNCaP cells after pre-cleaning on PC-3 cells to remove the non-specific bound phages. Cell phage ELISA and immunostaining demonstrated high specificity of phage clone (KYLAYPDSVHIW) to LNCaP cells. 1539 MPKDMLPYANPN QSFKDHLPAGMR HSFEPWSARDML NMSLRDHYSPSP WSPSKLYSARDH THRPLSHNFRDF 6 Phage display (subtractive panning, competitive panning) 3 PMID:21695105 Anti-VSG LiTat 1.5 monoclonal antibody H12H3 Variable surface glycoprotein LiTat 1.5 Not determined. Not determined. Ph.D.-12 phage display library (X12) Pannings were performed in the first two rounds consisting of 1) a positive selection with anti-VSG mAbs coated on magnetic particles (MP), 2) a negative selection with anti-VSG mAb-free MP. Bound phages were eluted via antigen competition. 1540 ALLPFKDHLPYP APQNAKDHLPGY APWSLRDHLAVT FAENKKDHFSPG QPNHRDHFKLPA TPGVERDHFVRL SLMHVYRDHSTS SNWRDHYGLGAG MQDSRLRLALSA 9 Phage display (subtractive panning) 3 PMID:21695105 Anti-VSG LiTat 1.5 monoclonal antibody H12H3 Variable surface glycoprotein LiTat 1.5 Not determined. Not determined. Ph.D.-12 phage display library (X12) Pannings were performed in the first two rounds consisting of 1) a positive selection with anti-VSG mAbs coated on magnetic particles (MP), 2) a negative selection with anti-VSG mAb-free MP. 1541 CWPSMRDHC CTYSPRDMC CRDHFSTHC CRDHYPLSC CKDHATDLC CISPNQWLC CTSPNQWLC CLDAKLQQC CPNFELHWC CPPYETWWC CNVYEAYWC CTDFEGMLC 12 Phage display (subtractive panning) 3 PMID:21695105 Anti-VSG LiTat 1.5 monoclonal antibody H12H3 Variable surface glycoprotein LiTat 1.5 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Pannings were performed in the first two rounds consisting of 1) a positive selection with anti-VSG mAbs coated on magnetic particles (MP), 2) a negative selection with anti-VSG mAb-free MP. 1542 CWYSTRDHC CRDHYDQMC CRDHSDLWC CRDHLYGSC CKDHATDLC CWALRDPLC CNALNQWLC CRTSNQWLC CTALQQWLC CLDAKLQQC CQMPHWWHC CQVPHFFGC 12 Phage display (subtractive panning, competitive panning) 3 PMID:21695105 Anti-VSG LiTat 1.5 monoclonal antibody H12H3 Variable surface glycoprotein LiTat 1.5 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Pannings were performed in the first two rounds consisting of 1) a positive selection with anti-VSG mAbs coated on magnetic particles (MP), 2) a negative selection with anti-VSG mAb-free MP. Bound phages were eluted via antigen competition. 1543 MDWINPFPNFNS RVPNVLPLFPFL NSPFMLHMSALS 3 Phage display (subtractive panning) 3 PMID:21695105 Anti-VSG LiTat 1.3 monoclonal antibody H13F7 VSG protein Not determined. Not determined. Ph.D.-12 phage display library (X12) Pannings were performed in the first two rounds consisting of 1) a positive selection with anti-VSG mAbs coated on magnetic particles (MP), 2) a negative selection with anti-VSG mAb-free MP. 1544 QPAIWINPFPAW TPSWRNPFPTFY LTPPPWQNPFPP AHPPWPLQYVFR TPTWPLLTIFPS NSPFMLHMSALS SELSPAMLHFMF 7 Phage display (subtractive panning, competitive panning) 3 PMID:21695105 Anti-VSG LiTat 1.3 monoclonal antibody H13F7 VSG protein Not determined. Not determined. Ph.D.-12 phage display library (X12) Pannings were performed in the first two rounds consisting of 1) a positive selection with anti-VSG mAbs coated on magnetic particles (MP), 2) a negative selection with anti-VSG mAb-free MP. Bound phages were eluted via antigen competition. 1545 CWLPLTRLC CWHPLQWRC CLEILSWLC CNQVLSWLC CILPWKHIC CVLPWSHVC CFLPHPSRC 7 Phage display (subtractive panning, competitive panning) 3 PMID:21695105 Anti-VSG LiTat 1.3 monoclonal antibody H13F7 VSG protein Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Pannings were performed in the first two rounds consisting of 1) a positive selection with anti-VSG mAbs coated on magnetic particles (MP), 2) a negative selection with anti-VSG mAb-free MP. Bound phages were eluted via antigen competition. 1546 SHSTPYYWKGYI 1 Phage display (subtractive panning, competitive panning) 3 PMID:21695105 Anti-VSG LiTat 1.3 monoclonal antibody H18C11 VSG protein Not determined. Not determined. Ph.D.-12 phage display library (X12) Pannings were performed in the first two rounds consisting of 1) a positive selection with anti-VSG mAbs coated on magnetic particles (MP), 2) a negative selection with anti-VSG mAb-free MP. Bound phages were eluted via antigen competition. 1547 FHWSWYTPSRPS(2)[0.579/0.354] WHWPWYQGQLWP(2)[0.336/0.270] SHWIDWLYSSPI(1)[0.169] QHWYLWNLMYGA(1)[0.428] WHWQYTPWWRGS(1)[0.610] WHWQWTPWSIQP(1)[0.480] FHWTQYFSPWIR(1)[0.597] WHWSLYPLTYPP(1)[0.318] [AVLIFYW]-H-W-x-[AVLIFYW]-[AVLIFYW]-x-[AVLIFYW]-x-[AVLIFYW] 8 Phage display (competitive panning) 3 PMID:21713010 Human antimicrobial peptide LL-37 Glucan 1,3-beta-glucosidase Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA In phage ELISA, the absorbance at 450 nm was determined using an ELISA reader. Data shown were reproduced from the graph. LL-37 samples were added to the wells and incubated temperature to elute the bound phage. Using phage display and ELISA, authors identified 10 peptide sequences that could bind LL-37. A BLAST search revealed that four sequences in the major C. albicans cell-wall b-1,3-exoglucanase, Xog1p, were highly similar to the consensus sequence derived from the 10 biopanned peptides. One Xog1p-derived peptide, Xog1p 90-115, and recombinant Xog1p associated with LL-37, thereby reversing the inhibitory effect of LL-37 on C. albicans adhesion. LL-37 reduced Xog1p activity and thus interrupted cell-wall remodeling. 1548 STTPQSVYASAP(1) AVPLNTSVLHTT(1) PMPSNRPGYNVS(1) YGCNTTPCHWAM(1) GRLDDHNQSRML(1) NTSHEILWQTYP(1) SHGTLQRVHTWS(1) DWRVIIPPRPSA(2) AAFTPRPWPSTV(1) YDESTYHQGQKR(1) 10 Phage display (subtractive panning) 1 PMID:21624651 Rabbit chondrocyte Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the negative selection procedure, the phage display library was incubated with the synovial tissue and fluid to exclude the synovial affinity phage clones. Analysis suggests that the peptide DWRVIIPPRPSA can efficiently interact specifically with chondrocytes without any species specificity. Polyethylenimine (PEI) was covalently modified with CAP to construct a non-viral vector for cartilage-targeted therapy. 1549 DWRVIIPPRPSA(12) 10 Phage display (subtractive panning) 2 PMID:21624651 Rabbit chondrocyte Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the negative selection procedure, the phage display library was incubated with the synovial tissue and fluid to exclude the synovial affinity phage clones. Analysis suggests that the peptide DWRVIIPPRPSA can efficiently interact specifically with chondrocytes without any species specificity. Polyethylenimine (PEI) was covalently modified with CAP to construct a non-viral vector for cartilage-targeted therapy. 1550 LRSRTKIIRIRH 1 Phage display (common panning) 3 PMID:21596749 Heparan sulfate, HS Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Viral entry and glycoprotein D binding assays together with fluorescent microscopy data indicated that peptide LRSRTKIIRIRH was potent in blocking HSV-1 entry into primary cultures of human corneal fibroblasts and CHO-K1 cells transiently expressing different glycoprotein D receptors. 1551 MPRRRRIRRRQK 1 Phage display (common panning) 3 PMID:21596749 3-O-sulfated heparan sulfate, 3-OS HS Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Viral entry and glycoprotein D binding assays together with fluorescent microscopy data indicated that peptide MPRRRRIRRRQK was potent in blocking HSV-1 entry into primary cultures of human corneal fibroblasts and CHO-K1 cells transiently expressing different glycoprotein D receptors. Interestingly, peptide MPRRRRIRRRQK isolated against 3-OS HS displayed wider ability to inhibit entry of clinically relevant strains of HSV-1 and some divergent members of herpesvirus family including cytomegalovirus and human herpesvirus-8. 1552 CKHYGGGVAC CYKNVDSGGC CRFLLPQGC CYEGSEVSC CLRQGNPTC CGSGGMSPSC CSWKYWFGEC CGQWLGNWLC 8 Phage display (in vivo) 4 PMID:21683670 Mouse adipose stromal cells, ASCs Not determined. Not determined. Not determined. CX7C, CX8C, and CX9C FUSE5-based phage display library pool Comparative analysis of individual phage clones in vivo revealed peptide CSWKYWFGEC, homing to ASCs 1000 times more efficiently than to LSC, as a peptide with the highest specificity for ASCs. 1553 CAVYRSTGC CESGFPTVGC CLGVGPGFC CIRGKAGRC 4 Phage display (in vivo) 4 PMID:21683670 Mouse lung stromal cells, LSCs Not determined. Not determined. Not determined. CX7C, CX8C, and CX9C FUSE5-based phage display library pool 1554 NSSASSRGNSSSNSVY NSLRKYSKLK 2 Phage display (common panning) 4 PMID:21610657 Etoposide, VP-16 Transcription factor E2F4, E2F-4 Not determined. Not determined. T7 phage display library Biotinylated etoposide derivative was immobilized on a streptavidin-coated 96-well microplate. Etoposide (VP-16) is an anti-tumor compound that targets topoisomerase II (top II). In this study, authours had identified an alternative binding protein of etoposide by screening a library of T7 phage-displayed peptides. Peptide NSSASSRGNSSSNSVY displays similarity with the ser-rich domain in E2F-4, a transcription factor in cell cycle-regulated genes, suggesting that etoposide might interact with E2F-4 via this domain. 1555 EAHVMHKVAPRP QNTATAVSRLSP ATHTNQTHALYR VSNHKALDYPTR DSGRYSMTNHYS 5 Phage display (common panning) 3 DOI:10.1002/adma.200500863 Zinc oxide, ZnO Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Five different sequences were determined from 35 clones. Approximately half of the selected phage viruses (18 of 35) displayed the same ZnO-1 peptide (EAHVMHKVAPRP). Comparison of the sequences of the selected peptides indicates a preponderance of amino acid residues with functional side chains. Interestingly, the ZnO-1 peptide has a greater number of basic and hydrophobic residues than the other selected peptides. The binding affinity for the monoclonal phage displaying each selected ZnO-binding peptide was estimated from the ratio of the number of bound phage viruses to the total number of phage viruses added to the ZnO-binding assay. 1556 SVSVGMKPSPRP(4)[NT] TMGFTRPRFPHY(3)[NT] SWFSYLSPWVRS(2)[NT] TWDWRMFPLRPW(1)[NT] MYLICPEQRSPL(1)[0.67] HPSVIPPSPVMS(1)[NT] HYLIGLGCIGLN(1)[NT] HKAHVSKHSVPI(1)[NT] GPLPSNHSFQHP(1)[NT] 9 Phage display (common panning) 5 PMID:10219262 CSF samples from neurocysticercosis patient #1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA In phage ELISA, the absorbance at 450 nm was measured. Data shown are expressed as the difference between the average A405nm values for selected phage and nonrelated phage G1. NT represents not tested. To increase the specificity of selection, plates coated with anti-human Fc antibody were used in five rounds of biopanning. Sequence similarities between affinity-selected clones and three known T. solium and T. crassiceps proteins were encountered. 1557 SHVPRIGGPNFW(2)[NT] FSVPWPTRPPHW(2)[1.28] SWNHWLYTYFPQ(2)[NT] WPYKPYWMPNFW(2)[NT] SIDWWHLIYPRP(1)[NT] SWNHWLYSGART(1)[NT] SLFRRRTSTPHR(1)[NT] SWWPFPPQPDPA(1)[NT] NTNPAWRHYKAF(1)[NT] SDNVHTWQAMFK(1)[NT] AYYKTSFPPSTQ(1)[NT] 11 Phage display (common panning) 5 PMID:10219262 CSF samples from neurocysticercosis patient #2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA In phage ELISA, the absorbance at 450 nm was measured. Data shown are expressed as the difference between the average A405nm values for selected phage and nonrelated phage G1. NT represents not tested. To increase the specificity of selection, plates coated with anti-human Fc antibody were used in five rounds of biopanning. Sequence similarities between affinity-selected clones and three known T. solium and T. crassiceps proteins were encountered. 1558 VPHIPPN(1) MPPTQVS(1) QMHPWPP(1) QPPFWQF(1) TPPQGLA(1) IPPYNTL(1) AVRPAPL(1) GAKPHPQ(1) QQLSPLP(1) GPPPSPV(1) LPLTPLP(1) 11 Phage display (competitive panning) 3 PMID:11157741 Melanoma-derived growth regulatory protein Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding recombinant human MIA at a higher concentration. 1559 QLNVNHQARADQ(1) TSASTRPELHYP(1) TFLPHQMHPWPP(1) VPHIPPNSMALT(1) RLTLLVLIMPAP(5) 5 Phage display (competitive panning) 3 PMID:11157741 Melanoma-derived growth regulatory protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were eluted by adding recombinant human MIA at a higher concentration. 1560 SVSVGMKPSPRP(3)[0.313 ± 0.004, 25.0%] YAPSSLLPHSVD(2)[0.236 ± 0.006, 29.2%] NSQLRLNYEKSL(2)[0.494 ± 0.007, 29.5%] MPDHALRPQYSI(1)[0.098 ± 0.006, 13.4%] HLKYATYPPYPQ(1)[0.373 ± 0.010, 35.2%] HNYYNKIQTAAH(1)[0.051 ± 0.008, 16.7%] 6 Phage display (common panning) 3 PMID:11376846 Anti-JEV E protein monoclonal antibody E3.3 Envelope protein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA ELISA product was determined as A405mAb E3.3 - A405BSA. The ratio of inhibition was determined as 1 - (A405phage)/(A405no phage). M13KO7 phage, wild-type phage acted as negative control. Its absorbance and inhibition ratio were 0.006 ± 0.005 and 11.8%, respectively. The specific peptide ligands presented on ten high-affinity phage clones displayed six different amino acid sequences, all showing a novel cis-proline turn structure. After being superimposed onto the best fit of the three-dimensional structure of JEV E protein, these peptide structures were mapped to a conformational region constituted by three continuous polypeptide segments (E307-E309, E327-E333, E386-E390) in domain III. 1561 SVSVGMKPSPRP(9) 1 Phage display (competitive panning) 8 PMID:11562993 Deoxyribonucleotides, DNA Modification methylase TaqI, M.TaqI Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA In ELISA, the selected phage (SVSVGMKPSPRP) led to an UV absorption increase of 30 mOD/min at 405 nm, while the control phage from the primary library yielded only an increase of 5 mOD/min. For competitive elution using the DNA methyltransferase M.TaqI in the selection step, a biotin-labeled duplex oligodeoxyribonucleotide containing the 5'-TCGA-3' recognition sequence of M.TaqI was employed. Biotinylated DNA-phage complexes were captured with streptavidin-coated paramagnetic beads. The N-terminal amino acid sequence SVSVGMKPSPRP was identified in nine out of ten phages sequenced after eight rounds of selection. 1562 PPPLYF RFCDTS RSRLIW 3 Phage display (common panning) 6 PMID:7980552 Single stranded oligonucleotides consisting of cytosine Not determined. Not determined. Not determined. f3-6mer phage display library (X6) Selection were performed in 50 mM 2[N-Morpholino]ethanesulfonic acid(MES)-buffer. Biotinylated DNA-phage complexes were captured with streptavidin-coated agarose beads. The peptides interacted more strongly with oligo-C compared to the original peptide phage library and wildtype phage. The selected clones also showed different specificity in the interaction with oligo-C, -G, -A and -T. 1563 CSWLHQPYC(13)[23.18 ± 1.16][10.90 ± 0.95] CSWFHMPYC(3)[NT][NT] CSWDHMPYC(2)[NT][NT] CRDYAMPLC(2)[61.87 ± 1.26][36.25 ± 6.85] CKTDPWDYC(2)[NT][NT] CSWVHMPYC(1)[NT][NT] CSWMHMPYC(1)[105.2 ± 1.56][184.36 ± 18.45] CNTIGGYEC(1)[9.64 ± 1.15][23.73 ± 2.09] CLTIGGYEC(1)[NT][NT] CRTLGGYEC(1)[NT][NT] S-W-x-H-[MQ]-P-Y, x-T-[LI]-G(2)-Y-E 10 Phage display (common panning) 3 PMID:11923297 Anti-LOS monoclonal antibody 9-2-L379 L3,7,9 lipooligosaccharide, LOS Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA,Resonant mirror biosensor The antigenicity of the enriched phage clones against mAb 9-2-L379 was tested by ELISA. Absorbance was measured at 450 nm (data not shown). Phage clones expressing all the cyclic peptides reacted strongly with mAb 9-2-L379. To determine whether the anti-LOS mAb 9-2-L379 interacted with the consensus cyclic peptides by the same paratope as it interacts with LOS, insolution competitive inhibition ELISAs against meningococcal LOS L3,7,9 were performed. Data were shown and expressed as EC50 ± S.E. (μM). No inhibition (NI) was observed for P-CRC, which is a negative clone. Besides, real time binding kinetics between mAb 9-2-L379 and cyclic peptides were measured with a resonant mirror biosensor. The binding affinity is expressed as the equilibrium dissociation constant (KD, nM), calculated from the ratio of Kd/Ka. The KD for LOS is 7.52 ± 3.8 nM. 1564 CSWVHMPYC(1) 1 Phage display (common panning) 4 PMID:11923297 Anti-LOS monoclonal antibody 9-2-L379 L3,7,9 lipooligosaccharide, LOS Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The fourth round of panning was performed at 50 μg/ml of the target antibody 9-2-L379 coating the microtiter plate wells. 1565 CSWLHMPYC(4)[26.12 ± 1.12][18.76 ± 2.63] CSWDHNPYC(2)[NT][NT] 2 Phage display (common panning) 4 PMID:11923297 Anti-LOS monoclonal antibody 9-2-L379 L3,7,9 lipooligosaccharide, LOS Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA,Resonant mirror biosensor The antigenicity of the enriched phage clones against mAb 9-2-L379 was tested by ELISA. Absorbance was measured at 450 nm (data not shown). Phage clones expressing all the cyclic peptides reacted strongly with mAb 9-2-L379. To determine whether the anti-LOS mAb 9-2-L379 interacted with the consensus cyclic peptides by the same paratope as it interacts with LOS, insolution competitive inhibition ELISAs against meningococcal LOS L3,7,9 were performed. Data were shown and expressed as EC50 ± S.E. (μM). No inhibition (NI) was observed for P-CRC, which is a negative clone. Besides, real time binding kinetics between mAb 9-2-L379 and cyclic peptides were measured with a resonant mirror biosensor. The binding affinity is expressed as the equilibrium dissociation constant (KD, nM), calculated from the ratio of Kd/Ka. The KD for LOS is 7.52 ± 3.8 nM. The fourth round of panning was performed at 5 μg/ml of the target antibody 9-2-L379 coating the microtiter plate wells. 1566 CSWLHQPYC(18)[23.18 ± 1.16][10.90 ± 0.95] CSWMHMPYC(4)[105.2 ± 1.56][184.36 ± 18.45] CSWFHMPYC(3)[NT][NT] CNTIGGYEC(2)[9.64 ± 1.15][23.73 ± 2.09] S-W-x-H-[MQ]-P-Y, x-T-[LI]-G(2)-Y-E 4 Phage display (common panning) 4 PMID:11923297 Anti-LOS monoclonal antibody 9-2-L379 L3,7,9 lipooligosaccharide, LOS Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA,Resonant mirror biosensor The antigenicity of the enriched phage clones against mAb 9-2-L379 was tested by ELISA. Absorbance was measured at 450 nm (data not shown). Phage clones expressing all the cyclic peptides reacted strongly with mAb 9-2-L379. To determine whether the anti-LOS mAb 9-2-L379 interacted with the consensus cyclic peptides by the same paratope as it interacts with LOS, insolution competitive inhibition ELISAs against meningococcal LOS L3,7,9 were performed. Data were shown and expressed as EC50 ± S.E. (μM). No inhibition (NI) was observed for P-CRC, which is a negative clone. Besides, real time binding kinetics between mAb 9-2-L379 and cyclic peptides were measured with a resonant mirror biosensor. The binding affinity is expressed as the equilibrium dissociation constant (KD, nM), calculated from the ratio of Kd/Ka. The KD for LOS is 7.52 ± 3.8 nM. The fourth round of panning was performed at 0.5 μg/ml of the target antibody 9-2-L379 coating the microtiter plate wells. 1567 SEPVAML(9) LPPNPTK(7) NLPRLYE(3) TYWYMTP(2) 4 Phage display (common panning) 4 PMID:11923297 Anti-LOS monoclonal antibody 9-2-L379 L3,7,9 lipooligosaccharide, LOS Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The antigenicity of the enriched phage clones against mAb 9-2-L379 was tested by ELISA. Absorbance was measured at 450 nm (data not shown). Phage clones expressing the identified linear 7-mer consensus peptides showed no reactivity with mAb 9-2-L379. The fourth round of panning was performed at 50 μg/ml of the target antibody 9-2-L379 coating the microtiter plate wells. 1568 GSMSPYVRWYTP(9) KDASSFQMRPLS(2) SVSVGMKPSPRP(2) KAQSPWSNVDAW(2) HSLKHTQMSYSS(2) 5 Phage display (common panning) 4 PMID:11923297 Anti-LOS monoclonal antibody 9-2-L379 L3,7,9 lipooligosaccharide, LOS Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The antigenicity of the enriched phage clones against mAb 9-2-L379 was tested by ELISA. Absorbance was measured at 450 nm (data not shown). Phage clones expressing the identified linear 12-mer consensus peptides showed no reactivity with mAb 9-2-L379. The fourth round of panning was performed at 50 μg/ml of the target antibody 9-2-L379 coating the microtiter plate wells. 1569 KVWFLPEAAQPS(1) KVWQMYWPSGQP(1) TSSALKCCFIQ(1) 3 Phage display (subtractive panning) 3 PMID:15755589 Anti-HIV-1 monoclonal antibdoy 2G12 Human immunodeficiency virus type 1, HIV-1 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA In ELISA, phage clones (displaying KVWFLPEAAQPS, KVWQMYWPSGQP and TSSALKCCFIQ, respectively) bound to 2G12 but not to human IgG. For each round of panning, the library was preadsorbed on BSA. 1570 KCCYYDHSHALS(3) QPTSHRDLRPPI(2) HSLKNSMLTVMA(2) HLHVHLSLSRPL(2) KVWDIRTADNLH(1) KVWDIRYTTPHA(1) EARVFSSKHWIP(1) TMKCCYSNTSPP(1) HLHVHVKFADRQ(1) 9 Phage display (subtractive panning) 5 PMID:15755589 Anti-HIV-1 monoclonal antibdoy 2G12 Human immunodeficiency virus type 1, HIV-1 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA,Competition experiment In ELISA, phage clones bound to 2G12 but not to human IgG. Besides, the participation of carbohydrate binding sites in the peptide binding was demonstrated by inhibition of the binding of 2G12 to phages (KCCYYDHSHALS, KVWDIRYTTPHA and QPTSHRDLRPPI) or HIV gp160 by mannose, fructose, glucose, mannan and α-D methylmannopyranoside. The binding was specifically inhibited by the mannose containing carbohydrates as well as by fructose but not by glucose. For the first three rounds of panning, the library was preadsorbed on BSA and panned on 2G12. Peptides were selected after adsorbing the library selected in the previous step on human IgG and panning once more on 2G12 in the presence of human IgG (rounds 4 and 5). 1571 SVSVGMKPSPRP(6) KLWQLFPPSAVS(1) EARVXSSKHWIP(1) 3 Phage display (subtractive panning, competitive panning) 6 PMID:15755589 Concanavalin-A, Con A Methyl α-D-mannopyranoside Not determined. Not determined. Ph.D.-12 phage display library (X12) For the first three rounds of panning, the library was preadsorbed on BSA and panned on 2G12. The amplified eluent from the third round of panning was adsorbed on human Immunoglobulin G (huIgG). The non-binding phage was used as input for a fifth round of panning on 2G12 carried out in the presence of huIgG in the liquid phase. Peptides were selected after the library from round 5 was panned on Con A and the bound phages eluted by α-D-methylmannopyranoside. 1572 TMGFTAPREPHY(3) IERPLHESVLAT(2) NNYDDISLRARP(2) AIPNKLNVWPPH(1) TGVSWSVAQPSF(1) SQELTQRPYKWH(1) TPSYINLXDFIA(1) GTSTFNSVPVRD(1) KLTFLNYAEVLR(1) 9 Phage display (subtractive panning, competitive panning) 4 PMID:16603084 Hsp70-peptide complexes, Hsp70-PCs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The blocking, binding and washing strategies were carried out as instructed by the manufacturer with the following exceptions. Authors used (a) either 1% Bovine Serum Albumin (BSA) or casein for blocking nonspecific binding alternating these blocking substrates between subsequent rounds of bio-panning to prevent selection of phage recognising the blocking substances, (b) competitive elution with the bait Hsp70-PCs in rounds three and four and (c) bio-panning in solution for rounds two and four, using biotinylated Hsp70-PCs and a streptavidin matrix to prevent selection of plastic-binding phages. In the latter case, 10 μg of Hsp70-PCs were biotinylated using NHS-Biotin according to the manufacturer\'s instructions and incubated with phage particles as recommended by the manufacturer with either of the blocking reagents. The Hsp70-PCs bound phage particles were recovered either through a Streptavidin-agarose (Pierce Chemical Co. ILL) column or Streptavidin-Dynabeads (Dynal Co., Norway) followed by washing and competitive elution as described above. Following each round of bio-panning, the eluted phages were amplified to high titer according to supplier\'s instructions. A subtraction screening using the peptide depleted Hsp70-PC was performed after the third biopanning to remove those phages recognising the Hsp70 portion of the Hsp70-PC bait. The unbound fraction was amplified and used in the fourth round bio-panning. The phage particles from the final fourth round eluate were plated at low density to allow isolation of single phage clones. Three separate bio-panning experiments were performed using Hsp70-PCs as bait. In each case, approximately 400-1000 phages were retained after four rounds of panning. A total of twenty four phage clones were selected at random for further analysis. 1573 SVSVGMKPSPRP(0.46) FHSDWPGXTLTW(0.18) LHAETRSAMHRT(0.09) WKHTSQPPRLIF(0.09) KAXTPVQSASNV(0.09) RTHDNSWNYTSS(0.09) 6 Phage display (subtractive panning, competitive panning) 4 PMID:16603084 Peptide NNYDDISLRARP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The blocking, binding and washing strategies were carried out as instructed by the manufacturer with the following exceptions. Authors used (a) either 1% Bovine Serum Albumin (BSA) or casein for blocking nonspecific binding alternating these blocking substrates between subsequent rounds of bio-panning to prevent selection of phage recognising the blocking substances, (b) competitive elution with the bait peptide in rounds three and four and (c) bio-panning in solution for rounds two and four, using biotinylated peptide and a streptavidin matrix to prevent selection of plastic-binding phages. In the latter case, 10 μg of peptide were biotinylated using NHS-Biotin according to the manufacturer\'s instructions and incubated with phage particles as recommended by the manufacturer with either of the blocking reagents. The peptide bound phage particles were recovered either through a Streptavidin-agarose (Pierce Chemical Co. ILL) column or Streptavidin-Dynabeads (Dynal Co., Norway) followed by washing and competitive elution as described above. Following each round of bio-panning, the eluted phages were amplified to high titer according to supplier\'s instructions. The amplified fraction was used in the fourth round biopanning. The phage particles from the final fourth round eluate were plated at low density to allow isolation of single phage clones. 1574 DSPQNPKTWKYI(1.00) 1 Phage display (subtractive panning, competitive panning) 4 PMID:16603084 Peptide TMGFTAPREPHY Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The specificity of the interaction between the TMGFTAPREPHY and DSPQNPKTWKYI peptides was examined by ELISA. Phages displaying the DSP peptide specifically bind to a synthetic biotinylated TMG peptide. Conversely, phages displaying the TMG peptide bind to a synthetic biotinylated DSP peptide. The blocking, binding and washing strategies were carried out as instructed by the manufacturer with the following exceptions. Authors used (a) either 1% Bovine Serum Albumin (BSA) or casein for blocking nonspecific binding alternating these blocking substrates between subsequent rounds of bio-panning to prevent selection of phage recognising the blocking substances, (b) competitive elution with the bait peptide in rounds three and four and (c) bio-panning in solution for rounds two and four, using biotinylated peptide and a streptavidin matrix to prevent selection of plastic-binding phages. In the latter case, 10 μg of peptide were biotinylated using NHS-Biotin according to the manufacturer\'s instructions and incubated with phage particles as recommended by the manufacturer with either of the blocking reagents. The peptide bound phage particles were recovered either through a Streptavidin-agarose (Pierce Chemical Co. ILL) column or Streptavidin-Dynabeads (Dynal Co., Norway) followed by washing and competitive elution as described above. Following each round of bio-panning, the eluted phages were amplified to high titer according to supplier\'s instructions. The amplified fraction was used in the fourth round biopanning. The phage particles from the final fourth round eluate were plated at low density to allow isolation of single phage clones. 1575 SVSVGMKPSPRP(0.32) GLPPYSPHRLAQ(0.15) NFMESLPRLGMH(0.15) NAQNYSQQAPRP(0.15) HGLHQMSGNTKR(0.15) HPHQPIERQTVQ(0.08) 6 Phage display (subtractive panning, competitive panning) 4 PMID:16603084 Peptide IERPLHESVLAT Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The blocking, binding and washing strategies were carried out as instructed by the manufacturer with the following exceptions. Authors used (a) either 1% Bovine Serum Albumin (BSA) or casein for blocking nonspecific binding alternating these blocking substrates between subsequent rounds of bio-panning to prevent selection of phage recognising the blocking substances, (b) competitive elution with the bait peptide in rounds three and four and (c) bio-panning in solution for rounds two and four, using biotinylated peptide and a streptavidin matrix to prevent selection of plastic-binding phages. In the latter case, 10 μg of peptide were biotinylated using NHS-Biotin according to the manufacturer\'s instructions and incubated with phage particles as recommended by the manufacturer with either of the blocking reagents. The peptide bound phage particles were recovered either through a Streptavidin-agarose (Pierce Chemical Co. ILL) column or Streptavidin-Dynabeads (Dynal Co., Norway) followed by washing and competitive elution as described above. Following each round of bio-panning, the eluted phages were amplified to high titer according to supplier\'s instructions. The amplified fraction was used in the fourth round biopanning. The phage particles from the final fourth round eluate were plated at low density to allow isolation of single phage clones. 1576 CEGFLSAWC CFALFGEEC CFDYGYFWC CFLGAWLGC CGCRGDCVC CGFFLFGEC CGLSLDQWC CGLWAGLFC CGLWIGLWC CHMGFFGEC CLLAAWLGC CLLAGWIPC CLLEAWLGC CMLAGWIPC CMLGEWLGC CPLWAALWC CPLWVGLWC CRGPLRLNC CWALFGESC CWASLFGEC CWGLFGERC CWLAPWLGC CWLSEWLGC 23 Phage display (common panning) 3 PMID:18583538 Human adipose stem cell SPARC Not determined. Not determined. CX7C phage display library Authors demonstrate that both primary and cultured human and mouse stromal cells express a conserved receptor targeted by peptides found to mimic SPARC, a matricellular protein that is required for normal WAT development. A signaling receptor for SPARC has not as yet been determined. By using the SPARC-mimicking peptides CMLAGWIPC and CWLGEWLGC, isolated by panning on human and mouse cells, respectively, we identified the α5β1 integrin complex as a candidate receptor for SPARC. 1577 CTEATVAGC CSRGTVWPC CVGGDSTNC CLMSVGVMC CAHSGVLLC CKGPGSGFC CVDVRSGGC CIVGGVLRC CTRVENGWC CDYSSHGFC CTEHGISGC CLLSVFGAC CRVTLRAAC CARVKSREC CLEAGYSVC CRWGAGRTC CDSSLDWHC CAGHQLIVC CPFLRTLRC CEIALCGPC CSDLVRLGC CWLGEWLGC CETRNVASC CSEWSGSLC CAGALGGSC CFGESRNAC 26 Phage display (common panning) 3 PMID:18583538 Mouse 3T3-L1 cells SPARC Not determined. Not determined. CX7C phage display library Authors demonstrate that both primary and cultured human and mouse stromal cells express a conserved receptor targeted by peptides found to mimic SPARC, a matricellular protein that is required for normal WAT development. A signaling receptor for SPARC has not as yet been determined. By using the SPARC-mimicking peptides CMLAGWIPC and CWLGEWLGC, isolated by panning on human and mouse cells, respectively, we identified the α5β1 integrin complex as a candidate receptor for SPARC. 1578 TPPHRHTHHSTL(1) KPPHSHKHPLLT(1) RYQPHPSKTSTS(1) HIMPHLIPVSVL(1) RTQSQPNRHRPR(1) 5 Phage display (common panning) 9 DOI:10.1021/jp809370z Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Single-walled carbon nanotubes (SWNTs) were grown on Quartz. BSA, TBST 0.1, TBST, and Gly-HCl were used as blocking, incubation, wash, and elution buffer solutions respectively. 1579 APAHLHKPSHVR(1) HGNLHKTHLKLP(1) KQPNTHHVHPHS(1) SPKWHPHHQHWR(1) HLRTHPSHHNVP(1) EDPNLQSSLRMP(1) 6 Phage display (common panning) 9 DOI:10.1021/jp809370z Quartz Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) BSA, TBST 0.1, TBST, and Gly-HCl were used as blocking, incubation, wash, and elution buffer solutions respectively. 1580 HLTPTSTWSNPH(2) APHLQHGHHPHR(1) HPPHHQTHHRTP(1) VPKAHHHLHYEA(1) SLSDYHRSPQLS(1) NPGNYTQYRTTN(1) 6 Phage display (common panning) 9 DOI:10.1021/jp809370z Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Single-walled carbon nanotubes (SWNTs) were grown on Quartz. Blocking buffers were not used in whole experimental procedure. TBST 0.1, water, and Gly-HCl were used as incubation, wash, and elution buffer solutions respectively. 1581 PRPALSTGPGRF(2) RPLYDSYNTGMR(1) RP 2 Phage display (common panning) 9 DOI:10.1021/jp809370z Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Single-walled carbon nanotubes (SWNTs) were grown on SiO2 wafer. Blocking buffers were not used in whole experimental procedure. TBST 0.1, TBST, and Gly-HCl were used as incubation, wash, and elution buffer solutions respectively. 1582 PRPALSTGPGRF(1) PSNKRRKDLANV(1) RP 2 Phage display (common panning) 9 DOI:10.1021/jp809370z SiO2 wafer Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Blocking buffers were not used in whole experimental procedure. TBST 0.1, TBST, and Gly-HCl were used as incubation, wash, and elution buffer solutions respectively. 1583 KPPLHNHHHSLP(6) KPPHHHNHPLTK(2) GPPHYHKHKLSA(1) LPHHGHTHKMRV(1) HKLQHLPPPHLR(4) HPPKKPIMNTML(2) HPKPQHAHLKPV(1) HGTKPPHLHSVR(1) HKQHHSPQNFSL(1) KPPDRHVHKLPI(1) 10 Phage display (common panning) 9 DOI:10.1021/jp809370z Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Single-walled carbon nanotubes (SWNTs) were grown on SiO2 wafer. Blocking buffers were not used in whole experimental procedure. TBS, TBS, and Gly-HCl were used as incubation, wash, and elution buffer solutions respectively. 1584 KPTHLHHHTRIL(3) KPLHVHRHHVMD(1) KPVHPHQHLKLS(1) KPIHHHPHLPLK(1) KPPHSHKHPLLT(1) IPPHPHAHHQKR(1) APKNHVHHVHKS(1) HLGPKHPPKHHH(1) 8 Phage display (common panning) 9 DOI:10.1021/jp809370z SiO2 wafer Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Blocking buffers were not used in whole experimental procedure. TBS, TBS, and Gly-HCl were used as incubation, wash, and elution buffer solutions respectively. 1585 DDWSHWWRAWNG(3) YTSPWWLAWYDP(2) AWWEAFIPNSIT(1) WFPIAWPESWYH(1) GWDWAQDWNWWT(1) NDNPWLMWLKNW(1) YEYPWANWWLSP(1) SSAWWSYWPPVA(6) S-x-W(2)-x(2)-W 8 Phage display (common panning) 9 DOI:10.1021/jp809370z Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Single-walled carbon nanotubes (SWNTs) were grown on SiO2 wafer. Blocking buffers were not used in whole experimental procedure. TBS, TBS, and TBST 0.5 were used as incubation, wash, and elution buffer solutions respectively. 1586 HNKHLPSTQPLA SVSVGMKPSPRP VISNHRESSRPL KSLSRHDHIHHH 4 Phage display (common panning) 5 DOI:10.1021/nl049825n L1(0) phase of FePt, L1(0) FePt Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The sequences contain numerous amines, known to be excellent ligands for Pt, lysine, believed to be essential in the mechanism of the binding of HMG domain proteins to the Pt/DNA complex formed in cisplatin-based cancer therapies, and histidine, which is used extensively to bind Pt salts for studying protein activity and for the heavy metal staining of proteins to facilitate in X-ray crystallography. Basic local alignment search tool (BLAST) searches performed on these peptides showed that they all possess similarities to motifs found in naturally occurring proteins. 1587 YGAFQG(2) YGGFLT(2) YGYWSL(1) YGAFMQ(2) YGAFFQ(1) YGAFFK(2) YGFWSN(1) YGAFGG(1) YGGFGF(1) YGVFSR(1) YGGLSM(2) YGTFLN(2) YGGLVR(1) YGSFSL(2) YGAWYT(2) YGRFFH(1) YGGLRH(1) YGSFMA(1) YGGFSP(1) 19 Phage display (common panning) 2 PMID:1443536 Anti-beta-endorphin monoclonal antibody 3-E7 Fab Beta-endorphin Not determined. Not determined. X6 fAFF1 phage display library Phage (2e9 infectious particles in 100 μ1 of TBS) were added to 100 μ1 of TBS containing biotinylated 3E7 Fab (50 pM final concentration) and incubated overnight at 4°C. The mixture was then exposed to streptavidin-coated plates for 3 h at room temperature and bound phage were isolated after 60 min of washing. 1588 CPTHYISTSLC(3)[2.201] CPTHYTTSAPC(1)[2.554] CPTHYISSRHC(1)[2.369] CPSHYVPRIYC(1)[1.993] CPAHYANYLPC(1)[2.234] CAPSLPAGYLC(1)[NT] CPASSHTILVC(1)[NT] CGKFPGSKPSC(1)[NT] CLPAHGPSLSC(1)[NT] 9 Phage display (common panning) PMID:8666827 Anti-HCV polyclonal antibody Genome polyprotein Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA ELISA measurement of reactivity against the indicated phagotopes of Abs affinity purified from serum C12 using phagotope P621c (displaying CPAHYANYLPC) was done. Average values (OD405 nm to OD620 nm) from two independent experiments were determined, reproduced from the graph and shown. The absorbance of the wild type phage was 0.033. NT denotes not tested. 1589 NKTKQNPNL(1)[0.999] KLRRSTNWG(1)[1.729] KSMRSANKP(1)[1.656] NRRNFTAPV(1)[NT] 4 Phage display (common panning) PMID:8666827 Anti-HCV polyclonal antibody Genome polyprotein Not determined. Not determined. pVIII-9aa and pVIII-9aa.Cys phage display library pool ELISA ELISA measurement of reactivity against the indicated phagotopes of Abs affinity purified from serum C12 using phagotope P787 (displaying KLRRSTNWG) was done. Average values (OD405 nm to OD620 nm) from two independent experiments were determined, reproduced from the graph and shown. The absorbance of the wild type phage was 0.078. NT denotes not tested. 1590 MLPKPSSFPVPG(1) LSSVNSFPVVTP(1) HLQIQPWYPQIS(1) VPWMEPAYQRFL(1) QPWLEQAYYSTF(1) SALLPWPVLVNY(1) ITTPWDEMRSFL(1) HSFLHPWDLFDY(1) 8 Phage display (common panning) 3-5 PMID:11520599 Human neuroblastoma cell line WAC 2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phages associated with WAC 2 cells were recovered in two steps. Cell surface-bound phages were detached by standard acid elution buffer (pH 2.2) for 10 min. Microscopic inspection revealed that this treatment appeared to leave the cells largely intact. In the second step, the cells were collected by centrifugation and lysed by the addition of 1% Triton X-100 in order to recover phages more strongly associated with the cells. Peptides (HLQIQPWYPQIS and VPWMEPAYQRFL) could bind to certain tumor cell types. Thus, these phages and their displayed peptides have the potential to deliver therapeutic agents to the tumors. 1591 GVSKRGLQCHDFISCSGVPW(4) 1 Phage display (common panning) 5 PMID:14500347 CD19+ cells from patient #16 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 4°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1592 NQSIPKVAGDSKVFCWWCAL(9) DMSFQLVTPFLKALPTGWRG(1) 2 Phage display (common panning) 5 PMID:14500347 CD19+ cells from patient #16 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 4°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1593 NQSIPKVAGDSKVFCWWCAL(3) QIMMGPSLGYYMPSESIFAY(1) 2 Phage display (common panning) 5 PMID:14500347 CD19+ cells from patient #16 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 25°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1594 DMSFQLVTPFLKALPTGWRG(4) GGHGRVLWPDGWFSLVGISP(4) QIMMGPSLGYYMPSESIFAY(2) 3 Phage display (common panning) 5 PMID:14500347 CD19+ cells from patient #16 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 25°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1595 NQSIPKVAGDSKVFCWWCAL(9) QSTPPTKHLTIPRHLRNTLI(1) 2 Phage display (common panning) 5 PMID:14500347 CD19+ cells from patient #16 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 37°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1596 NQSIPKVAGDSKVFCWWCAL(1) QIMMGPSLGYYMPSESIFAY(9) 2 Phage display (common panning) 5 PMID:14500347 CD19+ cells from patient #16 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 37°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1597 KPWMTWQWVVEKSSTEGFRT(10) 1 Phage display (common panning) 5 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 4°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1598 GDELGWNWVTIWPKTLSSRV(6) 1 Phage display (common panning) 5 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 4°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1599 MTARPDPMMSTSTTVNTVLI(5) 1 Phage display (common panning) 5 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 37°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1600 LTSPPKEARPSTTVGKSGRE(3) MGTRWQGDGESQHASVGSGS(2) 2 Phage display (common panning) 5 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 37°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1601 KPWMTWQWVVEKSSTEGFRT(2) GDELGWNWVTIWPKTLSSRV(3) 2 Phage display (subtractive panning) 10 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 4°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. Clearing with autologous EBV-immortalized cell line was applied in rounds 6-10. 1602 KPWMTWQWVVEKSSTEGFRT(5) 1 Phage display (subtractive panning) 10 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 4°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. Clearing with autologous EBV-immortalized cell line was applied in rounds 6-10. 1603 ASSWKSFWHPNPVGMTPASS(4) 1 Phage display (subtractive panning) 10 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 37°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. Clearing with autologous EBV-immortalized cell line was applied in rounds 6-10. 1604 LTSPPKEARPSTTVGKSGRE(1) MARTVTANVPGMGEGMVVVP(3) 2 Phage display (subtractive panning) 10 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 37°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. Clearing with autologous EBV-immortalized cell line was applied in rounds 6-10. 1605 LAVTRPSPNKYATVNKSAQA(1) RTSTGPEARDAWMWYWKPPA(3) FGWTWEDSSSNSLYDMSFPH(1) GARSPIRESFGNTVPFDTRF(1) TGQGEREHREGRYTNSEGSD(1) VQTREAGGMYGGPDWWWEWS(1) ARAFSYWSVTDTERVKFFVP(1) LQIGEGVELVMARPIDSDSG(1) 8 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 16; R2, patient 25; R3, patient 42; R4, patient 19; R5, patient 49. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 4°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1606 MGSRSAVGDFESAEGSRRP(3) MARMSTSEVPPVRIATSHSR(1) ISWDIWRWWYTSEDRDAGSA(3) EFGHGSPYETRQHSQARRWL(1) MAQKWGGGQALQGYHIGANA(1) 5 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 16; R2, patient 25; R3, patient 42; R4, patient 19; R5, patient 49. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 25°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1607 VWGMTTSDHQRKTERLDSPE(1) GTFLHSRSQTSAEERAGGLG(1) GWWQFWDWSARRQDRGEALL(4) MAMGGKPERPADSDNVQVRG(1) QTIWRGHAASVNDSTTVSRT(1) MTSAQTSEKLKAETDRHTAE(1) 6 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 16; R2, patient 25; R3, patient 42; R4, patient 19; R5, patient 49. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 37°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1608 GAGNHPDAVSYPDILVKPRL(4) TAYTTGIRWSSQLQRLQHAS(1) 2 Phage display (subtractive panning) 4 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-4. The cell were incubated for 1 h with the phage at 4°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1609 GAGNHPDAVSYPDILVKPRL(3) TAYTTGIRWSSQLQRLQHAS(1) TDAMLGATVREHLEAMALVG(1) 3 Phage display (subtractive panning) 4 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-4. The cell were incubated for 1 h with the phage at 4°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1610 GAGNHPDAVSYPDILVKPRL(2) TAYTTGIRWSSQLQRLQHAS(1) MPHLDRVLWKPSGIRDLVSE(1) 3 Phage display (subtractive panning) 4 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-4. The cell were incubated for 1 h with the phage at 37°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1611 GAGNHPDAVSYPDILVKPRL(1) TAYTTGIRWSSQLQRLQHAS(2) VTGAVWMVLEESSHVLL(1) EYRVENRIQPIRAYSGSLNR(1) 4 Phage display (subtractive panning) 4 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-4. The cell were incubated for 1 h with the phage at 37°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1612 KPWMTWQWVVEKSSTEGFRT(6) GAGNHPDAVSYPDILVKPRL(1) TQGRMAFYQTSVVLSSADST(1) 3 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 38; R5, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 4°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1613 TAYTTGIRWSSQLQRLQHAS(1) 1 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 38; R5, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 4°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1614 GAGNHPDAVSYPDILVKPRL(2) TAYTTGIRWSSQLQRLQHAS(1) TQGRMAFYQTSVVLSSADST(1) WGTKPTTQWRKPQLQEEVRP(3) 4 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 38; R5, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 37°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1615 TAYTTGIRWSSQLQRLQHAS(2) WGTKPTTQWRKPQLQEEVRP(3) MGRTVQSGDGTPAQTQPSVN(3) 3 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 38; R5, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 37°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1616 RGDKSRPPVWYVEGE(7)[1.085 ± 0.011, 37.181 ± 5.147, 1.069 ± 0.051, 0.026 ± 0.005] VQRGAWRWPVVTNE(4)[0.242 ± 0.017, 10.291 ± 1.133, 1.083 ± 0.031, 0.012] PAGRQARRPATVPEG(1)[0.074, 12.289, 1.013 ± 0.016, 0.025] PEWWLEEPPFISSGK(1)[0.199 ± 0.014, 4.433 ± 0.346, 1.083 ± 0.041, 0.005 ± 0.002] 4 Phage display (common panning) 2 PMID:20708963 Anti-CPS monoclonal antibody 13D9 Capsular polysaccharide, CPS Not determined. Not determined. X15 PC89 phage display library ELISA The affinities of selected phages to anti-CPS monoclonal antibody 13D9 were measured by phage ELISA, phage UltramicroELISA (UMELISA), phage sandwich ELISA and inhibition ELISA. In phage ELISA, the absorbance at 492 nm was determined. The absorbance of the control phage PC89 was 0.086 ± 0.011. In UMELISA, fluorescence units (FU) were read in a photometer-fluorimeter plate reader PR-521. The fluorescence units of the control phage PC89 were 2.436. In sandwich ELISA, the absorbance at 492 nm was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.087 ± 0.020 and 0.077 ± 0.020, respectively. In inhibition ELISA, the absorbance at 492 was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.119 ± 0.008 and 0.188 ± 0.006, respectively. All data were expressed as means ± SD and were reproduced from the graph. Immunization of mice with the phage-displayed peptides elicited anti-peptide IgG antibodies. 1617 RGDKSRPPVWYVEGE(20)[0.380 ± 0.033, 1.069 ± 0.051, 0.026 ± 0.005] 1 Phage display (common panning) 3 PMID:20708963 Anti-CPS monoclonal antibody 13D9 Capsular polysaccharide, CPS Not determined. Not determined. X15 PC89 phage display library ELISA The affinities of selected phages to anti-CPS monoclonal antibody 13D9 were measured by phage ELISA, phage UltramicroELISA (UMELISA), phage sandwich ELISA and inhibition ELISA. In phage ELISA, the absorbance at 492 nm was determined. The absorbance of the control phage PC89 was 0.086 ± 0.011. In UMELISA, fluorescence units (FU) were read in a photometer-fluorimeter plate reader PR-521. The fluorescence units of the control phage PC89 were 2.436. In sandwich ELISA, the absorbance at 492 nm was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.087 ± 0.020 and 0.077 ± 0.020, respectively. In inhibition ELISA, the absorbance at 492 was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.119 ± 0.008 and 0.188 ± 0.006, respectively. All data were expressed as means ± SD and were reproduced from the graph. Immunization of mice with the phage-displayed peptides elicited anti-peptide IgG antibodies. 1618 RGDKSRPPVWYVEGE(25)[0.380 ± 0.033, 1.069 ± 0.051, 0.026 ± 0.005] 1 Phage display (common panning) 4 PMID:20708963 Anti-CPS monoclonal antibody 13D9 Capsular polysaccharide, CPS Not determined. Not determined. X15 PC89 phage display library ELISA The affinities of selected phages to anti-CPS monoclonal antibody 13D9 were measured by phage ELISA, phage UltramicroELISA (UMELISA), phage sandwich ELISA and inhibition ELISA. In phage ELISA, the absorbance at 492 nm was determined. The absorbance of the control phage PC89 was 0.086 ± 0.011. In UMELISA, fluorescence units (FU) were read in a photometer-fluorimeter plate reader PR-521. The fluorescence units of the control phage PC89 were 2.436. In sandwich ELISA, the absorbance at 492 nm was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.087 ± 0.020 and 0.077 ± 0.020, respectively. In inhibition ELISA, the absorbance at 492 was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.119 ± 0.008 and 0.188 ± 0.006, respectively. All data were expressed as means ± SD and were reproduced from the graph. Immunization of mice with the phage-displayed peptides elicited anti-peptide IgG antibodies. 1619 ATNAEPS(1) GGKRLPR(1) GRKLKPR(1) GTGKRPR(1) KVSRPVR(1) LDEEPPR(1) LGGKKKR(1) NKVRLPR(1) PSRRKLR(1) REGRVRR(1) RGGRGPR(1) RGGRPAR(1) RKGRPSR(1) RVMRGPR(1) RWEKVPR(1) TVKKNKR(1) VLTRGLR(1) VMHKPAR(1) VMVRPPR(1) VRSKAPR(1) VSERRPR(1) [RK]-x(2)-R 21 Phage display (common panning) 3 PMID:21486998 Neuropilin-1 Not determined. Not determined. Not determined. X7 T7 phage display library PPC-1 cells were selected as a model because they express high levels of NRP-1. Selection was performed against PPC-1 cells by the microfluidic phage selection (MiPS) platform. The ability of this system to directly target membrane-bound proteins on cell surfaces is significant because most of these proteins are notoriously difficult to produce using recombinant technology. Authors show that this system offers significant advantages compared to conventional biopanning methods. Authors found that the MiPS system enables more efficient enrichment of phage displaying this higher-affinity motif: 90% of the phage selected using the MiPS contained (R/K)XX(R/K) motifs, while 5% displayed lower affinity XXXR motifs. 1620 AQVRGPR(2) RTPR(1) STVRRRR(1) MKIRGPR(1) GRVRGVR(1) LKIERSPR(1) RRKR(1) GKPRAKR(1) AMKRVAR(1) QRKGRR(1) VRKFVIR(1) TNRVRRR(1) RGSQRAR(1) KRLNSGR(1) KMRSSLR(1) IGQDSHR(1) NETRSGN(1) RKRRSVL(1) EVEGCEL(1) VGKKLP(1) [RK]-x(2)-[RK] 20 Phage display (common panning) 3 PMID:21486998 Neuropilin-1 Not determined. Not determined. Not determined. X7 T7 phage display library PPC-1 cells were selected as a model because they express high levels of NRP-1. Authors found that conventional cell suspension-based panning yielded at best 52% (R/K)XX(R/K) phage and 29% XXXR phage. 1621 RIGRPLR(1)[5567, 0.78] GGKRPAR(1)[4645, 1.51] FNRKERR(1)[2650, NT] GKGRAQR(1)[1045, NT] SRAKKPR(1)[912, NT] NGARKPR(1)[817, 3.29] LRKRAAR(1)[581, NT] KKVRVVR(1)[474, NT] GKVGSRR(1)[377, NT] VYKARTR(1)[293, 10.9] LSSAQDR(1)[14, NT] VGSGKRS(1)[1.4, NT] SMTARSV(1)[0.8, NT] GGRGARA(1)[0.6, NT] RVRAGHH(1)[0.8, NT] [RK]-x(2)-R 15 Phage display (common panning) 3 PMID:21486998 Neuropilin-1 Not determined. Not determined. Not determined. X7 T7 phage display library Competition experiment Fifteen phage clones were randomly picked from the R3 pool and their binding to PPC-1 relative to M21 melanoma cells was measured. Of note, in contrast to PPC-1 cells, M21 cells express only minimal amounts of NRP-1 protein. The binding fold was shown. Besides, the equilibrium dissociation constants (Kd, μM) of the peptide sequences for binding to the NRP-1 protein were measured and shown. Three rounds of selection in MiPS device revealed peptides P7 (RIGRPLR) and P4 (GGKRPAR) exhibiting higher affinity and specificity in binding to cells expressing NRP-1 compared with the well known CR (RPARPAR) with a Kd of 2.79 μM. NT denotes not tested. PPC-1 cells were selected as a model because they express high levels of NRP-1. Selection was performed against PPC-1 cells by the microfluidic phage selection (MiPS) platform. The ability of this system to directly target membrane-bound proteins on cell surfaces is significant because most of these proteins are notoriously difficult to produce using recombinant technology. Authors show that this system offers significant advantages compared to conventional biopanning methods. 1622 AKAFSYWTPFDGKSLYSST(1)[0.581 ± 0.016] IVTARARHDWNWVQRSSRSL(1)[0.047] TYNHDYKTGMVIYSPFMTYP(1)[1.791 ± 0.010] 3 Phage display (common panning) 3 PMID:21697352 Anti-NSP2 monoclonal antibody Non-structural protein 2, NSP2 Not determined. Not determined. X20 fUSE5 phage display library ELISA In phage ELISA, the absorbance at 490 nm was determined. Data shown were reproduced from the graph and expressed as means ± SD. BLAST and ClustalW2 analysis of the phage clone TYNHDYKTGMVIYSPFMTYP peptide and NSP2 protein sequences located a motif (244)T-(Y/F)-Φ-Φ-Φ-X-K-Φ-G(252), in the C-terminal domain of the protein, where Φ is a hydrophilic residue and amino acids at position X249 did not display conserved/similar properties. This motif was found to be conserved from analyzing an alignment of Australian and GenBank human NSP2 sequences, which also suggests that antibodies against NSP2 may provide heterotypic protection. 1623 TYNHDYKTGMVIYSPFMTYP 1 Phage display (common panning) 4 PMID:21697352 Anti-NSP2 monoclonal antibody Non-structural protein 2, NSP2 Not determined. Not determined. X20 fUSE5 phage display library BLAST and ClustalW2 analysis of the phage clone TYNHDYKTGMVIYSPFMTYP peptide and NSP2 protein sequences located a motif (244)T-(Y/F)-Φ-Φ-Φ-X-K-Φ-G(252), in the C-terminal domain of the protein, where Φ is a hydrophilic residue and amino acids at position X249 did not display conserved/similar properties. This motif was found to be conserved from analyzing an alignment of Australian and GenBank human NSP2 sequences, which also suggests that antibodies against NSP2 may provide heterotypic protection. 1624 TYNHDYKTGMVIYSPFMTYP 1 Phage display (common panning) 5 PMID:21697352 Anti-NSP2 monoclonal antibody Non-structural protein 2, NSP2 Not determined. Not determined. X20 fUSE5 phage display library BLAST and ClustalW2 analysis of the phage clone TYNHDYKTGMVIYSPFMTYP peptide and NSP2 protein sequences located a motif (244)T-(Y/F)-Φ-Φ-Φ-X-K-Φ-G(252), in the C-terminal domain of the protein, where Φ is a hydrophilic residue and amino acids at position X249 did not display conserved/similar properties. This motif was found to be conserved from analyzing an alignment of Australian and GenBank human NSP2 sequences, which also suggests that antibodies against NSP2 may provide heterotypic protection. 1625 TYNHDYKTGMVIYSPFMTYP 1 Phage display (common panning) 6 PMID:21697352 Anti-NSP2 monoclonal antibody Non-structural protein 2, NSP2 Not determined. Not determined. X20 fUSE5 phage display library BLAST and ClustalW2 analysis of the phage clone TYNHDYKTGMVIYSPFMTYP peptide and NSP2 protein sequences located a motif (244)T-(Y/F)-Φ-Φ-Φ-X-K-Φ-G(252), in the C-terminal domain of the protein, where Φ is a hydrophilic residue and amino acids at position X249 did not display conserved/similar properties. This motif was found to be conserved from analyzing an alignment of Australian and GenBank human NSP2 sequences, which also suggests that antibodies against NSP2 may provide heterotypic protection. 1626 APLLTWPRAIGP(0.166) MNNSLLPMRLQT(0.166) VPVQTQRNFLSI(0.166) DIHSPTRQSPYY(0.166) SPSHQWPSPKGS(0.166) SSSQTDNRMSAG(0.166) 6 Phage display (common panning) 3 PMID:19539948 Semiconductor InP(001) surface Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1627 HAPGNMLRVSLG(0.5) YAIKGPSHFRPS(0.5) 2 Phage display (common panning) 4 PMID:19539948 Semiconductor InP(001) surface Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1628 SVSVGMKPSPRP(1.0) 1 Phage display (common panning) 6 PMID:19539948 Semiconductor InP(001) surface Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1629 AANDKMQKFRLV(0.111) GFDKIPLDMMRG(0.111) ITPHHATLPQSR(0.111) DYQFKSSPLRET(0.111) KWPGNSMFPYGF(0.111) QYPYLLSAGDIH(0.111) QSTPTRPGMLLL(0.111) TNKPSFTPLTAS(0.111) YTITPNPLPSNP(0.111) 9 Phage display (common panning) 3 PMID:19539948 Semiconductor InP(111) surface Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1630 SVSVGMKPSPRP(0.8) SSPLHQQSHYTY(0.2) 9 Phage display (common panning) 4 PMID:19539948 Semiconductor InP(111) surface Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1631 SVSVGMKPSPRP(1.0) 9 Phage display (common panning) 6 PMID:19539948 Semiconductor InP(111) surface Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1632 CIFPLCLGRPIAASYPPKTR(1)[0.908 ± 0.041, 0.598 ± 0.008] LNSRHTDKGTVQFALIAVRG(1)[1.064 ± 0.065, 0.957 ± 0.010] YVALQGSMFDRVRVFWMARG(1)[0.826 ± 0.033, 0.442 ± 0.017] ACSDRFLRRPMCLPPHVVFL(1)[1.146 ± 0.008, 0.303 ± 0.025] GFGKDITTGHSFTHKSNNDP(1)[0.836 ± 0.029, 0.373 ± 0.024] TTGNLSLDLLIATRFSSHGK(1)[0.948 ± 0.021, 0.880] IHTDRPAWKYVGALPVIHVR(1)[0.220 ± 0.006, 0.223 ± 0.024] NSNFTVHSNYWYWNRYIPPV(1)[1.046 ± 0.019, 0.945 ± 0.009] TNLLYMLNTEAQYKALFMRK(1)[0.871 ± 0.024, 0.492] ILPNPIHHTWLPFKLSHNLP(1)[0.609 ± 0.019, 0.557 ± 0.062] GCSYPMCRFNVVRFTGLSLL(1)[1.166 ± 0.009, 0.680 ± 0.021] GLFDLTNYYYLSRGTQIKGT(1)[0.707, 0.663 ± 0.022] 12 Phage display (common panning) 3 PMID:19095030 Anti-EG95 polyclonal antibody EG95 Not determined. Not determined. X20 fUSE5 phage display library ELISA Phage binding to anti-EG95 polyclonal antibody was measured in the absence or presence of the protein competitor (EG95-MBP) by ELISA. The absorbance at 450 nm was determined. Displayed values are reproduced from the graph and expressed as the mean ± SEM of duplicate wells. The absorbance of the phage R1 as negative control was 0.110 both in the absence or presence of the protein competitor (EG95-MBP). 1633 NRLFDRVDRFYA(1)[1.292 ± 0.065, 0.975 ± 0.057] GKPGWLLDNVAL(1)[0.851 ± 0.022, 0.108 ± 0.014] HYKWLNDPLAAR(1)[0.916 ± 0.036, 0.426 ± 0.007] TLFGRMEHYFN(1)[1.277 ± 0.054, 1.123 ± 0.023] ISTSKPAWKLAN(1)[1.268 ± 0.050, 0.186 ± 0.032] CIPPLCKLRLHE(1)[1.168 ± 0.100, 0.541 ± 0.087] LDKRNNDPFHLP(1)[0.904 ± 0.168, 0.123 ± 0.018] SVSVGMKPSPRP(1)[0.474 ± 0.014, 0.371 ± 0.015] 8 Phage display (common panning) 4 PMID:19095030 Anti-EG95 polyclonal antibody EG95 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Phage binding to anti-EG95 polyclonal antibody was measured in the absence or presence of the protein competitor (EG95-MBP) by ELISA. The absorbance at 450 nm was determined. Displayed values are reproduced from the graph and expressed as the mean ± SEM of duplicate wells. The absorbance of the phage R1 as negative control was 0.110 both in the absence or presence of the protein competitor (EG95-MBP). 1634 VSQTMRQTAVPLLWFWTGSL YAAWPASGAWTGTAPCSAGT 2 Phage display (common panning) 5 PMID:16386888 H1299 cell line Not determined. Not determined. Not determined. IR20 phage display library (X20) A cell-targeting Peptide YAAWPASGAWTGTAPCSAGT, which represented 10 of the 12 clones sequenced, was isolated at round 5. The isolated phage clone binds H1299 cells 80 times better than a control phage and can distinguish between H1299 and normal control cells. The phage clone also binds to the lung pleura epidermoid cell line, Calu-1 but not to all lung carcinoma cell lines. The peptide is functional outside the context of the phage and tetramerization of the peptide on a trilysine core improves the affinity of the peptide. 1635 SHRLHAT*ILMP(1)[1.680] SHRILSSALV*V(1)[1.530] SHRLHNTMPSES(1)[2.006] THRLYLASTLPG(1)[1.685] SHRLPHPNLIRL(1)[1.830] QHRIHNPAPVSL(1)[2.083] HHRILIEPPSMH(1)[1.685] SHRLPPVSNPSL(1)[1.607] HHRLNNQDDLLK(1)[1.860] SHKLFYQMGTQL(1)[1.573] QHRMTWMQGLSS(1)[1.685] QHKLYPVIPLPS(1)[2.100] H-[RK]-[LI] 12 Phage display (common panning) 3 PMID:13679612 Anti-DENV2 monoclonal antibody Ab4 Dengue virus 2, DENV2 Not determined. Not determined. X12 phage display library ELISA In phage ELISA, the absorbance at 490 nm was measured. Data shown were reproduced from the graph. Selected phage clones reacted specifically with Ab4 and did not react with other mAbs. Immunopositive phage clones displayed a consensus motif, His-Arg/Lys-Leu/Ile, and a synthetic peptide corresponding to the phage-displayed peptide bound specifically to Ab4. The His and Arg residues in this epitope were found to be crucial for peptide binding to Ab4 and binding activity decreased dramatically when these residues were changed to Leu. The epitope-based synthetic peptide not only identified serum samples from DEN-2-immunized mice and rabbits by ELISA but also differentiated clearly between serum samples from DEN-2- and Japanese encephalitis virus-immunized mice. 1636 SMSIARL VSFLEYR 2 Phage display (in vivo) PMID:11830668 Mouse prostate Not determined. Not determined. Not determined. X12 phage display library For the library screening, CD-1 mice were anesthetized with Avertin (0.015 ml/g) and injected intravenously (tail vein) with phage libraries containing e9 transducing units diluted in 200 μl of DMEM. The phage was rescued from tissues by bacterial infection, and about 300 individual colonies were grown separately. The phage bound also to vasculature in the human prostate. The peptide displayed by the prostate-homing phage, SMSIARL, was synthesized and shown to inhibit the homing of the phage when co-injected into mice with the phage. 1637 CNLYSCLK AREYRWLG 2 Phage display (common panning) 5 PMID:11562257 Anti-phosphotyrosine monoclonal antibody PY20 Phosphotyrosine Not determined. Not determined. pGWX3YX4 phage display library Western blot Among the randomly picked clones from pGWX3YX4, a band at 24.5 kD by Western blot, that corresponded to the gIII product containing the displayed peptide, was present in 2 clones CNLYSCLK and AREYRWLG, and it was absent from the control phage derived from the parent pGWg3 vector. This was proof that the displayed peptide was a Tie-2 substrate in the phage context. The phage libraries were phosphorylated using a fusion protein of GST and human Tie-2 kinase domain (hGST-Tie-2). 1638 EAIVYVWGR RLVAYEGWV 2 Phage display (common panning) 5 PMID:11562257 Anti-phosphotyrosine monoclonal antibody PY20 Phosphotyrosine Not determined. Not determined. pGWX4YX4 phage display library ELISA Anti-phosphotyrosine ELISA of clones from pGWX4YX4 also identified 2 positive clones EAIVYVWGR and RLVAYEGWV with a signal over background >2 (data not shown). The phage libraries were phosphorylated using a fusion protein of GST and human Tie-2 kinase domain (hGST-Tie-2). 1639 LPLTPLP 1 Phage display (common panning) 3 PMID:17360703 Peptide IDQYRMQRDKEFRLKQ Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The most common phage sequence observed was LPLTPLP. When this sequence was used to search the human genome data base using the \"short, nearly exact match\" BLAST algorithm, an exact match of phage peptide amino acids 2-7 was found to amino acids 85-90 of the H(+)-ATPase subunit a4. This sequence is PLTPLP in a4 and PEVPFP in a1. 1640 WHWQRPLMPVSI(0.7) WHWSPRTALYTT(0.05) WHWNFKPPHDLL(0.06) WHWKPPAPYVWW(0.12) 4 Phage display (common panning) 4 PMID:18363413 Trinitrotoluene, TNT Not determined. Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool 1641 HPNFSKYILHQR(0.24) QRPTTQQGPSML(0.24) QRPTTQLGSEYA(0.06) 3 Phage display (common panning) 5 PMID:18363413 2,4-dinitrotoluene, DNT Not determined. Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool 1642 HVGGSSV SLRGDGSSV SVRGSGSGV SVGSRV SVVRDGSEV SGRKVGSGSSV SRKQGGTEV 7 Phage display (in vivo) 6 PMID:18297085 Human lewis lung carcinoma, LLC Not determined. Not determined. Not determined. T7 phage display library 1643 HVGGSSV SLRGDGSSV SVGSRV SVVRDGSEV SGRKVGSGSSV SRKQGGTEV 6 Phage display (in vivo) 6 PMID:18297085 Mouse GL261 glioblastoma Not determined. Not determined. Not determined. T7 phage display library 1644 MSGDIISLAPTG GPFFPKSLTTTS NAPLSHIPENPR SISAMPAPANSS SVSVGMKPSPRP 5 Phage display (common panning) 3 PMID:18582017 Semiconductor GaN(0001) surface Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) As the abundance of the last sequence was 60%, hence revealing a high affinity for the test surface, the sequence SVSVGMKPSPRP was selected as the specific peptide for the GaN (0001) surface, denoted "GaN_probe" in the following. This sequence presents a hydrophobic first half and a hydrophilic second half. 1645 TDSILRSYDWTY(1) TQQPLEGHQLPY(1) TGVSWSVAQPSF(1) SVSVGMKPSPRP(1) SQWNSPPSSAAF(1) 5 Phage display (common panning) 3 PMID:19137069 Non-small cell lung cancer (CL1-5) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The targeting phage TDSILRSYDWTY bound to several NSCLC cell lines but not to normal cells. Both the targeting phage and the synthetic peptide recognized the surgical specimens of NSCLC with a positive rate of 75% (27 of 36 specimens). In severe combined immunodeficiency (SCID) mice bearing NSCLC xenografts, the targeting phage specifically bound to tumor masses. The tumor homing ability of the targeting phage was inhibited by the cognate synthetic peptide, but not by a control or a WTY-mutated peptide. 1646 TDSILRSYDWTY(20) DMPKQLLAPWYY(2) SYPLSFLGPLIS(1) W-[TY]-Y 3 Phage display (common panning) 5 PMID:19137069 Non-small cell lung cancer (CL1-5) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The targeting phage TDSILRSYDWTY bound to several NSCLC cell lines but not to normal cells. Both the targeting phage and the synthetic peptide recognized the surgical specimens of NSCLC with a positive rate of 75% (27 of 36 specimens). In severe combined immunodeficiency (SCID) mice bearing NSCLC xenografts, the targeting phage specifically bound to tumor masses. The tumor homing ability of the targeting phage was inhibited by the cognate synthetic peptide, but not by a control or a WTY-mutated peptide. 1647 TDSILRSYDWTY(27) DMPKQLLAPWYY(3) W-[TY]-Y 2 Phage display (common panning) 5 PMID:19137069 Non-small cell lung cancer (CL1-5) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The targeting phage TDSILRSYDWTY bound to several NSCLC cell lines but not to normal cells. Both the targeting phage and the synthetic peptide recognized the surgical specimens of NSCLC with a positive rate of 75% (27 of 36 specimens). In severe combined immunodeficiency (SCID) mice bearing NSCLC xenografts, the targeting phage specifically bound to tumor masses. The tumor homing ability of the targeting phage was inhibited by the cognate synthetic peptide, but not by a control or a WTY-mutated peptide. 1648 CMLPHHGAC CNPGFAQAC 2 Phage display (common panning) 1-3 PMID:18271563 Hydroxyapatite, HA Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) After three biopanning rounds, 56 clones were selected (18 from first round, 19 each from second and third rounds) and characterized in terms of their physicochemical properties and binding affinity to HA. On the basis of immunofiuorescence microscopy assays, out of 56 clones, 12 were characterized as strong binders with a surface coverage higher than 40%, 28 were moderate binders with a coverage between 35% and 10%, and 16 were weak binders with a coverage lower than 7%. In addition, ELISA was used to confirm these results and the data obtained from both techniques were consistent. Statistical analysis of the binders after binding characterization revealed that 67% of the strong binders were selected in the third round of panning; likewise, 69% of the weak binders were selected in the first round of panning. These results showed that selection of strong-binding clones was favored in the additional panning rounds.The strongest binding clone (CMLPHHGAC, 75 + 5% binding) and a weak-binding clone (CNPGFAQAC, 3 + 2% binding) were chosen for subsequent solid-phase peptide synthesis and mineralization experiments. 1649 DRHSRIVILMPLAYP 1 Phage display (common panning) 4 PMID:11707616 Anti-AMA-1 monoclonal antibody 5G8 Apical membrane antigen 1 Not determined. Not determined. f3-15mer phage display library (X15) ELISA Selected clones after four rounds of panning were shown to bind to MAb5G8-coated ELISA plates, in a dose-dependent manner, whereas a clone containing a peptide isolated by panning the same library on an unrelated protein was unable to bind MAb5G8 even at 10e11 CFU/ml. 1650 SWGPFPF(4) SWGPLPF(1) 2 Phage display (common panning) 4 PMID:10427007 Anti-DON monoclonal antibody 6F5 Mycotoxin deoxynivalenol, DON Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The absorbance was measured at 450 nm (data not shown). In ELISA, five of phage isolates exhibited binding to mAb 6F5. In competitive ELISA, binding of phage clones SWGPFPF and SWGPLPF to immobilized mAB 6F5 was competitively inhibited by free DON. Fifteen individual phage plaques from the fourth-round-selected phages were randomly isolated and used to infect E. coli ER2537 for phage reamplification. Each of the 15 phage isolates was tested to determine its binding to mAb 6F5 by ELISA. Five of these phage isolates exhibited binding to mAb 6F5. 1651 CGSYWHPQC(2) CGTFIHPQC(3) HPQ 2 Phage display (common panning) 3 PMID:16258189 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. Ph.D.-C7C phage display library (CX7C) 1652 SLMVSVFPFLWMDLVW(7) RRFPLYFLSELMELIL(5) SGTWAVFPFLWLDLMW(4) SLMMSVFPFLWMDLVW(1) SSMVSVFPFLWMDLVW(1) SLMVSVFPFLWMDLMW(1) CVSWLGFAWEWLQLLG(1) GSSFWAHWEKLLSESL(1) FAWHWKILSKFCLWLG(1) PRFPLYFLSELMELIL(1) RFYFLSALVLVLSGKL(1) LQVALFLVGLTRGLSE(1) VGFSDARALLFIVLEL(1) RLFIRARCKLLLLLRL(1) FLRLLARLFISCSVVI(1) FLCLLSLLHRSYFFGI(1) LCLIVCRRRLLVRVCF(1) LCICVVVRVLLWSSVQ(1) SMPFLFHWLNLYLHVC(1) GSAGYLWLLVMRSICV(1) F-x(3)-W-x(2)-L 20 mRNA display (common panning) 4 PMID:21423613 E3 ubiquitin-protein ligase Mdm2 Cellular tumor antigen p53 1YCR,4HFZ, Not determined. X16 mRNA display library The mRNA-displayed peptide library is incubated with MDM2 immobilized on beads through an affinity selection tag containing a ZZ domain and a TEV protease cleavage site, and unbound molecules are washed away. The bound molecules are eluted by cleavage with the TEV protease, and their mRNA portion is amplified by RT-PCR. The resulting DNA can be used for the next rounds of selection or analyzed by cloning and sequencing. While previous studies have used phage display to identify peptides (such as DI) that inhibit the MDM2-p53 interaction, these peptides were not sufficiently optimized because the size of the phage-displayed random peptide libraries did not cover all of the possible sequences. In this study, authors performed selection of MDM2-binding peptides from large random peptide libraries in two stages using mRNA display. Authors identified an optimal peptide (PRFWEYWLRLME) named MIP that inhibited the MDM2-p53 and MDMX-p53 interactions 29- and 13-fold more effectively than DI, respectively. Expression of MIP fused to the thioredoxin scaffold protein in living cells by adenovirus caused stabilization of p53 through its interaction with MDM2, resulting in activation of the p53 pathway. Furthermore, expression of MIP also inhibited tumor cell proliferation in a p53-dependent manner more potently than DI. 1653 PRFWEYWLRLME(10) KSFQQYWQELML(9) KTFEEYWLMLMS(4) PSFWEHWVELML(4) KRFQDYWSELML(3) 5 mRNA display (common panning) 5 PMID:21423613 E3 ubiquitin-protein ligase Mdm2 Cellular tumor antigen p53 1YCR,4HFZ, Not determined. X(2)-F-X(3)-W-X(2)-L-X(2) mRNA display library ELISA In ELISA, MIP (PRFWEYWLRLME) could inhibit the MDM2-p53 interaction with an IC50 of 10 nM. Also MIP could inhibit the MDMX-p53 interaction at an IC50 of 120 nM. The mRNA-displayed peptide library is incubated with MDM2 immobilized on beads through an affinity selection tag containing a ZZ domain and a TEV protease cleavage site, and unbound molecules are washed away. The bound molecules are eluted by cleavage with the TEV protease, and their mRNA portion is amplified by RT-PCR. The resulting DNA can be used for the next rounds of selection or analyzed by cloning and sequencing. While previous studies have used phage display to identify peptides (such as DI) that inhibit the MDM2-p53 interaction, these peptides were not sufficiently optimized because the size of the phage-displayed random peptide libraries did not cover all of the possible sequences. In this study, authors performed selection of MDM2-binding peptides from large random peptide libraries in two stages using mRNA display. Authors identified an optimal peptide (PRFWEYWLRLME) named MIP that inhibited the MDM2-p53 and MDMX-p53 interactions 29- and 13-fold more effectively than DI, respectively. Expression of MIP fused to the thioredoxin scaffold protein in living cells by adenovirus caused stabilization of p53 through its interaction with MDM2, resulting in activation of the p53 pathway. Furthermore, expression of MIP also inhibited tumor cell proliferation in a p53-dependent manner more potently than DI. 1654 INTLLSEINSILLDIISLL(13)[2.709 ± 0.168] INLLLMEINMILNEIITLL(3)[3.409 ± 0.061] INTLLMEINNILLNIIDFL(2)[3.088 ± 0.161] INTLLEEINIILNNIISFL(2)[3.079 ± 0.124] INSLLKEINNILISIIDFL(2)[2.709 ± 0.113] INSLLMEINTILNDIIMFL(1)[2.983 ± 0.185] ISSLLEEINTILMDIIEYL(1)[2.726 ± 0.079] 7 mRNA display (competitive panning) 10 PMID:21136209 Interleukin-6, IL-6 Interleukin-6 receptor 1P9M, Not determined. X(3)-L-I-X-E-I-X(2)-I-L-X(2)-I-X(3)-L mRNA display library ELISA In peptide ELISA, the absorbance at 490 nm was determined. Data were reproduced from the graph and expressed as the mean ± SD of triplicate determinations. The phage RNA(-) acted as negative control with A490 of 0.037. Beside, both RA07 (INTLLSEINSILLDIISLL) and CA11 (NQQLIEEIIQILHKIFEIL) peptides bound to GST-IL-6 in a dose-dependent manner and RA07 had higher affinity for GST-IL-6 (EC50=45.1 nM) than CA11 (EC50=142 nM). To determine whether CA11 and RA07 have inhibitory activity against IL-6, a KT-3 cell-based assay was performed. KT-3 cells are known to proliferate in response to IL-6. Results showed that RA07 suppressed KT-3 cell proliferation in a dose-dependent manner, but CA11 did not. The IC50 value of RA07 was 5.6 μM. Besides, the results of ELISA-based IL-6 inhibition assay suggested that RA07 inhibits gp130 from binding to the IL-6/IL-6R complex, but not IL-6R from binding to IL-6. The IC50 value of RA07 in this assay was 2.8 μM, which was almost the same as in the KT-3 cell-based assay (5.6 μM). After the beads were washed three times with selection buffer, 1-8 μM of IL-6 (Ajinomoto) was added and incubation continued another 2 h. The bound molecules were eluted with PreScission protease (GE healthcare). The most frequently occurring sequence, RA07 (INTLLSEINSILLDIISLL), bound to IL-6 with 3 to 4-fold higher affinity than CA11. Furthermore, RA07 inhibited IL-6-dependent KT-3 cell proliferation in a dose-dependent manner. ELISAs revealed that RA07 could not inhibit IL-6 from binding to the IL-6 receptor (IL-6R), but could inhibit the IL-6/IL-6 complex binding to gp130. 1655 INTLLSEINSILLDIISLL(6)[2.709 ± 0.168] INTLLMEINNILLNIIDFL(2)[3.088 ± 0.161] INSLLMEINTILNDIIMFL(2)[2.983 ± 0.185] ISSLLEEINTILMDIIEYL(2)[2.726 ± 0.079] INSLLKEINNILISIIDFL(1)[2.709 ± 0.113] 5 mRNA display (competitive panning) 10 PMID:21136209 Interleukin-6, IL-6 Interleukin-6 receptor 1P9M, Not determined. X(3)-L-I-X-E-I-X(2)-I-L-X(2)-I-X(3)-L mRNA display library ELISA In peptide ELISA, the absorbance at 490 nm was determined. Data were reproduced from the graph and expressed as the mean ± SD of triplicate determinations. The phage RNA(-) acted as negative control with A490 of 0.037. Beside, both RA07 (INTLLSEINSILLDIISLL) and CA11 (NQQLIEEIIQILHKIFEIL) peptides bound to GST-IL-6 in a dose-dependent manner and RA07 had higher affinity for GST-IL-6 (EC50=45.1 nM) than CA11 (EC50=142 nM). To determine whether CA11 and RA07 have inhibitory activity against IL-6, a KT-3 cell-based assay was performed. KT-3 cells are known to proliferate in response to IL-6. Results showed that RA07 suppressed KT-3 cell proliferation in a dose-dependent manner, but CA11 did not. The IC50 value of RA07 was 5.6 μM. Besides, the results of ELISA-based IL-6 inhibition assay suggested that RA07 inhibits gp130 from binding to the IL-6/IL-6R complex, but not IL-6R from binding to IL-6. The IC50 value of RA07 in this assay was 2.8 μM, which was almost the same as in the KT-3 cell-based assay (5.6 μM). The bound molecules were eluted with PreScission protease (GE healthcare). The most frequently occurring sequence, RA07 (INTLLSEINSILLDIISLL), bound to IL-6 with 3 to 4-fold higher affinity than CA11. Furthermore, RA07 inhibited IL-6-dependent KT-3 cell proliferation in a dose-dependent manner. ELISAs revealed that RA07 could not inhibit IL-6 from binding to the IL-6 receptor (IL-6R), but could inhibit the IL-6/IL-6 complex binding to gp130. 1656 INTLLSEINSILLDIISLL(2)[2.709 ± 0.168] INTLLMEINNILLNIIDFL(1)[3.088 ± 0.161] INSLLMEINTILNDIIMFL(1)[2.983 ± 0.185] INLLLMEINMILNEIITLL(1)[3.409 ± 0.061] 4 mRNA display (competitive panning) 8 PMID:21136209 Interleukin-6, IL-6 Interleukin-6 receptor 1P9M, Not determined. X(3)-L-I-X-E-I-X(2)-I-L-X(2)-I-X(3)-L mRNA display library ELISA In peptide ELISA, the absorbance at 490 nm was determined. Data were reproduced from the graph and expressed as the mean ± SD of triplicate determinations. The phage RNA(-) acted as negative control with A490 of 0.037. Beside, both RA07 (INTLLSEINSILLDIISLL) and CA11 (NQQLIEEIIQILHKIFEIL) peptides bound to GST-IL-6 in a dose-dependent manner and RA07 had higher affinity for GST-IL-6 (EC50=45.1 nM) than CA11 (EC50=142 nM). To determine whether CA11 and RA07 have inhibitory activity against IL-6, a KT-3 cell-based assay was performed. KT-3 cells are known to proliferate in response to IL-6. Results showed that RA07 suppressed KT-3 cell proliferation in a dose-dependent manner, but CA11 did not. The IC50 value of RA07 was 5.6 μM. Besides, the results of ELISA-based IL-6 inhibition assay suggested that RA07 inhibits gp130 from binding to the IL-6/IL-6R complex, but not IL-6R from binding to IL-6. The IC50 value of RA07 in this assay was 2.8 μM, which was almost the same as in the KT-3 cell-based assay (5.6 μM). After the beads were washed three times with selection buffer, 1–8 μM of IL-6 (Ajinomoto) was added and incubation continued another 2 h. The bound molecules were eluted with PreScission protease (GE healthcare). The most frequently occurring sequence, RA07 (INTLLSEINSILLDIISLL), bound to IL-6 with 3 to 4-fold higher affinity than CA11. Furthermore, RA07 inhibited IL-6-dependent KT-3 cell proliferation in a dose-dependent manner. ELISAs revealed that RA07 could not inhibit IL-6 from binding to the IL-6 receptor (IL-6R), but could inhibit the IL-6/IL-6 complex binding to gp130. 1657 INILIMEINNILKTIIREL(2)[1.252 ± 0.089] 1 mRNA display (competitive panning) 8 PMID:21136209 Interleukin-6, IL-6 Interleukin-6 receptor 1P9M, Not determined. X(3)-L-I-X-E-I-X(2)-I-L-X(2)-I-X(3)-L mRNA display library ELISA In peptide ELISA, the absorbance at 490 nm was determined. Data were reproduced from the graph and expressed as the mean ± SD of triplicate determinations. The phage RNA(-) acted as negative control with A490 of 0.037. The bound molecules were eluted with PreScission protease (GE healthcare). 1658 FPRWKLLAHWADRWWF(5) MHYEAVRVGRVLRFLA(4) IRMMVSFLRRQSRWWL(1) ARVFGLWLRSMSDMWL(1) MWVHSAAMATGLASRM(1) GNRWQYWIAFRMRYVA(1) RHLLLVDRLRAIALLL(1) RVLRYYLVGLALRQMA(1) AAVASMLRHLAGLFVL(1) FPRWKLLAHWADRGWC(1) FPRWKLLAHWADRWWV(1) FLRWKLLAHWADRWWF(1) SQRSTMYIAAVLRWLA(1) MVPSWVGLARVLRWLA(1) LVLLGRMLKFVA(1) 15 mRNA display (common panning) 5 PMID:20181936 B-cell lymphoma/leukemia 10 Not determined. Not determined. Not determined. X16 mRNA display library Fluorescence polarization To evaluate the affinity of the selected peptides for the BH3-recognition site of Bcl-XL, competition fluorescence polarization assay was performed. The selected peptide A10 (FPRWKLLAHWADRWWF) (IC50=0.9 μM) showed marked inhibitory activity. The selected peptides have sequence similarity with the Bcl-2 family BH3 domains, and one of them (FPRWKLLAHWADRWWF) has higher affinity (IC 50 = 0.9 μM) than Bak BH3 (IC 50 = 11.8 μM) for Bcl-X(L) in vitro. We also found that GFP fusions of the selected peptides specifically interact with Bcl-X(L), localize in mitochondria, and induce cell death. Further, a chimeric molecule, in which the BH3 domain of Bak protein was replaced with a selected peptide, retained the ability to bind specifically to Bcl-X(L). These results demonstrate that this selected peptide specifically antagonizes the function of Bcl-X(L) and overcomes the effects of Bcl-X(L) in intact cells. 1659 YQVARMLRRVADQMAS(7) KVVMRQLLMIADTMAR(1) LTVARRLKWVADIIFA(1) 3 mRNA display (common panning) 5 PMID:20181936 B-cell lymphoma/leukemia 10 Not determined. Not determined. Not determined. X(2)-V-X-R-X-L-X(4)-D-X-I-X(2) mRNA display library Fluorescence polarization To evaluate the affinity of the selected peptides for the BH3-recognition site of Bcl-XL, competition fluorescence polarization assay was performed. The selected peptide B01 (YQVARMLRRVADQMAS) (IC50= 6.4 μM) showed marked inhibitory activity. 1660 ALLSDLYKLL(1) DSASDLYKLL(1) ESDLFKMASS(1) FLYMSDLYKL(1) HTQFSDLFKL(1) LMHKARTTWP(1) LSDLSKLAQS(1) LSQNGLTSFN(1) MCNHSDLNKL(1) MWSDLTKLLP(2) NMSFSDMWKL(2) PEMSSDLWKL(1) PLYSDLLKLL(1) PQDSDLYKLS(1) RIWMSDLVKL(1) RLQDSAVSGH(2) SDLWKMMDKN(2) SHRMSDLWKL(1) SNYVSDLYKL(1) SVKAWPSAQI(1) SYTESDLHKL(1) TLRFSDLIKL(1) TSSDLWKLLP(2) TTHSIVSHVS(1) VYTSSDLNKL(1) WLVESDLYKL(2) YSDLDKMKPY(1) YSDLVKLSTL(1) x-S-D-L-x-K-L 28 mRNA display (subtractive panning) 4 PMID:18855146 Anti-p53 monoclonal antibody Cellular tumor antigen p53 Not determined. Not determined. X10 mRNA display library In order to remove non-specific binders to antibodies, the molecules prepared in the previous section were incubated with human normal IgG (Sigma)-bound Protein G-Sepharose beads (GE Healthcare) for 30 min at room temperature with rotation. Although no peptides that fully matched with hTP53 were found in the clones isolated, the core sequence was found in hTP53. A 10-amino acid peptide containing the core sequence was chemically synthesized to verify its binding with SPR. Its Kd value for the antibody was 6 nM. 1661 AEYSSDLWKL(1) DYSDLNKLMH(1) ERFSDLMKLA(1) FIDLYNLLMP(1) FPFMSDLYRL(1) FSDLDKLKAT(1) FSDLWKLSVVE(1) HGYSDLWKML(1) HNPFSDLHKM(1) KPKNSARLHQ(1) KTPYSDLWKL(1) LLESDLYKLL(1) LTFSYLWKLR(1) LWTFSDLNKL(1) LYSDLTKLQC(1) LYTSSDLWKL(1) MTTSDLWKLL(1) MYSDLDKLLI(1) NLHYNDLFKL(1) PELFSDLWKL(1) PFSDLDKLIP(1) PPSDLDKFNT(1) PYWSDLHKLA(1) QPLYSDLYKL(1) SDKLSDLHKL(1) STFSDLWKLA(1) SVYSDLHKLA(1) SYSDLYKLCT(1) SYYTDLDKLL(1) TSLSDLYKLS(1) YISDLWKLLF(1) YLWSDLWKLQ(1) YSDLNRLKNT(1) YSDLSKLLTL(1) x-S-D-L-x-K-L 34 mRNA display (subtractive panning) 5 PMID:18855146 Anti-p53 monoclonal antibody Cellular tumor antigen p53 Not determined. Not determined. X10 mRNA display library In order to remove non-specific binders to antibodies, the molecules prepared in the previous section were incubated with human normal IgG (Sigma)-bound Protein G-Sepharose beads (GE Healthcare) for 30 min at room temperature with rotation. Although no peptides that fully matched with hTP53 were found in the clones isolated, the core sequence was found in hTP53. A 10-amino acid peptide containing the core sequence was chemically synthesized to verify its binding with SPR. Its Kd value for the antibody was 6 nM. 1662 DATVCSRVWSSHC(1) ECARVVPGLMGYC(1) ECAVDVFVYMVSC(1) ECCEAYHLYLTPC(1) ECCGISRGVRTXC(1) ECCLWKSGHGSMC(1) ECDRRCKFKFRAC(1) ECGARGLECAGAC(1) ECGFLTEADCGSC(1) ECGHGSRGGPGCC(1) ECGSDGRELRYGC(1) ECKLHGGSCSCAC(1) ECLKKIEVSQLSC(1) ECQCRIFSKRSSC(1) ECRKPRGGYRDAC(1) ECRLLRGHGEARC(1) ECRRVPLAPQGSC(1) ECTLWDSREQPTC(1) ECVEGIDANPVAC(1) ECVLVAWFDVREC(1) ECWCIIRPVEPCC(1) ECWEDLVDDWRDC(1) GAGERFNVWC(1) GCAFARXLXKVFC(1) GCATKMVGAPRCC(1) GCCGGVRHTGFTC(1) GCCGSGFSLEEVC(1) GCDRGVELVNGVC(1) GCESVSRTPHAGC(1) GCFHVRMLTARGC(1) GCGAAREGGADRC(1) GCGGILKVWLVAC(1) GCGHGAAADALHC(1) GCIVALWLVFQIC(1) GCLGFDDQGIGRC(1) GCLSRVCVRYSAC(1) GCLVGDLGACAVC(2) GCMFCRSGDFASC(1) GCNVPTCQAPRRC(1) GCNWLILDAAFSC(1) GCQQEHGCHRYSC(1) GCRGGADYTTSYC(1) GCRNSGVSAGGKC(1) GCRSRSMLSMRMC(1) GCSAWADLELDTC(1) GCSRGKKKWVIRC(1) GCSTTDIQGLLYC(1) GCSVAGLLHAVGC(1) GCTALEGGRTAHC(1) GCTPRSLDSYGGC(1) GCTVGCRNAAEPC(1) GCVDPCTCVRPGC(1) GCVGGGTLSSLNC(1) GCVIEGRYFRCRC(1) GCVLAITRRSVWC(1) GCVLFVSTREYVC(1) GCVRAIVFGIYIC(1) GCWMAVGNHEVSC(1) GCWTDVSYWSFFC(1) GCYRWPVEKPLDC(1) KCALVFRSDGVLC(1) KCCRTLQHAVWRC(1) KCDGVECVAVEKC(1) KCFLTSGSWVSAC(1) KCGWNAIKSEGGC(1) KCHGCTCREWDHC(1) KCNFAVLSGDLVC(1) KCSRGIRCAGVLC(2) KCVRNLANGGAVC(1) KCWLCYGGLYNYC(1) KCWLSFAPYAGCC(1) RCATIRAVYAHSC(1) RCCLDVVFWQVPC(1) RCDASHALKKLHC(1) RCDGNRVVFAVIC(1) RCFSQASRDAEVC(1) RCGACGHVCRYTC(1) RCGAWQMLLLLGC(1) RCGCGRSSSLFNC(1) RCGSREGLSFLQC(1) RCIVNGPVTRILC(1) RCLQSRWDGFPWC(1) RCLRAGLPCSNSC(1) RCLSALTCKMALC(2) RCLTGTFAPWAFC(1) RCMLILRCYSAGC(1) RCRASGAGACMLC(1) RCRHRFLRFVASC(1) RCRLEPPDCRTRC(1) RCRLYDMLAVCLC(1) RCRQIGCALVGQC(1) RCRSSQTATASRC(1) RCRWGGVSGFFLC(1) RCSQIAVSIIGPC(1) RCSTRAEAAREGC(2) RCTCLGLHHFIKC(1) RCVGRVDKNKFLC(1) RCVGVIWDSEARC(1) RCVHSSTAVWFRC(1) RCWGTRMGQRGTC(1) 100 mRNA display (competitive panning) 6-7 PMID:18824687 Internal ribosomal entry site, IRES Not determined. Not determined. Not determined. [RKEDG]-C-X(10)-C mRNA display library Rounds 1 and 2 were designed to decrease the complexity of the starting library while retaining all possible IRES binders. During these rounds Torula yeast RNA (TYR) was used as a binding competitor, and all column matrix bound material was eluted nonspecifically with NaOH and 8 M urea. Rounds 3-7 were designed to select more specifically for IRES binders by eluting column bound mRNA-peptide fusions competitively with soluble IRES. Following seven rounds of selection, ≈50% of the input library was eluted from IRES column with soluble IRES RNA in 2 h. 1663 GCGSCPVCHGYPC(1) GCGGTEGHIGRGC(1) GCCWTAAMAGTSC(1) KCSRGIRCAGVLC(15) KCSRGIRCAGVRC(1) KCSRGIRCAGVLX(1) GCTACDRMYLVCC(1) GCRVCNRMEGVLC(1) GCLYLWRQGGLAC(1) RCIDHQYSWLCYC(1) ECGCSHPVFLCYC(1) RCWESYVDHLDLC(1) GCQSYGPGDLCLC(1) GCVGYSRIGLSVC(1) GCWISISACFKSC(1) GCVYSRLKVFADC(1) GCWVEMRGSWKRC(1) GCWGCMLSPEFRC(1) ECIASSGRAGGVC(1) GCRAASSVSWMWC(1) KCKKPGSARWSKC(1) RCNKSLIAIRYDC(1) ECGYRKIARMVMC(1) RCPSEGYHRRTGC(1) ECCQSGRPRDGGC(1) GCSGVGAARTWGC(1) GCTFGTQPRHWCC(1) ECHVTQHPYSRAC(1) KCKGFVGFFSRAC(1) ECTDCYLALSSYC(1) ECRRCFAELSYAC(1) ECLCYSYAGSCRC(1) KCTLFKAAGGPFC(1) RYGSFDDMSCGEC(1) GCYDRMPGGTHSC(1) GCGLGRARTDVAC(1) RCEGCVHYMGLSC(1) GCDRWSAGCVVC(1) RCAFCDFLTRXLC(1) 39 mRNA display (competitive panning) 11 PMID:18824687 Internal ribosomal entry site, IRES Not determined. Not determined. Not determined. [RKEDG]-C-X(10)-C mRNA display library Rounds 1 and 2 were designed to decrease the complexity of the starting library while retaining all possible IRES binders. During these rounds Torula yeast RNA (TYR) was used as a binding competitor, and all column matrix bound material was eluted nonspecifically with NaOH and 8 M urea. Rounds 3-7 were designed to select more specifically for IRES binders by eluting column bound mRNA-peptide fusions competitively with soluble IRES. Following seven rounds of selection, ≈50% of the input library was eluted from IRES column with soluble IRES RNA in 2 h. Authors then further increased the stringency of the selection by using additional selection steps in rounds 8-11, aimed at the selection of peptides with slower dissociation rates, and improved IRES selectivity. Cloning and sequencing of the library after round 11 revealed that the sequence 6B4, encoding the peptide MKCSRGIRCAGVLCGSVGHHHHHHHRL, accounted for >30% of the clones. The best peptide binds the IRES with subnanomolar affinity, and a specificity of at least 100-fold relative to binding to several other RNAs of similar length. The peptide specifically inhibits HCV IRES-initiated translation in vitro with no detectable effect on normal cap-dependent translation initiation. 1664 HCYPLDESILRLVRGELGAVH YWFAWAKSWQLAKREGHMGFD NWSGFYNLKLNTLKSVRQVHY NWHHALLSDIVSALNKSSLWN NCGTEMFCMDKINEWREVYVD NWSLIHKKYLQQAKMKYARCH DWTLMKVHVKQCVRKLESFTN HCRFVRRVAGLMFRERLEDDH NWRLSHNAVLDDARVQYGVHH 9 mRNA display (common panning) 12 PMID:17879061 Thymidylate synthase mRNA, TS Not determined. Not determined. Not determined. YAC-TGZ-(XYZ)18-YAC mRNA display library Comparing the amino acid composition of the selected peptides (12th round, R12) with those from the initial random library (round zero, R0), the basic and aromatic residues in the selected peptides were enriched significantly, suggesting that these peptide regions might be important in the peptide-TS mRNA interaction. Categorizing the amino acids at each random position based on their physicochemical properties and comparing the distributions with those of the initial random pool, an obvious basic charge characteristic was found at positions 1, 12, 17 and 18, suggesting that basic side chains participate in RNA binding. Secondary structure prediction showed that the selected peptides of R12 pool represented a helical propensity compared with R0 pool, and the regions were rich in basic residues. The electrophoretic gel mobility shift and in vitro translation assays showed that the peptides selected using mRNA display could bind TS RNA specifically and inhibit the translation of TS mRNA. 1665 NWENKSDVLNKMLHLRLITN(1) HWEKIRDNVIHKSDRSYYQKH(1) NWKSIIQRNLRWNKFKRFYQD(1) NWNILRQEVMKMGPAKDTVRN(1) NWNTVWSAYRRASSKLTKKTY(1) NWNRWRWIYLAQKNHKECLKN(1) YWKRYRLMALKANFGSRYSRN(1) NCPSTRKRWIVYLYRGRLINN(1) NWNRVRRYCKCKYTLVAKNNY(1) NWYTVKQMMIKDRRRSQAMYN(1) NWSDIRMHGLMRRREKNVLNH(1) NWVKLRQRVTLAKRVAVNLNY(1) HWNKVASERINKMRRKAILNY(1) HWNAIKNVVHSQRMTKKIAMN(1) NWEILRSKYNSHMTKSNVFTY(1) YWSELRRKKWWKVWKLSSCPH(1) YWNHELRVRLINRIKMTN(1) HWGIIRQKIVVAHDIFQCKDY(1) NWINNVRLRIHTKRWLLKSNH(1) NWLRLVPRIKALNKVQVKNHN(1) NCKMQPQNWYHVYRMSRLVKN(1) DCEYRDKVTLFNLVRLVMTKN(1) YWSNRVTQSIKARYVIDSWQD(1) NWHKVFIRRQSKKLVYNTIKN(1) YCHKYTVANESHWSKIRLKMY(1) YWVFLKKKMKLENKCVRVVKN(1) DWNMLKVKLYALRVRRRRMAN(1) HWMRTSQRVRVNNAFHKYMGY(1) NCEVLKTQRWRKVLQQHIIRH(1) HWSKTQGGNKRWRMIGAVVAH(1) NWDKVRSTFKKCHSIVIFKRN(1) YWSLIVSKLRRRKVMNDPSTY(1) HWDRVRLTMLRSRLKDDKKKH(1) NCLKQRLLRTPYLMMSRAVTH(2) NWRKAINLVRKWRNNDDPNKD(1) HWAMTRWHILANNVMNRRTCD(1) YWQRLQSMLKRVDPRPYRVRD(1) HWNKLSYTVRIGEIKRYVWRN(1) YWRQKAKDDLMVRRLRRAVKH(1) NWRNLRLKWRRLNSVHRH(1) 40 Phage display (common panning) 2 PMID:17685586 Calmodulin, CaM Not determined. Not determined. Not determined. X20 mRNA display library The mRNA display library passed through a precolumn with streptavidin beads to minimize the enrichment of matrix-binding sequences. The flow-through was incubated with biotinylated CaM. Molecules that bound to CaM in a Ca(2+)-dependent manner were eluted using the same buffer containing 2 mM EGTA. More than 2000 CaM-binding peptides were selected from the combinatorial peptide library. Unlike sequences from prior CaM-binding selections that correlated poorly with naturally occurring proteins, synthetic peptides homologous to the Ca(2+)/CaM-binding motifs in natural proteins were isolated. Interestingly, a large number of synthetic peptides that lack the conventional CaM-binding secondary structures bound to CaM tightly and specifically, suggesting the presence of other interaction modes between CaM and its downstream binding targets. 1666 MMFLRCRATKML MMLPCCRATKML MVCCACRATKML MSYCWCRATKML MSLVVCRATKML 5 mRNA display (common panning) 1 PMID:16122707 Botulinum neurotoxin A light chain Not determined. Not determined. Not determined. M-X(4)-CRATKML mRNA display library Purified biotin-ubiquitin BoNT/A LC immobilized on Softlink monomeric avidin (Promega) was first incubated with the RNA-peptides for specified time. RNA-peptide fusions bound to the immobilized BoNT/A LC were eluted off the column with 5 mM biotin in the same buffer. 1667 MVMLACRATKML MWMTSCRATKML MAFFGCRATKML MLLGGWCRATKML MVLVGCRATKML MFRLACRATKML MLTPRCRATKML MLVPACRATKML MGSAYCRATKML MGLLCCRATKML MGLRFCRATKML 8 mRNA display (common panning) 2 PMID:16122707 Botulinum neurotoxin A light chain Not determined. Not determined. Not determined. M-X(4)-CRATKML mRNA display library Purified biotin-ubiquitin BoNT/A LC immobilized on Softlink monomeric avidin (Promega) was first incubated with the RNA-peptides for specified time. RNA-peptide fusions bound to the immobilized BoNT/A LC were eluted off the column with 5 mM biotin in the same buffer. A protease assay was developed using immobilized intact SNAP-25 as the substrate. The new peptide inhibitors showed a 10-fold increase in affinity to BoNT/A light chain than the parent peptide. Interestingly, the sequences of the new peptide inhibitors showed abundant hydrophobic residues but few hydrophilic residues. 1668 MVMLACRATKML(3) MWMTSCRATKML(1) MAFFGCRATKML(1) MPGARCRATKML(1) MLGQHCRATKML(1) MLRGFCRATKML(1) 6 mRNA display (common panning) 3 PMID:16122707 Botulinum neurotoxin A light chain Not determined. Not determined. Not determined. M-X(4)-CRATKML mRNA display library Purified biotin-ubiquitin BoNT/A LC immobilized on Softlink monomeric avidin (Promega) was first incubated with the RNA-peptides for specified time. RNA-peptide fusions bound to the immobilized BoNT/A LC were eluted off the column with 5 mM biotin in the same buffer. A protease assay was developed using immobilized intact SNAP-25 as the substrate. The new peptide inhibitors showed a 10-fold increase in affinity to BoNT/A light chain than the parent peptide. Interestingly, the sequences of the new peptide inhibitors showed abundant hydrophobic residues but few hydrophilic residues. 1669 YRTNHHYDVGRFAARGRRD(1) NGRSSMNWRSQEITRYTSEHHYRMAFL(1) PEQYDHHHLEARRRASSTRQVRARARR(1) RAYTPHHHAEGRLVRLEPHPAPYKNRT(1) YYVKNRLHHHRLARLVAAEHAHRLRVQ(1) NKRNLSYPWSHHHQVARRTHMRAQHTM(1) RPTKNFEAEVVRSTGPMHHHDTAKQRY(1) DFLTYNKSMGGRPTNFRHHHSSVVQSQ(1) DEPEVVGRVLGERPAGALADHHHMMKW(1) EVLHGHHHVVARVRASCTGPTRRASCA(6) HVYEKANNRLGHKHHHLAARRRSKSWN(1) SNKGFSWRKKGMAVTPNRHLHHHMVAH(1) TNHRHHHGVLERRQDILTGSLIEHKH(1) ILKRLREQHRHHHAAAHHVRVRRRGRH(1) NYTTRRAEWNRQDAHRHHHQEARRGAL(3) SKKDNAVGLQELRLREGHRHHHDVMLT(1) KKVRGHHRHHHQVALLDAAERGPGRMS(1) GIHHHHAMAVLAELGMNPMGFALPDMW(1) AGVHHHHDAARGGTRSRRSTPRSATRR(1) TMNWHHHHENGLRARMYDAGRR(1) KVRRDVMRWHHHHRMARRKANR(4) RVQDRLGHRAVQPVLHHHHQAARRRVR(1) AALHHHHHDAGRASAMRRPGTPATSWR(1) DGHPERHDAGDHHHHHGVRQWRLISTG(20) 24 mRNA display (subtractive panning) 6 PMID:15980016 Anti-polyhistidine monoclonal antibody HIS-1 Recombinant polyHistidine tagged fusion protein Not determined. Not determined. X27 mRNA display library The reverse-transcribed fusions were pre-cleared by rotating with 20 ml of protein G-Sepharose for >1 h. Bound RNA-peptide fusions were eluted with acetic acid through a 0.45 mm spin filter (SpinX, Costar). After six rounds of selection, epitope-like peptides were identified that contain two to five consecutive, internal histidines and are biased for arginine residues, without any other identifiable consensus. The epitope was further refined by constructing a high-complexity, unidirectional fragment library from the final selection pool. Selection by mRNA display minimized the dominant peptide from the original selection to a 15-residue functional sequence (peptide Cmin: RHDAGDHHHHHGVRQ; K(D) = 38 nM). 1670 MSQTKRLDDQLYWWEYL 1 mRNA display (subtractive panning) 6 PMID:15248784 Guanine nucleotide-binding protein G(i) subunit alpha-1 Not determined. Not determined. Not determined. GPR X(6) mRNA display library Surface plasmon resonance (SPR) The dominant peptide from the selection, R6A (MSQTKRLDDQLYWWEYL), retain high affinity (KD = 60 nM) for the GDP-bound state of GiR1, as measured by surface plasmon resonance. The affinity matrix for selection was prepared by rotating Nb- and/or Cb-GiR1(~10μg each) with ~20μL streptavidin agarose (Immobilized NeutrAvidin on Agarose, Pierce) in buffer A (20 mM HEPES-KOH at pH 7.5, 200 mM NaCl, 5 mM MgCl2, 1 mM EDTA, and 0.05% Tween 20) at 4 °C for >1 h. The slurry was supplemented with 1 mMD-biotin (~0.1 mM final concentration) and rotated for an additional 10 min to block biotin-binding sites. After washing thoroughly with buffer A2 [buffer A supplemented with 2μM GDP, 1 mM β-mercaptoethanol, 0.2% (w/v) BSA, and 1μg/mL yeast tRNA (Roche Diagnostics Corp., Indianapolis, IN)], reverse-transcribed fusions were rotated with the affinity matrix in 1 mL of buffer A2 at 4°C for 1 h. A total of 6 rounds of selection were performed on a mixture of immobilized Nb- and Cb-G iR1 to reduce the effects of bias or steric hindrance with either terminus immobilized. Detergent, bovine serum albumin (BSA), and salt were included in selection buffers to minimize recovery of nonspecific binding peptides. DNA sequencing of the 6th round pool revealed a dominant peptide sequence (MSQTKRLDDQLYWWEYL). Selected peptides contain mutations to a highly conserved Arg in the GPR motif, previously shown to be crucial for binding and inhibition activities. The dominant peptide from the selection, R6A (MSQTKRLDDQLYWWEYL), and a minimal 9-mer peptide, R6A-1, do not contain Arg residues yet retain high affinity (K(D) = 60 and 200 nM, respectively) and specificity for the GDP-bound state of G(iR1), as measured by surface plasmon resonance. 1671 MSQTKRLDDQLYWWEYL MSQSERLDDQWTWWEFL MSQSKWLDDQLTWLEFL MSQSKRLDDQLTWLEFL MSQSKQLTITEFLQWL MSQSKRLEITWWEFVE 6 mRNA display (subtractive panning) 8 PMID:15248784 Guanine nucleotide-binding protein G(i) subunit alpha-1 Not determined. Not determined. Not determined. GPR X(6) mRNA display library Surface plasmon resonance (SPR) The dominant peptide from the selection, R6A (MSQTKRLDDQLYWWEYL), retain high affinity (KD = 60 nM) for the GDP-bound state of GiR1, as measured by surface plasmon resonance. The affinity matrix for selection was prepared by rotating Nb- and/or Cb-GiR1(~10μg each) with ~20μL streptavidin agarose (Immobilized NeutrAvidin on Agarose, Pierce) in buffer A (20 mM HEPES-KOH at pH 7.5, 200 mM NaCl, 5 mM MgCl2, 1 mM EDTA, and 0.05% Tween 20) at 4 °C for >1 h. The slurry was supplemented with 1 mMD-biotin (~0.1 mM final concentration) and rotated for an additional 10 min to block biotin-binding sites. After washing thoroughly with buffer A2 [buffer A supplemented with 2μM GDP, 1 mM β-mercaptoethanol, 0.2% (w/v) BSA, and 1μg/mL yeast tRNA (Roche Diagnostics Corp., Indianapolis, IN)], reverse-transcribed fusions were rotated with the affinity matrix in 1 mL of buffer A2 at 4°C for 1 h. The first 6 rounds of selection were performed on a mixture of immobilized Nb- and Cb-G iR1 to reduce the effects of bias or steric hindrance with either terminus immobilized. Detergent, bovine serum albumin (BSA), and salt were included in selection buffers to minimize recovery of nonspecific binding peptides. An additional 2 rounds of selection was preceded by a subtractive hybridization step. Selected peptides contain mutations to a highly conserved Arg in the GPR motif, previously shown to be crucial for binding and inhibition activities. The dominant peptide from the selection, R6A (MSQTKRLDDQLYWWEYL), and a minimal 9-mer peptide, R6A-1, do not contain Arg residues yet retain high affinity (K(D) = 60 and 200 nM, respectively) and specificity for the GDP-bound state of G(iR1), as measured by surface plasmon resonance. 1672 MLRNSNCIRHFFGVDYKDDDDKGGGG(2) MDRTPTCTFLSSGGDYKDDDDKGGGG(1) MNFANVCVSQHIGGDYKDDDDKGGGG(1) MATKTDCFLSLVGGDYKDDDDKGGGG(1) MNARWNCNSWLVGGDYKDDDDKGGGG(1) MGRHTPCVSNLYGGDYKDDDDKGGGG(1) MSPYRSCGLSASGVDYKDDDDKGGGG(1) MRNNITCRLLKRGVDYKDDDDKGGGG(1) MRACTTCSWPLSGVDYKDDDDKGGGG(1) MRHNLNCSAFWPGVDYKDDDDKGGGG(1) MSLSSICLVPVAGATTRTTTTRAAA(1) MESHMVCRSTDVDYKDDDDKGGGG(1) MSPAHCCPYLPFDYKDDDDKGGGG(1) MSLAAHCAFPFLDYKDDDDKGGGG(1) 14 mRNA display (common panning) 9 PMID:12670537 Penicillin binding protein 2a, PBP2a Not determined. Not determined. Not determined. X(5)-C-X(5) mRNA display library A fixed cysteine residue in the mRNA fusion library (X(5)-Cys-X(5)) reacts with sodium 6-bromoacetyl penicillanate to form a peptide-drug conjugate. In vitro selection using this hybrid peptide-drug library resulted in novel inhibitors of the Staphylococcus aureus penicillin binding protein 2a (PBP2a). This strategy resulted in a penicillin-peptide conjugate (LRNSNC[Pen]IRHFF) that has at least 100-fold higher activity than the parent penicillin itself. 1673 ERNYNDFCDPGRVGL(8) YFCMGQICFLRFALH(2) DFDNMFSNLDPGRHW(2) FFDRYDSARDPGRLL(2) ERNYNDFCDPGRGGL(2) ERHNDPGRYFVEYET(2) FYESCENSDPGRTYA(1) FHESCENSDPGRTYA(1) YTHSSKSSDPGRKLW(1) DYSNESSDPGRLWHC(1) ERNYNDFCDPGRFGL(1) ERNYNDFCNPGRVGL(1) ERNYNDLCDPGRVGL(1) WRHYNPSDDPGRVHL(1) EYTNDYDDPGRTLSG(1) RNYTNPYCDPGRQHE(1) HGPDVNHADPGRYFD(1) LHLDSDHFDPGRTVW(1) LVDCISHDDPGRSVG(1) DINYCDPGRDCDGHL(1) DSYCELTDPGRWISA(1) EPCCCENRDVGRLIH(1) EFNQWEDPGRMRVGC(1) NHNLMNDPGRFFWHD(1) YTYSNDFTDGGRHIL(1) DYVSDVCRDGGRIML(1) YFDPGFCVFTSDHLA(1) YDAGFCNYDRDHIWP(1) YNSLSGRSLHPDIGF(1) DNYKLCESDVGRLLF(1) WFVYIDRWAYASFRH(1) YVYLLRHDQHSYYPP(1) YLHVVEIERHRIRFF(1) WEIYWHLEFAGFDRV(1) YALIYAVKKRMGIAH(1) WQRCGMAEFIWHGQW(1) YGHCFENFGDSFEHN(1) DYWAFWRVYFQVDGY(1) LYVRRSTAHIFVYAN(1) WLVCRHSKRYNCIFL(1) LRLVGSDHNFDAVVC(1) YMSFTRRTESDKLHS(1) DRSPRMFYNRFNSAL(1) LNWPNNLEGRDPYNR(1) DRSILVPRYFELWAN(1) DPGR 45 mRNA display (common panning) 10 PMID:12573700 Prothrombin Not determined. Not determined. Not determined. X15 mRNA display library Surface plasmon resonance (SPR) Thrombin was used as analyte at increasing concentrations (1-2500 nM). Dissociation constants were then calculated by the BIAevaluation software using steady-state affinity curve fits. Kd values of 520 ± 6 nM for T10-11 (ERNYNDFCDPGRVGL) and 166 ± 18 nM for T10-39 (FFDRYDSARDPGRLL) were determined. Bound peptides were eluted with elution buffer (4 M urea, 0.5% SDS). Two clones (FFDRYDSARDPGRLL and ERNYNDFCDPGRVGL) showed binding affinities ranging from 166 to 520 nM. A conserved motif of four amino acids, DPGR, was identified. Clot formation of human plasma is inhibited by the selected clones, and they downregulate the thrombin-meditated activation of protein C. The identified peptide motifs do not share primary sequence similarities to any of the known natural thrombin binding motifs. 1674 MDAQTRRRERRAMERAQTSSSL(1) MDAQTRRRERRAMERATLPQVL(11) MDAQTRRRERRAALRNEKFWVV(1) MDAQTRRRERRALMRRNARIAV(1) MDAQTRRRERRARDRFRLLRTL(1) MNAQTRRRERRALAFKARYMAL(1) MDAQTRRRERRAWERRTQSWMV(1) 7 mRNA display (common panning) 11 PMID:11675487 Three RNA targets (λ-boxBR, GNRA tetraloop, and P22 boxBL) Not determined. Not determined. Not determined. X10 mRNA display library Each peptide tested could bind at least one of the hairpins in the mixture. The specificity of seven individual clones, the round 11 and round 12 pools were then tested by using each individual hairpin. All showed a marked preference for the boxBR RNA as compared with the GNRA or P22 targets. 1675 MDAQTRRRERRAWKTVLWLQAL(1) MNAQTRRRERRAMEREERSKTV(1) MDAQTRRRERRALERTKLEKAL(1) MDAQTRRRERRALQRSRARHAL(1) MDAQTRRRERRAMERLMHERQA(1) MDAQTRRRERRAHERKISFTAL(1) MDAQTRRRERRAELRKSSLAFL(1) MDAQTRRRERRAWERTQRKNEY(2) MDAQTRRRERRALSRKMAIRIL(1) MDAQTRRRERRANIRRQSAWVL(1) MDAQTRRRERRALERQKSLRAL(1) MNAQTRRRERRAKDLKNTGMPF(1) MDSQTRRRERRAVRRESQYGSL(1) MDAQTRRRERRANMRMYRSLVI(1) MDAQTRRRERRAQSRMTNSWVL(1) MDAQTRRRERRATMRDSRSLAL(1) MDAQTRRRERRALERSMKLHAL(5) MDAQTRRRERRALDREMRGMVL(3) MNAQTRRRERRATLREERRVSW(3) 19 mRNA display (common panning) 12 PMID:11675487 Three RNA targets (λ-boxBR, GNRA tetraloop, and P22 boxBL) Not determined. Not determined. Not determined. X10 mRNA display library Each peptide tested could bind at least one of the hairpins in the mixture. The specificity of seven individual clones, the round 11 and round 12 pools were then tested by using each individual hairpin. All showed a marked preference for the boxBR RNA as compared with the GNRA or P22 targets. 1676 PRILMR(0.2) PRLLAP(0.075) PRILLP(0.0375) PRLLQP(0.0375) PRVLAL(0.0375) PRVLAP(0.025) PRLLVL(0.025) PRLLHL(0.025) PRLVQR(0.025) PKLLAP(0.0125) P-R-x-L-x(2) 10 Phage display (subtractive panning) 5 PMID:12203838 Cationic trypsin Trypsin inhibitor 2 Not determined. Not determined. EETI-II mRNA display library (X6) Potential column binders were removed from the reverse transcribed EETI-II fusion library (1 pmol in 700μl) by incubating the library with 100μl CL-sepharose beads (Sigma) for 15 min at room temperature. The cDNA pool from the fifth round of selection was cloned and 108 clones were sequenced. The sequence analysis revealed that wild-type EETI-II represented 20% of the clones. The binding efficiency of each clone was determined and, unexpectedly, a lower percentage of wild-type bound than any of the other screened clones; despite the fact that the wild-type clone bound the tightest, followed closely by the P(1)-lysine variant. 1677 CASVISEREC(1) EEYLVSEYVM(1) RQYLISEYEH(1) LQRLISEQMF(1) IVRLLSEYHM(1) EEYLLSEYVM(1) MQNLISEHEL(1) TMDLIPEHYM(1) EQKLISEEDL(1) DMMLISEKEL(1) FQALIAEEEL(1) QRVLISEFWL(1) x-[QE]-x-L-I-S-E-x(2)-[LM] 12 mRNA display (common panning) 5-6 PMID:12203838 Anti-c-myc monoclonal antibody 9E10 Myc proto-oncogene protein Not determined. Not determined. c-myc mRNA display library (X27) The cDNA pools from the fifth and sixth rounds were cloned and 116 colonies were sequenced. As presented in Table 3, two selected clones contained the sequence of the wild-type c-Myc epitope (EQKLISEEDL). A third clone differed from the wild-type by two point mutations in the nucleotide sequence, only one of which altered an amino acid (Ile to Val). The consensus sequence selected [X(Q,E)XLISEXX(L,M)] included the full length 10 amino acid wild-type sequence; identical or homologous sequences to the four of the core residues, LISE, were conserved in 86% of the 116 clones examined. Experiments confirmed the specific binding of the RNA-peptide fusions to the antigen-binding site of the anti-c-Myc monoclonal antibody. The selected sequences bound the c-Myc antibody with an affinity similar to that of the wild-type Myc fusion. 1678 LYDIDRNWVGHPQG(5) NYDADLAWDTHPQD(4) MHSDLDNMWETHPQ(1) ITDWLSHPQGPRSN(1) MYSADDTFRHMHPQ(1) CFNGQEDRINHPQG(1) DVGSWSGNEFIHPQ(1) DLSSPWLVGHPQAT(1) YWNSNDMAMHPQVN(1) SINLMDRFASHPQA(1) VEAWLDERVPLVET(3) VEAWLADRSQVSPN(1) VCAWLEVESHPVRE(1) VLSWLESNSVRITR(1) VEAWIADPAVHFTT(2) VEAWISYQSPLPDQ(1) VEAWIMHPCPLCSC(1) VESWVEQYSVPIVP(1) VNAWVSNMDCKECR(1) LYKVPSHCHPMMPC(2) EYLSGVQFATWQCP(1) TWTVGCSCMMACVH(1) CLAVLQNEVHPVYD(1) GMTEWWYGLRCVVA(1) SFNGFQNEGHPLDD(1) HPQ, DVEAW, D-V-x(2)-W 25 Ribosome display (common panning) 7 PMID:12758084 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. [VGAED]-X(14) ribosome display library The 37 randomly chosen clones were sequenced: 26 different sequences were observed and 46% of the sequenced clones contained the HPQ motif. Of the clones sequenced, 32% carried the unknown DVEAW or DVXXW motif. The remaining 22% of the sequenced clones revealed no further motif. 1679 RQERSSLSKPVV(12) 1 Ribosome display (common panning) PMID:16269342 Prepilin Not determined. Not determined. Not determined. X12 ribosome display library The random ribosome display system was used, and mRNA-attached peptide products bound to prePilS-GST-Sepharose 4B beads. An unmeasureably small fraction of total mRNA product was bead-associated. A 12-mer peptide (RQERSSLSKPVV), binding to the structural protein PilS of the type IVB pili of S. Typhi, was isolated with a ribosome display system. This peptide was designated as peptide R. Authors found that peptide R inhibited adhesion to/invasion of human monocytic THP-1 cells by piliated S. Typhi bacteria, but had no effects on nonpiliated S. Typhi bacteria. 1680 VWAWVFGASTRERARVGWQGY(1) CFVLPGVRPCSSHILTLSFSY(1) EGVRDMFRRCLWISLRSWCVH(1) GGVYASCASYLLALRSRVGGN(1) SDSASVSRVGGLWPTCCPH(1) WGRVDNSGSWGRVGAPWRYLH(1) CAAVYLVTSLFGIVTGVREDH(1) DSRGVRAFACDYVLFVLWVPY(1) DLAPVRVGFYNALRELRVFRY(1) RVGALRYFMVWYLMWFFLLFH(1) PVLDAGSVYLGYLGVRFLSY(1) VRNRVIARVGGVPYVGGPCYN(1) ACCLVRFYSHGRGKRVGFLWY(1) LRYSGLLGFPLWVGRIFVCVD(1) RMRCVSLELVVYGGGVRMWEN(1) FMGYGRSVWVVSSSLVLCIYD(1) ARMLWGRGTTLLLIRRRVSAY(1) VDLSWYASCRVSICVFVVVY(1) WPNYQSREHMALRSRMYYYFY(1) CVVALRNVKAAALIPGVVSRH(1) WWCLLGYWALGGNHSAALRSY(1) YGSYLEALRWGTSACWALRY(1) GVAVDCAVVGWALRVLGVHSY(1) RCLEAGKIWWGALRSHLAVYD(1) GSGSAVGWALRSYASGLAIAY(1) HAWARWMGWGHGGVLSWALRY(1) FVSWALRYSRCLVWLCWFPNY(1) VKGNPVFDHRHFSLWGALREY(1) FVQHWSFTAGSRSDRAPYPGH(1) AGWVNALRMWSLMPLMWLWSY(1) DRTTGRWFYIRRTAEVLGWTY(1) RFINPTSHCFGSLSLWRQLSY(1) VGCLVSVGSVWGCSSVVVRVY(1) RMESGAPLAAYGKMRLRPGTH(1) VWNRVIARVGGVLYVGGPCTN(1) SGHMHSYWPTTWILVLIRRTY(1) LEVLVWYSLWSYWLDVAAASH(1) SWGGGFYDWSYVGGGAYWAY(1) MGLFRSYKYRFVHDSESSFN(1) MALYLAWYGCSDSAVVMLADD(1) MYCWRMLANSCALRMVLAMRN(1) VIVNVAVLYRRCWPCAEFWPY(1) RLGSFYPLLWRLVSHEYSLWH(1) RYWFGRWRCFYGPFVSSYFLY(1) VCCCRCLPWSYMCEWGSMRLY(1) VLKIHSWHNWVYGVMLYDMEY(1) MGYAWDLALRMGPYFLMDLIN(1) SDKCAPVCYVMDRLCLANWD(1) ALR, RVG 48 Ribosome display (common panning) 5 PMID:16878349 Model membranes Not determined. Not determined. Not determined. X20-[YHND] ribosome display library Following stabilization of the peptide-ribosome-mRNA complex by addition of 50 mM Mg(2+), it was incubated with the model membrane immobilized on magnetic beads via streptavidin. mRNA was eluted from the bound complex in buffer containing EDTA. Of the resultant colonies, 50 clones were sequenced, 48 of which encoded 21 amino acids without a deletion. Authors obtained a high frequency of motifs consisting of several specific amino acids (e.g. ALR, RVG) in the peptides obtained from the random peptide library. These motifs consisted of a basic amino acid, K or R, and several hydrophobic amino acids, and may represent the minimum units essential for interaction with the bacterial membrane. 1681 RPVFRTYRSVVKSG(1) NRACLKRPRYLRKH(1) KACVRFSKSTSKRY(1) RFRPKAVRYRIKFN(1) RSHFRRVKRHSKTP(1) KDVLRNHKHSDRVG(1) KSARKVLKLYRKIT(1) KKNSCRDGRFSRKC(1) KGYFRGRRSYLRAF(1) KGCAKVLKRITRHI(1) KGRHRHCRYILRGN(1) KCTFRRRVLIIKPS(1) KTDWFKVLMTFLMD(1) RGFVRLIKPYAEAS(1) RNLCRSLRSHLEA(1) KIPGRFTRAGRKTT(1) KSDHKVLKNLPKTI(1) RRTGRIDKVSVKAY(1) KVLIKLAKCCIRIS(1) RAACRDSKLCSRYY(1) KHFVRCPKCAVRSS(1) KISDRNSKHHCRSS(1) KVGLIVDKASVKTA(1) RDVCKSSRHSHKGS(1) RFVSKGTDAINRRS(1) KGNCRLYRLRCKVV(1) RLLLKAVRFCCKCF(1) KGGGKVGKHTRSR(2) RHFRKNCKFCHRHC(1) KRCTKVLRAYTKLT(1) KSYGKAPKFVGRIC(1) RAAIRHFRSATKRP(1) KYSARFCKYGGRSH(1) RFTARVRKSVFRSC(1) KVYSRSSKSAHKCF(1) KRAYKDARHIYLCS(1) KIFVRTIRAAHKRD(1) KSLTKCCKVLRLSC(1) RCDIKSVKHILRCS(1) KASVRNSKVLPRFC(1) KGAFRLAKVLIRHY(1) RHVPKANKGADRSC(1) KTSWVRAAALVVVH(1) KSVNKDVRISLRD(1) KCIARRGRLPVKRY(1) KVLFRHARSSCKHY(1) KVL 46 Ribosome display (common panning) PMID:16878349 Model membranes Not determined. Not determined. Not determined. ZXXXZXXZXXXZXX ribosome display library Following stabilization of the peptide-ribosome-mRNA complex by addition of 50 mM Mg(2+), it was incubated with the model membrane immobilized on magnetic beads via streptavidin. mRNA was eluted from the bound complex in buffer containing EDTA. Of the resultant colonies, 50 clones were subjected to sequencing, of which 47 encoded a full-length peptide without a deletion. Sequences such as KVZ and RVZ were present in several of the peptides, and K/R was often followed by a hydrophobic amino acid in most of these, and rarely by a hydrophilic but nonpolar amino acid, but never by a basic or acidic amino acid. 1682 HHCRGHTVHSHHRCIG(6)[94] HHCRGHTVHSHHHCIR(2)[70] HHCRGHTVHSHHRCII(2)[NT] 3 Ribosome display (subtractive panning) 6 PMID:18613123 Cobalt(II) complex, Co(II) Not determined. Not determined. Not determined. X(2)-C-X(10)-C-X(2) ribosome display library ELISA,Surface plasmon resonance (SPR) SPR measurements were performed on a Biacore 2000 instrument with CM5 sensor chip. The dissociation constants KD (μM) were shown. NT denotes not tested. In vitro selection was carried out for the cobalt(II) complex immobilized on resin (Co-IR). The ribosomal conjugates displaying peptides with specific affinities to Co-IR were eluted and dissociated by adding imidazole and EDTA solutions to recover the corresponding mRNAs. Before the fourth, fifth, and sixth rounds of selection, the translation solution was incubated with bare resins for 40 min to exclude mRNAs of peptides that bound nonspecifically to bare resins. After six rounds of selection, 20 clones were chosen from the selected library and the sequences were analyzed. Table shows the peptide sequences that were found in more than two clones. The sequences were histidine-rich and similar to each other. In the major group of peptides, six histidines were found in 14 amino acid sequences except for the cysteines for cyclization. 1683 DYKGLDRGVI DYKFEDNLEC DYKFLDEVQN DYKWEDKVCI ARILDCKMDDD DYKMMDRRDF DYKCHDTGTD ALRSDYKWTD VDYQLRGAGY DYKSTDKWGY DYKPSVNEGG AYAIDYKRTD ISRDYKVDDA GDYKFADDMA TSHWDYKCWE SYKDFDLRKG DYKFMGYVAS FDDYKMTDIY D-Y-K-x(2)-D 18 Ribosome display (competitive panning) 2 PMID:19228777 Anti-FLAG M2 monoclonal antibody FLAG fusion protein Not determined. Not determined. X10 ribosome display library Dynabeads M-280 Streptavidin (Invitrogen, Carlsbad, CA, USA) was used for the immobilization of biotinylated anti-FLAG M2 antibody. To eliminate non-specific ternary complexes, the library was then incubated with 50ml magnetic beads without ligands for 30 min at room temperature. Remained supernatant was incubated with monoclonal antibody-immobilized magnetic beads for 1 h at room temperature. Elution of the ternary complex was carried out by the addition of Wash buffer with 100 mg/ml FLAG peptide in the case of competitive elution. The library after the second round of selection was cloned and sequenced. Out of 19 randomly picked clones, 12 clones possessed the consensus epitope equence for the anti-FLAG M2 antibody (DYKXXD), six possessed a partial epitope sequence and only one did not possess any consensus sequence. 1684 STKFDSVFGV(12) YLDSCCRSPW(9) STKFDALCSG(3) 3 Ribosome display (competitive panning) 3 PMID:19228777 Anti-β-cat monoclonal antibody 36a Catenin beta-1 Not determined. Not determined. X10 ribosome display library Dynabeads Protein G (Invitrogen, Carlsbad, CA, USA) was used for the immobilization of anti-b-Cat mAbs. To eliminate non-specific ternary complexes, the library was then incubated with 50ml magnetic beads without ligands for 30 min at room temperature. Remained supernatant was incubated with monoclonal antibody-immobilized magnetic beads for 1 h at room temperature. Elution of the ternary complex was carried out by the addition of Wash buffer with 20 nM β-Cat in the case of competitive elution. Irrespective of the selection round, obtained sequences contained putative epitope motifs that were found in the sequence of the original antigen, β-Cat. STKFD sequence, which has a homology for the β-Cat(111-115) (STQFD; amino acid residues 111-115 of β-Cat), was observed in the selected peptides. The YLDS sequence that is identical to the β-Cat(29-32) (YLDS; amino acid residues 29-32 of β-Cat) was also found. 1685 TLVMDLNAIE(7) TLVVGLDAID(1) 2 Ribosome display (competitive panning) 3 PMID:19228777 Anti-β-cat monoclonal antibody 48a Catenin beta-1 Not determined. Not determined. X10 ribosome display library Dynabeads Protein G (Invitrogen, Carlsbad, CA, USA) was used for the immobilization of anti-b-Cat mAbs. To eliminate non-specific ternary complexes, the library was then incubated with 50ml magnetic beads without ligands for 30 min at room temperature. Remained supernatant was incubated with monoclonal antibody-immobilized magnetic beads for 1 h at room temperature. Elution of the ternary complex was carried out by the addition of Wash buffer with 20 nM β-Cat in the case of competitive elution. Irrespective of the selection round, obtained sequences contained putative epitope motifs that were found in the sequence of the original antigen, β-Cat. TLVMDLNAIE sequence and a related sequence (TLVVGLDAID), which have a homology for the β-Cat(3-15) (TQADLMELDMAME; amino acid residues 3-15 of β-Cat), were found to be enriched. 1686 GFGRYRRHGSPW(1)[50] MAIGPYPACGSG(1)[43] LSVLVISMFNAV(1)[NB] MARHRNWPLVMV(1)[19] 4 Ribosome display (subtractive panning) 12 PMID:20479194 Envelope glycoprotein E2 Not determined. Not determined. Not determined. X12 ribosome display library Surface plasmon resonance (SPR) SPR measurements were performed using a Biacore 1000 biosensor. The dissociation constants KD (nM) were shown. NB denotes no binding. E2-GST-Sepharose 4B was utilized as a selection target. GST-Sepharose 4B was utilized as a counterselection target. Thirty individual clones were selected for DNA sequencing and then translated into 12-mer amino acid sequences. The results of an NCBI BLAST analysis showed that the four selected 12-mer peptides have protein-binding potentials. The selected peptide MARHRNWPLVMV demonstrated the highest specificity and affinity to the HCV E2 protein. Furthermore, amino acids 489 to 508 (YPPRPCGIVPAKSVCGPVYC) of E2 were identified as crucial for binding to PE2D. The selected peptides, especially MARHRNWPLVMV, not only dramatically blocked E2 protein binding to hepatocytes but also dramatically inhibited HCV cell culture (HCVcc) entry into hepatocytes. HCVcc and HCV particles from HCV patient serum samples could also be specifically captured using peptide MARHRNWPLVMV. 1687 RSDRGKAHPSRRS(1) RSSGYMTDTGPRSKHHPRNRETRS(1) RSVHASHHHIMRS(1) RSRLGHPVNHHRSRLAEQISQRRS(1) RSHARAERHHQRS(1) RSESRVDPAADRSHARAERHHQRS(1) RSFSVTSYSNGRSVGQVEPAGRRSQPPDVTRPRRS(1) RSVLRHLHLRRRSGDDLHETAARS(1) RSKLGHSPTIHRSEDQSAQAHARS(1) RSLQLRTGPGLRSHPIGRRTKHRS(1) RSGVHRNRIHKRS(1) RSHELHTHARSRS(1) RSQLARHHKHIRS(1) RSTGIHVIHHMRS(1) RSMNPRTHATTRSNMHHAAGASRS(1) 15 Bacterial display (common panning) 3 PMID:11722894 Zinc ion, Zn(2+) Not determined. Not determined. Not determined. X9 E. coli bacterial display library Bacterial cells were bound to zinc ions by use of stripped Ni(2+)-nitrilotriacetic acid (NTA) solid matrix (Qiagen) recoated with Zn(2+) by a standard method. Of the 20 clones, 15 displayed a Zn(2+)-binding phenotype. None of the isolated sequences showed similarity to known Zn(2+)-binding proteins, indicating that completely novel Zn(2+)-binding peptide sequences had been isolated. By changing the protein scaffold system, we demonstrated that the Zn(2+)-binding seems to be uniquely mediated by the peptide insert and to be independent of the sequence of the carrier protein. 1688 EWACNDRGFNCQLQR(1)[1] FPIYNQRGFITLASP(1)[3] MIFNSRGFLSLMSSG(1)[8] YPPRFQYYRFYYRGP(1)[5] HMRWNTRGFLYPAMS(1)[NT] RYIMNHRGFYIFVPR(1)[NT] VRTWNDRGFQQSVDR(1)[NT] LMNWRGFMVPRESPK(1)[NT] WTKLKNSRGFELQLD(1)[NT] PYLNARGFSVTREQI(1)[NT] TDFLSYYRVYRTPLQ(1)[NT] TFMPSYYRSWGPPPT(1)[NT] TTCKYYLSCRWRKDL(1)[NT] I-x-N-x-R-G-F, SYYRSY 13 Bacterial display (subtractive panning) PMID:15531628 C-reactive protein Not determined. Not determined. Not determined. X15 E. coli bacterial display library Flow Cytometry The apparent binding affinities of a subset of the selected peptides were determined using flow cytometric analysis. KDs (nM) measured by flow cytometry were shown. NT denotes not tested. For negative selection, 150ml of streptavidin-coated magnetic beads (QIAgen) were incubated with cells. For positive selection, biotinylated antigen (typically 1-100 nM) was added to the supernatant fraction and incubated on ice for 30-60 min. Cells were centrifuged as above and resuspended in 7.5 ml of cold PBS with 150ml of streptavidin-coated magnetic particles (QIAgen or Miltenyi). Consensus sequences were readily apparent for the target protein after two to three rounds of magnetic selection and one or two rounds of FACS. 1689 RLEICQNVCYYLGTL(1)[10] ICSYVMYTTCFLRVY(1)[8] TVLICMNICWTGETQ(1)[4] VTSLCMNVCYSLTTY(1)[NT] YWVCMNVCMYYTARQ(1)[NT] LPVWCVMHVCLTSSR(1)[NT] NEWYCQNVCERMPHS(1)[NT] IMMECFYVCTIANTQ(1)[NT] TWVQCTMVCYGMSTT(1)[NT] SITICWYTCMVQKTA(1)[NT] ADTICWYVCTISVHA(1)[NT] ICMNVC 11 Bacterial display (common panning) PMID:15531628 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. X15 E. coli bacterial display library Surface plasmon resonance (SPR),Flow Cytometry The apparent binding affinities of a subset of the selected peptides were determined using flow cytometric analysis. KDs (nM) measured by flow cytometry were shown. Besides, the YFP-SA-1(RLEICQNVCYYLGTL) insertional fusion exhibited an equilibrium dissociation constant of ~50 μM, as determined using surface plasmon resonance with 200 RU of immobilized streptavidin. NT denotes not tested. Consensus sequences were readily apparent for the target protein after two to three rounds of magnetic selection and one or two rounds of FACS. 1690 NPFCSWYRWRNWCTK(1) RHLYCWTWRWCHFKD(1) SYISTWLNFLFCGQS(1) NNYSAWLRCLLRAYS(1) C-x-W-x(2)-W-R-x-W, S-x-W-L-x(2)-L-x(4)-S 4 Bacterial display (common panning) PMID:15531628 Human serum albumin Not determined. Not determined. Not determined. X15 E. coli bacterial display library For negative selection, 150ml of streptavidin-coated magnetic beads (QIAgen) were incubated with cells. For positive selection, biotinylated antigen (typically 1-100 nM) was added to the supernatant fraction and incubated on ice for 30-60 min. Cells were centrifuged as above and resuspended in 7.5 ml of cold PBS with 150ml of streptavidin-coated magnetic particles (QIAgen or Miltenyi). Consensus sequences were readily apparent for the target protein after two to three rounds of magnetic selection and one or two rounds of FACS. 1691 GDTWVWYCWYWTRSI(1) WVCTWNYWTRVTWCL(1) PWCWMWTKGRWYYVA(1) QIQWCWVNHRWSPVV(1) WVAGYWWCWSVMYRS(1) TWTWCWRNYIWQLST(1) QEWRQLTRWCWVQIK(1) QTATVSYWCYWWWKV(1) W-V-x(4)-Y-W-T-R, W-C-W-x(3)-K 8 Bacterial display (common panning) PMID:15531628 Surface protein gp120, SU Not determined. Not determined. Not determined. X15 E. coli bacterial display library For negative selection, 150ml of streptavidin-coated magnetic beads (QIAgen) were incubated with cells. For positive selection, biotinylated antigen (typically 1-100 nM) was added to the supernatant fraction and incubated on ice for 30-60 min. Cells were centrifuged as above and resuspended in 7.5 ml of cold PBS with 150ml of streptavidin-coated magnetic particles (QIAgen or Miltenyi). Consensus sequences were readily apparent for the target protein after two to three rounds of magnetic selection and one or two rounds of FACS. 1692 QGRVMGPQQLYVMMR(1) NSLSMAPQQGWVQTG(1) TNKLMGPQQSKMFYW(1) GEGVMSPTQQRGPAR(1) AIVVSMTPFQQWSLS(1) AVHARHYTPWQQLYS(1) ERTGTYSPWQQLRVE(1) QQLSYGPQQQHAHMG(1) PMAIIGPQQMHLVLH(1) VYKSMAPQQTNAWQH(1) LQYSTAMGPQQMTSW(1) GSTSMGPQQFAWGTL(1) QKSTMTGWQQMGVMG(1) 13 Bacterial display (common panning) PMID:15531628 Anti-T7·Tag monoclonal antibody T7·Tag peptide MASMTGGQQMG Not determined. Not determined. X15 E. coli bacterial display library For negative selection, 150ml of streptavidin-coated magnetic beads (QIAgen) were incubated with cells. For positive selection, biotinylated antigen (typically 1-100 nM) was added to the supernatant fraction and incubated on ice for 30-60 min. Cells were centrifuged as above and resuspended in 7.5 ml of cold PBS with 150ml of streptavidin-coated magnetic particles (QIAgen or Miltenyi). Consensus sequences were readily apparent for the target protein after two to three rounds of magnetic selection and one or two rounds of FACS. 1693 AGFEG(1) SMWAHSSRDDAV(1) AKSSAKGKASGV(1) 3 Bacterial display (competitive panning) 5 PMID:16258189 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. FliTrx bacterial display library (X12) In-lab coated microtiter plates were blocked using bovine serum albumin or powdered skimmed milk in phosphate-buffered saline (PBS) buffer. Bound clones were eluted by displacing with excess biotin or mechanical shearing of flagella. Eight contained a stop codon in the region coding for the displayed random peptide. Given that the bacterial strain has no ability to suppress any stop codon, stop codon in this region is expected to stop the translation of functional flagellin. One of the bacterial clones selected expressed a pentapeptide instead of the expected dodecapeptide, most probably the result of an error in the process of library construction or a later mutation. 1694 YNCCHPMNNLCKE(1) EMCHPMWYVLCYH(1) SCCHPMSGVWCSM(1) KLCHPQAWYACTY(1) PSCHPQWYIVCWG(1) RSCHPMWWYLCES(1) FVCENVCYWVCDN(1) H-P-[QM] 7 Bacterial display (competitive panning) 4 PMID:16600968 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. OmpX7C bacterial display library ( X(2)-C-X(7)-C-X(2) ) Streptavidin binding peptides were enriched using two cycles of magnetic selection (MACS) with streptavidin-coated magnetic beads. Subsequently, the enriched population was screened using two cycles of FACS. In the final round of FACS, biotin was used as a competitor to favor the detection and sorting of clones with slow dissociation kinetics. 1695 QSLVCQNVCWMRE(1) CMIICQNVCYRKC(1) SKWICQNVCYPGL(1) KALVCQNVCYTMS(1) PTLICMNVCFYDQ(1) GTLVCMNFCYLSK(1) TRLICANLCWYAE(1) TYSWCANVCMHYS(1) SILRCENACYMVR(1) SHWFCVNVCFRIQ(1) LAWKCENVCYLEA(1) GAWSCWYVCERTT(1) L-[IV]-C-Q-N-V-C-Y 12 Bacterial display (competitive panning) 4 PMID:16600968 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. OmpX3C bacterial display library ( X(4)-C-X(3)-C-X(4) ) Streptavidin binding peptides were enriched using two cycles of magnetic selection (MACS) with streptavidin-coated magnetic beads. Subsequently, the enriched population was screened using two cycles of FACS. In the final round of FACS, biotin was used as a competitor to favor the detection and sorting of clones with slow dissociation kinetics. 1696 ERCWYVMHWPCNA(1) VNCGWMYYGPCTN(1) LRCHPQGGWSCIQ(1) H-P-[QM], C-G-W-M-Y-[FY]-x-E-C 3 Bacterial display (competitive panning) 4 PMID:16600968 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. CX7(1)C bacterial display library ( X(2)-C-X(7)-C-X(2) ) Streptavidin binding peptides were enriched using two cycles of magnetic selection (MACS) with streptavidin-coated magnetic beads. Subsequently, the enriched population was screened using two cycles of FACS. In the final round of FACS, biotin was used as a competitor to favor the detection and sorting of clones with slow dissociation kinetics. 1697 PKCGWMYYPECKV(1) LRCGWMYFEECKE(1) LICGWMYFEECDS(1) IYCGWMYYVECAY(1) MRCGWMYFPECLN(1) WVCHPMWEVMCLR(1) LVCHPQGPTWCIE(1) SWCHPQGMRECDW(1) VGCHPMAPLWCED(1) LWCHPQFLTTCHV(1) NQCHPQFQGWCGM(1) EWCHPQFNFLCGA(1) GTCWDHPQVGCNW(1) H-P-[QM], C-G-W-M-Y-[FY]-x-E-C 13 Bacterial display (competitive panning) 4 PMID:16600968 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. CX7(2)C bacterial display library ( X(2)-C-X(7)-C-X(2) ) Streptavidin binding peptides were enriched using two cycles of magnetic selection (MACS) with streptavidin-coated magnetic beads. Subsequently, the enriched population was screened using two cycles of FACS. In the final round of FACS, biotin was used as a competitor to favor the detection and sorting of clones with slow dissociation kinetics. 1698 VCRLMRGRCLLYSVF(1) TCVLHRQRCLMFTLR(2) ICVNIKKSLWACEIR(1) NCVRILMTFLDCTID(1) GCLQILPTLSECFGR(3) KQRGATMVLRTYTLR(1) YERRPTLVLRTWRPW(1) SVLVWVKDRGWRPAR(3) LYAHYDESRGWRWIR(1) WKFVWIINSRLREQA(1) WARVLLIEGRLIVCE(1) GNVLGKDYRLVKHVN(1) ARWIWYRNTATLNSV(1) CWILPYNTRTRCPLR(2) NQGLIGECHAYWCHG(2) NLIIGFCWLKKCPIR(1) LKVCGRYPGICDGIR(1) WWDMVSDRYIWKPVK(3) TQWIIPSKLAIKTPS(1) 19 Bacterial display (common panning) 3 PMID:16448666 Human breast ductal carcinoma cell line, ZR-75-1 Not determined. Not determined. Not determined. X15 E. coli bacterial display library Binding peptides were enriched after two rounds of co-sedimentation and one round of FACS. Of 70 isolated clones, 48 contained unique sequences. After screening, 36 of 48 of these exhibited binding to tumor cells as measured using flow cytometry. Within these 36 binding clones, phylogenetic analysis identified 19 clones sharing differing degrees of similarity. Isolated peptides could be categorized into several distinct groups possessing strong consensus sequences with as many as six identities. 1699 SSWCMRGQYNKICMW(2) 1 Bacterial display (subtractive panning) 3 PMID:16448666 Human breast ductal carcinoma cell line, ZR-75-1 Not determined. Not determined. Not determined. X15 CPX bacterial display library Binding peptides were enriched after two rounds of co-sedimentation and one round of FACS. Prior to the second round, a negative selection was performed by panning 5.0e7 bacteria against 1.0e7 adhered normal cells containing a mixture of immortalized human mammary cell line (MCF-10A) and Clonetics human mammary epithelial cells (HMEC). Bacteria not bound to normal cells in the supernatant were removed and immediately incubated with 1.0e7 suspended ZR-75-1 cells for a second round of positive selection using cosedimentation. Following an additional round of similar negative panning and positive coincubation with 10-fold fewer cells, tumor cells with fluorescent bacteria bound were immediately sorted using a FACSAria (Becton Dickinson, San Jose, CA). 1700 VPCQKRPGWVCLW(1) KWCVIWSKEGCLF(1) VECYLIRDNLCIY(1) WWCLGERVVRCAH(1) FYCVIERLGVCLY(1) RVCFLWQDGRCVF(1) 6 Bacterial display (subtractive panning) 3 PMID:16448666 Human breast ductal carcinoma cell line, ZR-75-1 Not determined. Not determined. Not determined. CX7(1)C bacterial display library ( X(2)-C-X(7)-C-X(2) ) Binding peptides were enriched after two rounds of co-sedimentation and one round of FACS. Prior to the second round, a negative selection was performed by panning 5.0e7 bacteria against 1.0e7 adhered normal cells containing a mixture of immortalized human mammary cell line (MCF-10A) and Clonetics human mammary epithelial cells (HMEC). Bacteria not bound to normal cells in the supernatant were removed and immediately incubated with 1.0e7 suspended ZR-75-1 cells for a second round of positive selection using cosedimentation. Following an additional round of similar negative panning and positive coincubation with 10-fold fewer cells, tumor cells with fluorescent bacteria bound were immediately sorted using a FACSAria (Becton Dickinson, San Jose, CA). 1701 RHTDGLRRIAAR(1) RTRRQGGDVSRD(1) RPRRSAARGSEG(1) ADRTRGRIRGNC(1) NTVWRLNSSCGM(1) EKWGMHQECYRH(1) TMEPRWWCNPIN(1) R-x-R-R 7 Bacterial display (common panning) 5 PMID:15236241 Cuprous oxide, Cu(2)O Not determined. Not determined. Not determined. FliTrx bacterial display library (X12) To recover binders, the surface was moved to a sterile culture dish containing 10 mL of IMC supplemented with carbenicillin and the plate was vortexed for 30 s to shear off the flagella. Peptides binding electrodeposited Cu(2)O with high avidity could be subdivided into two classes based on pI and hydrophilicity. In the hydrophilic and positively charged Class I binders, the Arg-X-X-Arg tetrapeptide appears to be implicated in metal oxide binding. Molecular dynamics simulations of the disulfide-constrained peptides suggest that the aforementioned motifs are important to properly orient two basic residues that are likely to contact the metal oxides. 1702 RIGHGRQIRKPL(1) VRTRDDARTHRK(1) PASRVEKNGVRR(1) MRHSSSGEPRLL(1) PAGLQVGFAVEV(1) RTDDGVAGRTWL(1) R-x(2)-R-K 6 Bacterial display (common panning) 5 PMID:15236241 Zinc oxide, ZnO Not determined. Not determined. Not determined. FliTrx bacterial display library (X12) To recover binders, the surface was moved to a sterile culture dish containing 10 mL of IMC supplemented with carbenicillin and the plate was vortexed for 30 s to shear off the flagella. Peptides binding electrodeposited ZnO with high avidity could be subdivided into two classes based on pI and hydrophilicity. In the hydrophilic and positively charged Class I binders, the Arg-X-X-Arg tetrapeptide appears to be implicated in metal oxide binding. Molecular dynamics simulations of the disulfide-constrained peptides suggest that the aforementioned motifs are important to properly orient two basic residues that are likely to contact the metal oxides. 1703 GMFTRNWCSRTFWCR(1) FIADPERYWCSVPAL(1) WYPVSWTKNLWYGPK(1) MCYYSRCNIMTCLSN(1) KYTWYGYSLRANWMR(1) RILCYAHARNRFIFN(1) 6 Bacterial display (common panning) PMID:17469847 Human red blood cells, RBCs Not determined. Not determined. Not determined. X15 E. coli bacterial display library To enrich for binding clones prior to screening by FACS, bacteria were incubated with human RBCs and washed to remove unbound bacteria. RBC-bound bacteria were then amplified by growth. Plasmid DNA was isolated from the first round pool and transformed into an E. coli MC1061 strain expressing green fluorescent protein (GFP), enabling subsequent rounds of screening using FACS. Since the bacteria used in the final two rounds of selection were fluorescent, RBCs were fluorescently labeled if bacteria were bound to the RBC membrane. RBCs binding to fluorescent bacteria were sorted by FACS, and the recovered bacteria were amplified by growth. Among isolated clones exhibiting binding to RBCs, six clones exhibiting the highest RBC labeling, as measured by flow cytometry, were selected for further analysis. Each of the isolated bacterial clones yielded greater than a 75-fold increase in RBC labeling relative to nonbinding cells displaying only the OmpA scaffold (i.e., without a peptide insertion). Clones GMFTRNWCSRTFWCR and KYTWYGYSLRANWMR bound most extensively to RBCs, resulting in more than 200-fold increased binding. 1704 GLVLETVGSGAR(1) CEAAQMVRPWVY(1) RSVSLSKSVAGV(1) GMEFGRRKRDVN(1) GARRHEIRGVDV(1) TDRSPRGVMRHA(1) 6 Bacterial display (common panning) PMID:18977188 Titanium dioxide, TiO2 Not determined. Not determined. Not determined. FliTrx bacterial display library (X12) Selection of proteins was carried out on a silicium wafer sputtered with TiO2 in anatase conformation. To verify binders and to analyze the binding kinetics of the diluted suspension of the purified proteins, the chip-based S-sens K5 surface acoustic wave sensor system was used. To recover binders, the surface was moved to a sterile culture dish containing 10 mL of IMC supplemented with carbenicillin and the plate was vortexed for 1 min to shear off the flagella. For peptide CGPGLVLETVGSGARGPC, the binding kinetics was analyzed. On- and off-rate binding constants were extracted from the fitted curves. With the resulting association rate constant k(on) and the dissociation constant k(off), the affinity of the peptide for the TiO2 surface was calculated, represented by the equilibrium dissociation constant K(D) = 81 nM. 1705 VRRSKHGARKDR[191] 1 Bacterial display (common panning) PMID:19048606 Heparin Not determined. Not determined. Not determined. FliTrx bacterial display library (X12) Surface plasmon resonance (SPR) The evaluation of the kinetic parameters was performed using a SPR-based BIAcoreX biosensor (BIAcore). The dissociation constant (KD =kd/ka, nm) is shown. Isolated insert analysis revealed a novel heparin-binding peptide sequence, VRRSKHGARKDR, designated as HBP12. Our analysis of the sequence alignment of heparin-binding motifs known as the Cardin-Weintraub consensus (BBXB, where B is a basic residue) indicates that the HBP12 peptide sequence contains two consecutive heparin-binding motifs (i.e. RRSK and RKDR). SPR-based BIAcore technology demonstrated that the HBP12 peptide binds to heparin with high affinity (K(D) = 191 nM). The HBP12 peptide is found to bind the cell surface HS expressed by osteoblastic MC3T3 cells and promote HS-dependent cell adhesion. Moreover, the surface-immobilized HBP12 peptide on titanium substrates shows significant increases in the osteoblastic MC3T3-E1 cell adhesion and proliferation. 1706 CHKRSFWADNC(1)[11.0, 10.9, 9.42] CRTQFRPNQTC(1)[10.1, 13.3, 13.9] CQLCDFWRTRC(1)[6.05, 8.72, 6.63] CFEDFNEQRTC(1)[1.68, 4.02, 3.50] CQNWIKDVHKC(1)[15.8, 19.4, 13.1] CLAKFLKGKDC(1)[NT] CWHRRTHKTFC(1)[NT] CRTIQTRSHWC(1)[NT] CIKLAQLHSVC(1)[NT] CWRHRNATEWC(1)[NT] 10 Bacterial display (common panning) PMID:20025916 IgG from human, mouse, rat, rabbit and goat Not determined. Not determined. Not determined. CX9C E. coli bacterial display library Surface plasmon resonance (SPR) The binding affinities of the selected peptides for human IgG, mouse IgG and rabbit IgG were determined using SPR analysis, respectively. SPR measurements were performed on a CM5 sensor chip. The binding constants (μM) were shown. FcBP1, DCAWHLGELVWCT, was used as positive control and its kinetic constants for human IgG, mouse IgG and rabbit IgG were 0.848, 0.732 and 1.69 μM, respectively. NT denotes not tested. The binding affinities of the selected peptides for IgGs were determined using SPR analysis. Five novel peptides were identified as potential affinity ligands for IgG purification. 1707 KMRAWGHPIWNW TKHGKRSRCYNL 2 Yeast display (common panning) 7 PMID:17238208 C crystalline face of synthetic sapphire, α-Al(2)O(3) Not determined. Not determined. Not determined. PL12 yeast display library (X12) Biopanned cells, each expressing a unique peptide, were agitated in the presence of sapphire surface, and then loosely bound cells were removed by surface washing. The adherent cells were then grown off the surface to create a subpopulation used in the subsequent round of biopanning. Twenty-four clones from sub-populations in the last round were sequenced. 1708 RTAKRKWKHTRD 1 Yeast display (common panning) 7 PMID:17238208 A crystalline face of synthetic sapphire, α-Al(2)O(3) Not determined. Not determined. Not determined. PL12 yeast display library (X12) Biopanned cells, each expressing a unique peptide, were agitated in the presence of sapphire surface, and then loosely bound cells were removed by surface washing. The adherent cells were then grown off the surface to create a subpopulation used in the subsequent round of biopanning. Twenty-four clones from sub-populations in the last round were sequenced. 1709 RTAKRKWKHTRD KRHKQKTSRMGK KRSKKCLRKNGS 3 Yeast display (common panning) 7 PMID:17238208 R crystalline face of synthetic sapphire, α-Al(2)O(3) Not determined. Not determined. Not determined. PL12 yeast display library (X12) Biopanned cells, each expressing a unique peptide, were agitated in the presence of sapphire surface, and then loosely bound cells were removed by surface washing. The adherent cells were then grown off the surface to create a subpopulation used in the subsequent round of biopanning. Twenty-four clones from sub-populations in the last round were sequenced. 1710 MEMNKVWRDLAA(1) IDQDKFWRELGS(1) LEGDKVWLEVRS(1) IEVDKVQHDLLS(1) FDEHKLWYELAA(1) LDVDKFREEVAS(1) 6 Yeast display (common panning) 2 PMID:19863063 Lipoate-protein ligase A Not determined. Not determined. Not determined. LAP yeast display library 1711 LERNKVWYEIKA(1) IEVDKSWLELRS(1) MELDKAWVEVWS(1) IDIDKIWYEFGS(1) FENDKIWHDIWA(1) IQGDKIWTELDS(1) FEYDKVWVDLPA(1) 7 Yeast display (common panning) 3 PMID:19863063 Lipoate-protein ligase A Not determined. Not determined. Not determined. LAP yeast display library 1712 FELDKVWFDVDS(1) FEIDKVWHDFPA(2) FEHEKVWYDLCA(5) FEINKVWFELLA(3) 4 Yeast display (common panning) 4 PMID:19863063 Lipoate-protein ligase A Not determined. Not determined. Not determined. LAP yeast display library The clones were obtained from high phycoerthyrin gate (Gate A). 1713 VEHDKVFYEFDS(1) IEIDKVWHDLYS(1) LEIDKVWHELDS(1) IELYKVWYEIDA(1) LEEDKIWYEFEA(1) VERDKVWYDISS(1) MERAKVWYELEA(1) 7 Yeast display (common panning) 4 PMID:19863063 Lipoate-protein ligase A Not determined. Not determined. Not determined. LAP yeast display library The clones were obtained from low phycoerthyrin gate (Gate B). 1714 YPSAPPQWLTNT(16) STPLVTGTNNLM(8) QSGSHVTGDLRL(2) ATTLHPPRTSLP(1) WPYAASNALVSP(3) AEMAAPTGLQVK(1) HLPTSSLFDTTH(1) 7 Phage display (subtractive panning) 5 DOI:10.1039/B806797J Titanium dioxide, TiO2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Peptide-bearing phage particles that bind to silica were first removed from the library by incubation with the diatom silica. The fourth and fifth rounds of such biopanning were conducted using 0.8% TBST washing solutions, in order to increase the stringency of selection for titania-binding phage. While prior phage display biopanning with silica and titania targets has led to the isolation of polycationic peptides enriched in basic residues, the subtractive biopanning process yielded several acidic peptides enriched in hydroxyl-bearing residues. These peptides were found to induce the precipitation of titania, but not silica, from aqueous precursor solutions at pH 3-8. 1715 CNKHQPMHC(1) CQNPMQTFC(1) CNQLSTRPC(1) CNNKVPVLC(1) CLQNRQSQC(1) CQLQRQWNC(1) CQVNSAHQC(1) 7 Phage display (common panning) 3 DOI:10.1039/B307593A Zinc sufide, ZnS Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Isolated ZnS binding motifs contained arrangements of positively charged groups (R, H, K), proline (a structural amino acid), and methionine residues. 1716 CQSMPHNRC(1) CFPMRSNQC(1) CPPQPNRQC(1) CNHQMPMQC(1) CNRQAVNAC(1) 5 Phage display (common panning) 4 DOI:10.1039/B307593A Zinc sufide, ZnS Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Isolated ZnS binding motifs contained arrangements of positively charged groups (R, H, K), proline (a structural amino acid), and methionine residues. 1717 CHMAPRWQC(1) CQSMPHNRC(2) CNNPMHQNC(3) 3 Phage display (common panning) 5 DOI:10.1039/B307593A Zinc sufide, ZnS Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Isolated ZnS binding motifs contained arrangements of positively charged groups (R, H, K), proline (a structural amino acid), and methionine residues. 1718 RRQDVHLPSRTL LRRSSEAHNSIV LPRAFMGHAPGS TRHMASRTEAHL LPPAWAMQVHTA PRPSPKMGVSVS 6 Phage display (common panning) DOI:10.1039/B307593A Zinc sufide, ZnS Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The peptide sequence typically had a combination of a positively charged amino acid group and a proline, followed by either a methionine, or a hydroxyl containing amino acid group. 1719 QNPIHTH 1 Phage display (common panning) DOI:10.1039/B307593A Lead sulphide, PbS Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 1720 CTYSRLHLC 1 Phage display (common panning) DOI:10.1039/B307593A Cadmium sulfide, CdS Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 1721 PWIPTPRPTFTG SLTPLTTSHLRS 2 Phage display (common panning) DOI:10.1039/B307593A Cadmium sulfide, CdS Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1722 SCFWFLRWSLFIVLFTCCS(1) SCESVDCFADSRMAKVSMS(1) SCVGFFCITGSDVASVNSS(1) SCSDCLKSVDFIPSSLASS(1) SCAFDCPSSVARSPGEWSS(1) SCMLFSSVFDCGMLISDLS(1) SCVDYVMHADSPGPDGLNS(1) SCSENFMFNMYGTGVCTES(1) SCSSFEVSEMFTCAVSSYS(1) SCGLNFPLCSFVDFAQDAS(1) 10 Phage display (common panning) 3 DOI:10.1002/adma.200700029 6Al-4V Ti Not determined. Not determined. Not determined. SC-X(16)-C M13 phage display library 1723 KSLSRHDHIHHH TQHLSHPRYATK SRPSRQPSASPT LLADTTHHRPWT 4 Phage display (common panning) 5 DOI:10.1021/cm071515t (001)-oriented TiO2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A 0.2% TBST wash solution was used to remove phages with low binding affinity to titania during the first four rounds, whereas a more stringent 0.5% TBST wash solution was used for the fifth round. 1724 MRMIRRFPSSLK TQHLSHPRYATK LKMNPSISSSLK FAVNPSKPAYFK FTTSNHTSRHGS YPMLPQNKHAQF QNLINWPPPRFS NVTTMTNHLVYS LATPFTATSATG 9 Phage display (common panning) 5 DOI:10.1021/cm071515t (100)-oriented TiO2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A 0.2% TBST wash solution was used to remove phages with low binding affinity to titania during the first four rounds, whereas a more stringent 0.5% TBST wash solution was used for the fifth round. 1725 RKKRTKNPTHKL KSLSRHDHIHHH TQHLSHPRYATK ASIEELRVPRQA NHHHQPLARNQS LLPQNGSTPRHS SFSAITKNVHWM TFSNPLYMWPRP SPGLSLVSHMQT SAHGTSTGVPWP 10 Phage display (common panning) 5 DOI:10.1021/cm071515t (110)-oriented TiO2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A 0.2% TBST wash solution was used to remove phages with low binding affinity to titania during the first four rounds, whereas a more stringent 0.5% TBST wash solution was used for the fifth round. 1726 TTAAVDMPRSTP(3) HESFWYLPHQSY(1) LLADTTHHRPWT(8) QSNYPRASYVFQ(1) KSLSRHDHIHHH(2) MPWAHRAPQGIA(1) 6 Phage display (common panning) 6 DOI:10.1021/cm071515t (001)-oriented TiO2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A 0.2% TBST wash solution was used to remove phages with low binding affinity to titania during the first four rounds, whereas a more stringent 0.5% TBST wash solution was used for the fifth round. A additional round of screening were conducted with a 0.5% TBST wash solution (for a total of six rounds of biopanning). 1727 LLADTTHHRPWT(2) SHAFTASPRYLH(4) KSLSRHDHIHHH(6) YAKSPPTPYYTP(4) LAPKPFEPRYTR(1) 5 Phage display (common panning) 7 DOI:10.1021/cm071515t (001)-oriented TiO2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A 0.2% TBST wash solution was used to remove phages with low binding affinity to titania during the first four rounds, whereas a more stringent 0.5% TBST wash solution was used for the fifth round. Two additional rounds of screening were conducted with a 0.5% TBST wash solution (for a total of seven rounds of biopanning). 1728 KSLSRHDHIHHH(17) SRPSRQPSASPT(3) 2 Phage display (common panning) 8 DOI:10.1021/cm071515t (001)-oriented TiO2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A 0.2% TBST wash solution was used to remove phages with low binding affinity to titania during the first four rounds, whereas a more stringent 0.5% TBST wash solution was used for the fifth round. Three additional rounds of screening were conducted with a 0.5% TBST wash solution (for a total of eight rounds of biopanning). 1729 DPIYALSWSGMA(1) TMATYVNQSLTG(1) SVSLPYANLATH(1) SLYNTAASHVPT(1) DLNTNRTGMVLH(2) NYLHNHPYGTVG(1) LTPTSRPTPYPA(1) KSLSRHDHIHHH(9) NFRPVTAMPRLD(1) NINANAAQIKRH(1) 10 Phage display (common panning) 6 DOI:10.1021/cm071515t (001)-oriented TiO2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A 0.2% TBST wash solution was used to remove phages with low binding affinity to titania during the first four rounds, whereas a more stringent 0.5% TBST wash solution was used for the fifth round. A additional round of screening were conducted with a 0.8% TBST wash solution (for a total of six rounds of biopanning). 1730 TQHLSHPRYATK(9) YAKSPPTPYYTP(8) KSLSRHDHIHHH(5) 3 Phage display (common panning) 7 DOI:10.1021/cm071515t (001)-oriented TiO2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A 0.2% TBST wash solution was used to remove phages with low binding affinity to titania during the first four rounds, whereas a more stringent 0.5% TBST wash solution was used for the fifth round. Two additional rounds of screening were conducted with a 0.8% TBST wash solution (for a total of six rounds of biopanning). 1731 APPGHHHWHIHH MSASSYASFSWS KPSHHHHHTGAN 3 Phage display (common panning) 3 PMID:12908327 Silica, Si Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1732 MSPHPHPRHHHT MSPHHMHHSHGH LPHHHHLHTKLP APHHHHPHHLSR RGRRRRLSCRLL 5 Phage display (common panning) 4 PMID:12908327 Silica, Si Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1733 LKAHLPPSRLPS 1 Phage display (common panning) PMID:16601154 Gold Not determined. Not determined. Not determined. X12 M13 phage display library 1734 RKLPDAPGMHTW(34) LDTTNVSGPMSS(1) SYRLPVYLHALL(1) SDPNQDWRRTTP(1) LPSQLLSQVNLT(1) LCANNTTSVHPP(1) MQMEGKPTLTLR(1) STLKNPINLLAN(2) QDMIRTSALMLQ(1) SCHVWYDSCSSP(1) 10 Phage display (common panning) 3 PMID:14624545 Titanium, Ti Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After three rounds of panning procedures carried out against Ti particles, we observed that 33 of 43 phages displayed on their surfaces peptides having the identical sequence. 1735 NFMSLPRLGHMH TSNAVHPTLRHL 2 Phage display (common panning) PMID:19422199 Pd wire Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Both peptide sequences possessed basic pI values; however, the peptide TSNAVHPTLRHL sequence possessed more basic and hydroxyl-containing residues, comparatively. Additionally, theoretical modeling of the peptide Pd surface binding capabilities suggests that the two histidine residues bind to the materials surface in a pinched arrangement, which will likely lead to open interaction sites between the solution and the metallic surface. 1736 TGTSVLIATPYV 1 Phage display (common panning) PMID:20083240 Gold Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Metallic gold powder in an Eppendorf tube was directly used as the target material for isolation of binding phages instead of a substrate coated with the target material or a solution of phage with a target attached by affinity tags. 1737 LNAAVPFTMAGS 1 Phage display (common panning) 4 PMID:20627315 Titanium dioxide, TiO2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) First, three rounds of phage display selection were performed in an Eppendorf PP tube containing a 5-mm-diameter and 2 mm amorphous-TiO2-coated disk. Secondly, an additional round of selection was performed on newly shaped surfaces. The TiO2-binding peptide sequence (LNAAVPFTMAGS) occurred in 50% of the clones. 1738 MTWDPSLASPRS 1 Phage display (common panning) 4 PMID:20627315 Stainless steel Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) First, three rounds of phage display selection were performed in an Eppendorf PP tube containing a 5-mm-diameter and 2 mm thick stainless steel. Secondly, an additional round of selection was performed on newly shaped surfaces. The stainless steel-binding peptide sequence (MTWDPSLASPRS) occurred in 70% of the clones. 1739 YQLRPNAESLRF 1 Phage display (common panning) 4 PMID:20627315 Bulk steel (edges) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A dominant peptide sequence for recognition of the steel of the edges was highlighted (YQLRPNAESLRF). 1740 AGETQQAM 1 Phage display (common panning) 3 PMID:20383127 Iridium oxide, IrO2 Not determined. Not determined. Not determined. X8 M13 phage display library 1741 MFLRAG LHADVW QLYMKG WAMSEV EPGLDR SFNILR PVQASP DSASHS SPCRGL GWWPHR 10 Phage display (subtractive panning) 3 PMID:7892246 Capsid protein Not determined. Not determined. Not determined. f3-6mer phage display library (X6) The library had been incubated with a buffer containing BSA, before panning, to eliminate phage that bind BSA. The phage library was screened against truncated HBcAg (aa 3-148). The washing buffer used during affinity purification contained 0.15M NaCl. 1742 LLGRMK(15)[0.17 ± 0.01, 0.22 ± 0.01] YLLRFR(11)[1.53 ± 0.10, ND] LLGRLK(6)[1.13 ± 0.05, ND] LLGRFK(2)[0.64 ± 0.03, ND] LLGRFR(2)[NT] LLGRLR(1)[NT] LLGRMR(1)[NT] SLWKWK(1)[NT] WTFLRG(1)[NT] L-L-G-R-[MFL]-[KR] 9 Phage display (subtractive panning) 3 PMID:7892246 Capsid protein Not determined. Not determined. Not determined. f3-6mer phage display library (X6) Binding assay Relative dissociation constants (KdRel, μM) of the phage-truncated HBcAg complexes and the phage-full-length HBcAg complexes were shown, respectively. ND represents not determined. NT denotes not tested. The library had been incubated with a buffer containing BSA, before panning, to eliminate phage that bind BSA. The phage library was screened against truncated HBcAg (aa 3-148). The washing buffer used during affinity purification contained 0.5M NaCl. In the first round of panning, 0.001% of phage bound to the membrane and this binding increased to 1% in the third panning. Of 46 independent clones, the sequence LLGRMK predominated. Six sequences are considered unrelated and not given in the original paper. LLGRMK-bearing phage bound to full-length HBcAg (aa 3-183) as efficiently as to truncated HBcAg but did not bind to heat-denatured, truncated HBcAg. The sequence LLGRMK could represent one loop of the binding site of an antibody to HBcAg, but more interestingly it may resemble a region of the HBsAg polypeptide that contacts the nucleocapsid in the intact virion. 1743 ANGCCD HAILNI ESQSVL RVADIV SKGTVA DVSIGV RYLVLE NINSPV IWLSNG WPRGAG 10 Phage display (subtractive panning) 3 PMID:7892246 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. f3-6mer phage display library (X6) The library had been incubated with a buffer containing BSA, before panning, to eliminate phage that bind BSA. Thus, the peptides here are considered to be random sequences. 1744 CFDYAPYVSAVDDIC LSAVDDLATVVYGSR AGCTALSAVDDVPGC ASSAVDDAPWAVITF VGLLSVEELVPGGAA GSFSAEHFLDDFAIW RNVPPIFNDVYWLAF GGVSAVPFMDYFPSW [ST]-A-V-D-D, S-A-V-P-x(2)-D, S-A-x(4)-D 8 Phage display (common panning) 2-4 PMID:10456916 Anti-RESA monoclonal antibody MAb 18/2 Ring-infected erythrocyte surface antigen Not determined. Not determined. f3-15mer phage display library (X15) Most phage isolated from this library bound to MAb 18/2-coated ELISA plates. The binding of representative peptides, CFDYAPYVSAVDDIC and GSFSAEHFLDDFAIW, to MAb 18/2 has been confirmed. 1745 QTAVDDAEAAAKCRHCI QDETRSAVDSIPQAHCS GLKNCTVQPWDATDVCD QISAVS LPAQPNACWPTMTPLCA YVGSQSEDRDMSCGHCS QQNDLLVYSAVPLSDCK ASAAEGDD [ST]-A-V-D-D, S-A-V-P-x(2)-D, S-A-x(4)-D 8 Phage display (common panning) 2-4 PMID:10456916 Anti-RESA monoclonal antibody MAb 18/2 Ring-infected erythrocyte surface antigen Not determined. Not determined. X15CX phage display library Interestingly, sequences of two peptides QISAVS and ASAAEGDD align with the second motif only if the phage protein backbone of gpVIII to which the peptides are fused is considered to contribute to the binding. Most phage isolated from this library bound to MAb 18/2-coated ELISA plates. In contrast, phage lacking a peptide (wild-type M13 phage) did not bind to MAb 18/2, and one phage clone with the insert sequence YVGSQSEDRDMSCGHCS, which lacks the consensus motif, also did not bind. RESA protein inhibits binding of MAb 18/2 to selected phage. The binding of a representative peptide, GLKNCTVQPWDATDVCD, to MAb 18/2 has been confirmed. 1746 LWVGGGRNA(6) 1 Phage display (common panning) 3 PMID:10867191 Grass carp hemorrhage virus Not determined. Not determined. Not determined. X9 fUSE5 phage display library To remove weakly or nonspecifically bound peptidea??a Sixteen clones which inhibited the replication of GCHV in a grass carp kidney cell line were selected. The TCID 50 of GCHV was decreased over 10 000×. Six clones having the strongest inhibitory effect shared the same DNA sequence, with a deduced amino acid sequence of NH(2)-Leu-Trp-Val-Gly-Gly-Gly-Arg-Asn-Ala-COOH. 1747 CTLTTKLYC CQLGGPSHC CQKGGPSHC CQHEMPSKC 4 Phage display (common panning) 1 PMID:12021868 Newcastle disease virus, NDV Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A fusion phage carrying the amino acid sequence TLTTKLY was selected from the panning procedure. An antibody competition assay showed that the selected phage was capable of competing with the polyclonal antibodies raised against NDV for binding sites on the virus. Determination of the binding affinity of this phage with NDV by an equilibrium binding assay in solution revealed two different dissociation constants, suggesting that there could be two distinct binding sites for the phage on NDV. Synthetic peptides with the sequence CTLTTKLYC, either in linear or cyclic conformations inhibited the binding of phage bearing the same sequence to NDV. 1748 CTLTTKLYC CTLTTKAYC CTLTTPELC CTPTPRLYC CTPSPRLYC CTVWTSEHC CQLGGPSHC CQKGGPSDC CQKGGPSHC 9 Phage display (common panning) 2 PMID:12021868 Newcastle disease virus, NDV Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A fusion phage carrying the amino acid sequence TLTTKLY was selected from the panning procedure. An antibody competition assay showed that the selected phage was capable of competing with the polyclonal antibodies raised against NDV for binding sites on the virus. Determination of the binding affinity of this phage with NDV by an equilibrium binding assay in solution revealed two different dissociation constants, suggesting that there could be two distinct binding sites for the phage on NDV. Synthetic peptides with the sequence CTLTTKLYC, either in linear or cyclic conformations inhibited the binding of phage bearing the same sequence to NDV. 1749 CTLTTKLYC CNLTTKLYC 2 Phage display (common panning) 3 PMID:12021868 Newcastle disease virus, NDV Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Thirty-nine out of 40 phages analyzed from the third round carried the sequence TLTTKLY and only one related sequence was identified in which its first residue, Thr (T), was substituted by Asn (N). An antibody competition assay showed that the selected phage was capable of competing with the polyclonal antibodies raised against NDV for binding sites on the virus. Determination of the binding affinity of this phage with NDV by an equilibrium binding assay in solution revealed two different dissociation constants, suggesting that there could be two distinct binding sites for the phage on NDV. Synthetic peptides with the sequence CTLTTKLYC, either in linear or cyclic conformations inhibited the binding of phage bearing the same sequence to NDV. 1750 CHPQFEALC 1 Phage display (common panning) 3 PMID:12021868 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. Ph.D.-C7C phage display library (CX7C) Biopanning gave a consensus sequence of HPQFEAL, which was about 70% of the total phages screened from the third round of panning. 1751 GWRLLGFGPASSFSM TRLFRVPVLPSGVTS PFARAPVEHHDVVGL [AVLIFYW](2)-R-x-P-V-x(4)-V 3 Phage display (common panning) 4 PMID:12381731 Apical membrane antigen 1, AMA1 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) ELISA Phage clones displaying each of the three peptides bound to PfAMA1 in a dose-dependent manner, although the F1 (GWRLLGFGPASSFSM) and F3 (PFARAPVEHHDVVGL) peptides appeared to have an ~10-fold higher relative affinity compared with the F2 peptide (TRLFRVPVLPSGVTS). Absolute affinities were difficult to estimate from these data because the presence of up to five copies of peptide on each phage particle may impart avidity effects that are difficult to predict. Phage containing a peptide picked at random from the unpanned library and consisting of the sequence GDVWLFKTSTSHFAR (F5 peptide) were unable to bind to PfAMA1 even at phage concentrations of e11 colony-forming units/ml. Panning were performed on E. coli cell-expressed and refolded AMA1 from the 3D7 strain of P. falciparum. Three peptides with affinity for AMA1 were isolated, and characterization of their fine binding specificities indicated that they bind to a similar region on the surface of AMA1. Peptide GWRLLGFGPASSFSM was found to be a potent inhibitor of the invasion of P. falciparum merozoites into human erythrocytes. We propose that this peptide blocks interaction between AMA1 and a ligand on the erythrocyte surface that is involved in a critical step in malarial invasion. 1752 CFPWGNTWC CFPWGKEYC CFPWGNQWC CFPWGDQWC CFPWPLWAC CFPWGNEPC CFPWGDQCC CFPWGQTAC CFPWGDWPC FPWG 9 Phage display (common panning) 3 PMID:12951030 RNA-directed RNA polymerase NS5B Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Surface plasmon resonance (SPR) Peptides CFPWGKEYC, CFPWGNTWC, and CFPWGNQWC bound with varying affinities (Kd ranging from 33 to 56 μM) to the enzyme NS5B with a stoichiometry consistent with a 1:1 binding event. Surface plasmon resonance studies showed that three highly purified synthetic constrained peptides bound to immobilized NS5B with estimated Kd values ranging from 30 to 60 μM. In addition, these peptides inhibited the NS5B activity in vitro with IC 50 ranging from 6 to 48 μM, whereas in contrast they had no inhibitory effect on the enzymatic activities of calf thymus polymerase α, human polymerase β, RSV polymerase, and HIV reverse transcriptase in vitro. 1753 TWFNPFGYYSWA(5)[0.49] TWFWPYPYPHLP(3)[0.53] ENGLHNRSLNPR(3)[0.05] TLWPWAWRHNWQ(2)[0.43] TWWTGTYPWYPR(2)[0.48] TFWWHPNYYVDW(1)[0.15] GQPSHDPVPPTT(1)[0.05] SSTSTVTPAHST(1)[0.06] SSPLAHYLNAPT(1)[0.08] 9 Phage display (common panning) 3 PMID:14614029 Regulatory protein E2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA In phage ELISA, the HPV16 E2 protein was added to each well followed by addition of BSA blocking protein. BSA is immobilized separately in the absence of the E2 protein in the 96-well plate. The absorbance value (A) was determined by subtraction of the absorbance of BSA from the absorbance of the HPV16 E2. The raw A value of BSA was ~0.05 or 0.1. Among the isolated phage clones, authors found that tryptophan-rich peptide sequences appeared repetitively in successive cycles of phage library panning. Replacement of the tryptophan amino acids in these dodecapeptides reduced the degree to which these peptides bound to the E2 protein. These E2-binding peptides were tested for their ability to inhibit the transcriptional regulatory function of E2 in a test cell line, which contained an E2 gene and a luciferase reporter gene driven by an E2-dependent transcriptional promoter. 1754 CTGDARHRC(5) CNSVGRIWC(3) CTPATLLLC(2) CMRLGSSIC(2) CLVSFGLSC(2) CLDSSRGIC(2) CRQLGKQRC(1) CHQLGKQRC(1) CGGVHFAYC(1) CADVSQPVC(1) CAGFEKIPC(1) CNRLPRELC(1) CVLESHYEC(1) 13 Phage display (common panning) 4 PMID:15700752 Human antibodies eluted from the surface of infected red blood cell (iRBC) Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Of 23 randomly chosen clones that were sequenced, 13 individual sequences were detected at varying frequencies and 3 of the 13 sequences had homology with membrane proteins known to exist on iRBC. The majority of phage clones (7 out of 8 clones) selected after the 4th panning bound selectively to IgG in IHS. Specific binding of the selected phage to IgG in IHS was also confirmed using 24 IHS and 11 sera from uninfected individuals. One phage clone was the most frequently found in the sequenced clones after the 4th panning, and the binding of this clone to IgG in all IHS was greater than in any serum from uninfected individuals. 1755 CQSLPTHNC CSSQNGRIC CRTLPPILC CHLLLPRPC CQAMHNRFC CTPTVTRSC CTESTINTC CQYNPLPYC CNHTRNMAC CPSRLSSQC CTKYWARNC CHPTGLHQC CLTDRHRTC CTPSLPWLC CQGPVKPLC CDATSAQVC CKPTHYNSC CRHNNLHHC CEWTESMMC CSTNPPTQC CVNSPPTMC CTNPATPFC CLWPTLKGC CHTKIFPNC CHARNSTQC CSLTNTLLC CNPYNTSMC CNGVIHNQC CYHPSFNTC CSGSETKVC CYETKTHSC CTNRHAAGC CNLLRQQTC CVSGQESIC CLALNMSYC COPHTLPTC CNTKNFHSC CNPMKPLMC CPRSGTAYC CSTPELTFC CMTSHPTLC CPLGKLPWC CQSASPRLC CLTSPTSTC CNTSSTPHC CTQKNIAAC CTNSTLQSC CKPDAISVC CNQSTLLTC CHSPLTSSC CFPTTSRGC CGFSNFRSC CQSQNHNTC CQSIRGPMC CPSTGYSYC CPPGKSSMC CHNLKRPTC CLFNHPKYC CYSLLPRVC CTMLLFHRC CRNTDTALC CPPPQGQTC CIHAPHTQC CPGHIHRTC CLALRHSNC CSPQLAPFC CQLPAERWC CRSLTDNQC CLHKPWSRC CPFVKTQLC 70 Phage display (competitive panning) 6 PMID:15919886 Integrin alpha-V-beta-3, integrin αvβ3 Abciximab Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Abciximab was employed to elute phage from the plate-bound ??? Authors found that 11 peptides reduced the number of foci to a greater extent than did 80 μg/ml ReoPro when preincubated with Vero E6 cells. In addition, 8 of the 70 peptides had sequence similarity to SNV glycoproteins. Authors compared all 18 peptide sequences (10 most potent, 7 peptides with sequence similarity to hantavirus glycoproteins, and 1 peptide that was in the group that displayed the greatest potency and had significant sequence similarity) for their abilities to inhibit SNV, Hantaan virus (HTNV), and Prospect Hill virus (PHV) infection. There was a marked trend for the peptides to inhibit SNV and HTNV to a greater extent than they inhibited PHV, a finding that supports the contention that SNV and HTNV use β3 integrins and PHV uses a different receptor, β1 integrin. 1756 CETGAKPHC(0.2) CNPHPQQPC(0.2) CNPPPPQPC(0.2) CHPPHPQPC(0.2) CQHGAMGLC(0.2) 5 Phage display (common panning) 1 PMID:16087122 Large envelope protein Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Saturation equilibrium assay Results of equilibrium binding assay show that the concentration of the free phage clone CETGAKPHC reduces progressively with increased concentration of HBsAg. The binding data were fitted with the equation for two binding sites and the Krel D values were 2.9±0.9 nM (KrelD1) and 0.83±0.63 mM (KrelD2). The binding site of peptide ETGAKPH was located on the immunodominant region of HBsAg. An equilibrium binding assay in solution showed that the phage binds tightly to HBsAg with a relative dissociation constant (KDrel) of 2.9+/-0.9 nM. The phage bearing this peptide has the potential to be used as a diagnostic reagent and two assays for detecting HBsAg in blood samples are described. 1757 CETGAKPHC(0.5) CQTNHMGLC(0.2) CQTKHQGRC(0.1) CQPNHMGLC(0.1) CQSKPQGLC(0.1) 5 Phage display (common panning) 2 PMID:16087122 Large envelope protein Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Saturation equilibrium assay Results of equilibrium binding assay show that the concentration of the free phage clone CETGAKPHC reduces progressively with increased concentration of HBsAg. The binding data were fitted with the equation for two binding sites and the Krel D values were 2.9±0.9 nM (KrelD1) and 0.83±0.63 mM (KrelD2). The binding site of peptide ETGAKPH was located on the immunodominant region of HBsAg. An equilibrium binding assay in solution showed that the phage binds tightly to HBsAg with a relative dissociation constant (KDrel) of 2.9+/-0.9 nM. The phage bearing this peptide has the potential to be used as a diagnostic reagent and two assays for detecting HBsAg in blood samples are described. 1758 CETGAKPHC(0.75) CQTGEKPQC(0.083) CETGEKPQC(0.083) CQAFFPNAC(0.083) 4 Phage display (common panning) 3 PMID:16087122 Large envelope protein Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Saturation equilibrium assay Results of equilibrium binding assay show that the concentration of the free phage clone CETGAKPHC reduces progressively with increased concentration of HBsAg. The binding data were fitted with the equation for two binding sites and the Krel D values were 2.9±0.9 nM (KrelD1) and 0.83±0.63 mM (KrelD2). The binding site of peptide ETGAKPH was located on the immunodominant region of HBsAg. An equilibrium binding assay in solution showed that the phage binds tightly to HBsAg with a relative dissociation constant (KDrel) of 2.9+/-0.9 nM. The phage bearing this peptide has the potential to be used as a diagnostic reagent and two assays for detecting HBsAg in blood samples are described. 1759 IRPDTPR(1)[0.764 ± 0.043, 0.167 ± 0.047] KYLNAHV(1)[0.683 ± 0.015, 0.174 ± 0.023] NKVLVPP(1)[0.756 ± 0.011, 0.244 ± 0.042] VPLQPLR(1)[0.719 ± 0.038, 0.189 ± 0.013] KNVQVPL(1)[0.500 ± 0.043, 0.133 ± 0.030] KNVHPPP(1)[0.527 ± 0.017, 0.117 ± 0.030] SHQHARL(1)[0.578 ± 0.035, 0.129 ± 0.038] KNVHVPL(1)[0.572 ± 0.026, 0.123 ± 0.017] TPHTLPP(1)[0.353 ± 0.017, 0.095 ± 0.015] SLIQASA(1)[0.917 ± 0.022, 0.595 ± 0.017] 10 Phage display (competitive panning) 3 PMID:18430373 Interferon alpha/beta receptor 2, IFN-R-2 Interferon alpha-2 2HYM,2KZ1,2LAG,3S9D,3SE3, Not determined. Ph.D.-7 phage display library (X7) ELISA WISH cells prepared in 96-well plates were incubated with selected phage clones. Phage clones binding to WISH cells were detected by phage ELISA in the absence or presence of IFNα-2b, then the absorbance at 490 nm was analyzed. Wild-type M13 phage was used as a negative control. Data, expressed as mean of 3 independent experiments ± SD, were reproduced from Figure 2 in the reference. In the absence of IFNα-2b, the absorbance of M13 phage was 0.052 ± 0.033, while in the presence of IFNα-2b its value was 0.056 ± 0.017. The data in the first column of affinity values were obtained in the absence of IFNα-2b, the data in the second column of affinity values in the presence of IFNα-2b. Biopannings were performed with WISH cells, which endogenously express type I IFN receptors on the cell surface. Phages bounding to surviving cells were eluted with IFNα-2b. ts: Sixteen positive clones were obtained after 3 rounds of functional selection. Ten clones were picked from these positive clones according to the results of phage ELISA and were sequenced. The amino acid sequences homologous to IFNα-2b were defined by residues AB loop 31-37, BC loop 68-74, C helix 93-99, CD loop 106-112, D helix 115-121, DE loop 132-138, and E helix 143-161. Two of the peptides, designated clones T3 (IRPDTPR) and T9 (KNVHPPP), aligned with the IFNAR2-binding domains (AB loop and E helix), were synthesized and designated as IR-7 and KP-7, respectively. Both KP-7 and IR-7 were found to compete with GFP/IFNα-2b for receptor binding and mimicked the antiviral activity of IFNα-2b cooperatively. 1760 TSQNIRS(1) TSYHRSA(1) 2 Phage display (common panning) 3 PMID:21193836 Envelope glycoprotein E2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) When pep7-1 (TSQNIRS) was present, the infectivity of HCV particles in cell culture was notably decreased. This decrease was demonstrated by Western blot analysis, immunofluorescence assay, and reverse transcription PCR assay. 1761 SVSVGMKPSPRP(1) SVSVGTKPRPRP(1) SVSWGMKPSPRQ(1) 3 Phage display (common panning) 3 PMID:21193836 Envelope glycoprotein E2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Pep12-1 (SVSVGMKPSPRP) showed little inhibitory effect on HCV infection. 1762 CHRCFHFRRHPVAVF TRHRHVPRFLPLRHV 2 Phage display (common panning) 3 PMID:21154952 Interleukin-10, IL-10 Interleukin-10 receptor subunit alpha (IL-10RA) 1J7V,1Y6K, Not determined. f3-15mer phage display library (X15) Each selection was performed on the streptavidine-coated plates. Fifteen-mer peptides binding to IL-10 selected from the phage display library were synthesized and tested in a bioassay using the IL-10-sensitive MC/9 cell line to measure their IL-10 blocking activity. Peptides p9 (CHRCFHFRRHPVAVF) and p13 (TRHRHVPRFLPLRHV) inhibited human IL-10-induced proliferation. Peptide binding to IL-10 was demonstrated using surface plasmon resonance analysis. It was found that p9 and p13 bound to immobilized IL-10, as compared to a control peptide. 1763 DFRRLPGAFWQLRQP(13)[0.429, 24][0.2] GWWYKGRARPVSAVA(4)[0.398, 13][0.5] VWRLLAPPFSNRLLP(1)[0.488, 1][0.2] R-x-L-P-x(2)-F-x(3)-R-x-P 3 Phage display (common panning) 5 PMID:10456319 Ganglioside GM1 Cholera toxin B protein, CTB Not determined. Not determined. X15 fUSE5 phage display library ELISA,Quartz crystal microbalance (QCM) In phage ELISA, the absorbance at 492 nm was determined. Absorbances were reproduced from the graph when the concentration of phage was 5 nM. And the value of the control phage (displaying LGRAGQSYPSFARGL) was 0.221. Besides, in binding inhibition assay, horseradish peroxidase-conjugated CTB and synthetic peptides were incubated with the GM1 plates in a 24 well plate. The IC50 values (μM) of synthetic peptides were determined and shown. Furthermore the dissociation constants (Kd values, nM) were determined by QCM and shown. Three synthetic pentadecapeptides inhibited the binding of cholera toxin B subunit to the GM1 monolayer with an IC50 of 24, 13 and 1.0 μM, respectively. These peptides will be useful for searching functional roles of ganglioside GM1. 1764 ARLSPTMVHPNGAQP(2)[1.268 ± 0.179] GLAYSRFKVSRAIWR(2)[0.283] HRWMPHVFAVRQGAS(1)[0.319 ± 0.078] ALADSIWSSRSGPGL(1)[0.310 ± 0.136] KDFLSGLRHLHFLHS(1)[0.319 ± 0.122] TFRLPLVRAQYDSSH(1)[0.139] ARVMRQVCGGTGCGV(1)[0.270 ± 0.201] TSVNRGFLLQRASHP(1)[0.192] 8 Phage display (competitive panning) 4 PMID:20476787 Hemagglutinin, HA Sialyl Lewis(X), SLX Not determined. Not determined. X15 fUSE5 phage display library ELISA,Surface plasmon resonance (SPR) In phage ELISA, the absorbance at 492 nm was determined. Data shown were reproduced from the graph and expressed as means ± the standard deviation (n = 3). Besides, clone f1 (displaying ARLSPTMVHPNGAQP) was found to have a high affinity by SPR analysis, with dissociation constant (Kd) values of 7.8 and 8.6 nM against H1 and H3. A docking simulation suggested that, similar to sialic acid, the peptides are recognized by the receptor-binding site in HA, which indicates that these peptides mimic the sialic acid structure. N-stearoyl peptides inhibited infections by the A/Puerto Rico/8/34 (H1N1) and A/Aichi/2/68 (H3N2) strains of influenza virus. Such HA-inhibitors are promising candidates for novel antiviral drugs. 1765 EPYGFIAFSRAAHSP(4)[0.563 ± 0.310] ARLSPTMVHPNGAQP(2)[1.268 ± 0.179] TFFQVPPRILVGSAS(1)[0.170 ± 0.201] ARLGSVLISSGPSSD(1)[0.261] GRPPDSVFRSRGWLS(1)[0.891 ± 0.280] AWRNMIRYGVDLPAF(1)[0.323 ± 0.092] 6 Phage display (competitive panning) 5 PMID:20476787 Hemagglutinin, HA 6' ganglioside GM3 Not determined. Not determined. X15 fUSE5 phage display library ELISA,Surface plasmon resonance (SPR) In phage ELISA, the absorbance at 492 nm was determined. Data shown were reproduced from the graph and expressed as means ± the standard deviation (n = 3). Besides, clones f1 (displaying ARLSPTMVHPNGAQP), f9 (displaying EPYGFIAFSRAAHSP) and f12 (displaying GRPPDSVFRSRGWLS) were found to have a high affinity by SPR analysis, with dissociation constant (Kd) values of 7.8-30nM against H1 and H3. A docking simulation suggested that, similar to sialic acid, the peptides are recognized by the receptor-binding site in HA, which indicates that these peptides mimic the sialic acid structure. N-stearoyl peptides inhibited infections by the A/Puerto Rico/8/34 (H1N1) and A/Aichi/2/68 (H3N2) strains of influenza virus. Such HA-inhibitors are promising candidates for novel antiviral drugs. 1766 ARMRYVDLPVVSGIS(3)[0.117 ± 0.018] GRVPVFGLSPLFKVE(2)[1.132 ± 0.131] ARLSPTMVHPNGAQP(2)[1.276 ± 0.075] EPYGFIAFSRAAHSP(1)[0.716 ± 0.389] IDIAFSSLALADISR(1)[1.018 ± 0.210] 5 Phage display (competitive panning) 4 PMID:20476787 Hemagglutinin, HA Sialyl Lewis(X), SLX Not determined. Not determined. X15 fUSE5 phage display library ELISA,Surface plasmon resonance (SPR) In phage ELISA, the absorbance at 492 nm was determined. Data shown were reproduced from the graph and expressed as means ± the standard deviation (n = 3). Besides, clones f1 (displaying ARLSPTMVHPNGAQP), f9 (displaying EPYGFIAFSRAAHSP) and f12 (displaying GRPPDSVFRSRGWLS) were found to have a high affinity by SPR analysis, with dissociation constant (Kd) values of 7.8-30nM against H1 and H3. A docking simulation suggested that, similar to sialic acid, the peptides are recognized by the receptor-binding site in HA, which indicates that these peptides mimic the sialic acid structure. N-stearoyl peptides inhibited infections by the A/Puerto Rico/8/34 (H1N1) and A/Aichi/2/68 (H3N2) strains of influenza virus. Such HA-inhibitors are promising candidates for novel antiviral drugs. 1767 ARLSPTMVHPNGAQP(9)[1.276 ± 0.075] IDIAFSSLALADISR(1)[1.018 ± 0.210] 2 Phage display (competitive panning) 5 PMID:20476787 Hemagglutinin, HA 6' ganglioside GM3 Not determined. Not determined. X15 fUSE5 phage display library ELISA,Surface plasmon resonance (SPR) In phage ELISA, the absorbance at 492 nm was determined. Data shown were reproduced from the graph and expressed as means ± the standard deviation (n = 3). Besides, clones f1 (displaying ARLSPTMVHPNGAQP) and f16 (displaying IDIAFSSLALADISR) were found to have a high affinity by SPR analysis, with dissociation constant (Kd) values of 7.8-19nM against H1 and H3. A docking simulation suggested that, similar to sialic acid, the peptides are recognized by the receptor-binding site in HA, which indicates that these peptides mimic the sialic acid structure. N-stearoyl peptides inhibited infections by the A/Puerto Rico/8/34 (H1N1) and A/Aichi/2/68 (H3N2) strains of influenza virus. Such HA-inhibitors are promising candidates for novel antiviral drugs. 1768 QLMHDYR(1)[6.7] LSQSLTR(1)[5.7] RACSKDA(1)[5.1] ANTLRSP(1)[4.8] KHVPKMH(1)[4.6] AHPALPL(1)[3.9] YLTMPTP(1)[2.7] VYLTGPS(1)[2.7] 8 Phage display (common panning) 4 PMID:18500833 α form of poly(L-lactide) crystalline films Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA,Surface plasmon resonance (SPR) The binding of all phage clones, which were identified after four rounds of biopanning against the α form of PLLA films, at a 200 pM phage concentration was analyzed by ELISA. The apparent binding constants (Kapp, 1.0e9 1/M) for films composed of the α form of PLLA were estimated and shown. The value of the library was 1.6e9 1/M. Besides, the real-time analysis of the binding of synthetic peptide (QLMHDYR) freed from the phage particles against the polymer films was performed using a BIAcore X. The binding constant (Ka) for the α form of PLLA was determined to be 6.1e4 1/M. The binding of all phage clones, which were identified after four rounds of biopanning against the R form of PLLA films, at a 200 pM phage concentration was analyzed by ELISA. Therefore, eight prospective clones that showed relatively large binding amounts for the ??? 1769 DYFSSPYYEQLF WPGWHHVPPAVS GHWHHITKVSKQ 3 Phage display (subtractive panning) 5-6 PMID:15379530 Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) To minimize background binding, the phage library was pre-incubated with M-280 streptavidin-coated magnetic beads. Unbound phages were incubated with the SWNHs-magnetic bead complexes. SWNHs-magnetic bead complexes were formed by mixing the sonicated biotinylated SWNHs with streptavidin-coated magnetic beads. Authors selected 33 phage clones from panning cycle 5 and 15 phage clones from panning cycle 6 (total of 48 phage clones), and the pIII tail sequences were identified via standard DNA sequencing techniques. Among the 48 clones, a total of 28 clones (i.e., 15 out of the 33 clones from panning cycle 5 and 13 out of the 15 clones from panning cycle 6) have been shown to display an identical peptide sequence, DYFSSPYYEQLF, that is enriched in polar and aromatic residues. 1770 HSSYWYAFNNKT HTSYWYAFNTKT YTTHVKPFAPSS HAWVDWIRPIH 4 Phage display (common panning) 4 PMID:16402784 Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1771 GETRAPL(7)[0.948 ± 0.058] FPGRPSP(6)[0.902 ± 0.100] YLTMPTP(2)[0.901 ± 0.026] FSWEAFA(1)[0.976 ± 0.029] HLESTPG(1)[0.884 ± 0.034] RHEPPLA(1)[0.857 ± 0.052] GETQCAA(1)[0.836 ± 0.086] HTAQSTA(1)[0.961 ± 0.049] 8 Phage display (common panning) 4 PMID:17492699 Syndiotactic polystyrene film Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA In phage ELISA, the absorbance at 405 nm was determined. Data were reproduced from the graph and shown as means ± standard deviations. The absorbance for negative control was 0.592 ± 0.007. 1772 LAYWEFVFHPQGDDL 1 Phage display (common panning) 3 PMID:7649995 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. X15 phage display library Following three rounds of enrichment and amplification, the 15-mer library was enriched for peptides containing the HPQ epitope in a sequence such as LAYWEFVFHPQGDDL. 1773 PWAWLT PWAWLI 2 Phage display (common panning) 3 PMID:7649995 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. X6 fUSE5 phage dislpay library 1774 LFYYLH(4) VFYYLH(3) RFYYLH(1) AFYYLH(1) HFYYLH(1) PPYYLH(4) PVYYLH(3) GVFYLH(2) RVWYLH(2) GPWYLH(1) QLYFLH(4) QLFFLH(2) QLYRLH(2) [YFW]-[YF]-L-H 13 Phage display (common panning) 2 PMID:7685302 Anti-biotin polyclonal antibody Biotin Not determined. Not determined. f3-6mer phage display library (X6) Each of the phage isolated in the anti-biotin antibody panning were coated on ELISA wells and their ability to bind streptavidin coupled to horseradish peroxidase evaluated. None bound above the background levels of a control phage. 1775 GDWVFI(6) GDWAFI(1) GDFRFT(1) GNFVFI(1) VDWAFI(1) PWPWLG(4) PWLWLQ(1) PWDWLY(1) AFWAWL(2) TWWGYL(1) 9 Phage display (competitive panning) 2 PMID:7685302 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. f3-6mer phage display library (X6) Only phage displaying the GDWVFI and PWPWLG peptides demonstrated biotin-sensitive streptavidin binding in ELISA and micropanning assays. Binding of streptavidin to phage displaying GDWVFI was also confirmed by a BIACORE analysis; both biotin and synthetic HPQ peptide [YGGHPQGG] blocked this binding. Phage displaying GDWVFI could be eluted from streptavidin with either acid or biotin, but not phosphate-buffered saline, whereas phage displaying PWPWLG eluted with acid, biotin and phosphate-buffered saline. The possibility that PWPWLG sticks nonspecifically to plastic was considered; however, it was found that PWPWLG did not bind well unless wells were coated with streptavidin. 1776 SSLRGF(41)[1.087 ± 0.059] 1 Phage display (competitive panning) PMID:8623148 GSI-B4 isolectin Alpha D-Galactose 1HQL, Not determined. f3-6mer phage display library (X6) ELISA In phage ELISA, the absorbance at 505 nm was determined. Data were reproduced from the graph and shown as means ± standard deviations. Phage displaying peptide WAGSTR originated from the random library that had not been biopanned with the absorbance of 0.268 ± 0.028. A phage (displaying PARENT) from which the peptide library was generated is included as a control with the absorbance of 0.260 ± 0.036. The 100-mm tissue culture plates were coated with streptavidin prior to capture of biotinylated GS-1-B4. Phages bound to GS-1-B4 were eluted with melibiose obtained from Sigma. The plates thus prepared were then biopanned against the random peptide library. After biopanning, approximately 100 randomly picked clones were sequenced through the random peptide insertion site to identify potential motifs. Nearly 40% of the phage clones identified from GS-1-B4 biopanning contained peptide SSLRGF. This peptide blocks the binding of GS-1-B4 to pig aortic endothelial cells. The carbohydrate Gal alpha (1,3)Gal competes with the binding of GS-1-B4 to the peptide, suggesting that they may bind the same site. Using a RBC agglutination assay, we show that this peptide inhibits the agglutination of pig RBCs by heat-inactivated human serum at concentrations similar to that of Gal alpha (1,3)Gal. 1777 RNEPPL(3) LWYATE(3) SQMPPL(3) RGAIEP(2) FSAATE(2) ATESAM(1) ATEVVG(1) ATEARE(1) RLDPPI(1) RWDPPI(1) 10 Phage display (common panning) PMID:8623148 Anti-pig Xenoreactive Natural Antibodies, XNAs Gal alpha (1,3)Gal Not determined. Not determined. f3-6mer phage display library (X6) The 100-mm tissue culture plates were coated with streptavidin prior to capture of biotinylated XNAs. The plates thus prepared were then biopanned against the random peptide library. 1778 FGRLVSSIRY(1) TWKTSRISIF(6) 2 Phage display (subtractive panning, competitive panning) 5 PMID:9894906 Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) Immunoglobulin G-binding protein A 1FC2, Not determined. X10 fUSE5 phage display library The decapeptide library was preselected against streptavidin-coated paramagnetic beads to reduce the number of phage clones interacting with streptavidin or with the plastic. The reduced library was then subjected to five selections against IgG Fc. In the first selection, interacting phage peptides were first eluted with protein A and then by lowering the pH. In subsequent selections, phages were elute by lowering the pH. Individual peptide phage clones were found to interact more strongly with IgG Fc than did either the original library or the wild-type phage. It was found that increasing concentrations of protein A competitively reduced the interaction of FARLVSSIRY-phage clone with IgG Fc to the same level as the primary library, indicating that this peptide really interacts with the protein A binding site on IgG Fc. On the other hand, it was only possible to reduce marginally the interaction of TWKTSRISIF-phage clone with IgG Fc using protein A as a competitor. When immunoglobulins from chicken, donkey, human, mouse, swine, rabbit, and sheep were included, peptide phage clones FGRLVSSIRY and TWKTSRISIF interacted strongly with human IgG Fc and porcine IgG and weakly with the immunoglobulins obtained from the other species. 1779 RLWLHRHKLV(1) FARLVSSIRY(1) FGRLVSSIRY(2) TWKTSRISIF(2) 4 Phage display (subtractive panning, competitive panning) 5 PMID:9894906 Fc domain of IgG (IGHG1,IGHG2,IGHG3,IGHG4) Immunoglobulin G-binding protein A 1FC2, Not determined. X10 fUSE5 phage display library The decapeptide library was preselected against streptavidin-coated paramagnetic beads to reduce the number of phage clones interacting with streptavidin or with the plastic. The reduced library was then subjected to five selections against IgG Fc. In the first selection, interacting phage peptides were first eluted with protein A and then by lowering the pH. In subsequent selections, phages were elute by protein A. It was found that increasing concentrations of protein A competitively reduced the interaction of FARLVSSIRY-phage clone with IgG Fc to the same level as the primary library, indicating that this peptide really interacts with the protein A binding site on IgG Fc. On the other hand, it was only possible to reduce marginally the interaction of TWKTSRISIF-phage clone with IgG Fc using protein A as a competitor. When immunoglobulins from chicken, donkey, human, mouse, swine, rabbit, and sheep were included, peptide phage clones FGRLVSSIRY and TWKTSRISIF interacted strongly with human IgG Fc and porcine IgG and weakly with the immunoglobulins obtained from the other species. 1780 EPMTPHQWITLYRSY(0.37) DTPYPWGWLLDEGYD(0.22) FCPPILPYSAWCPVP(0.10) MPVSRLCIELDWCPP(0.10) RGTQEWTELWVSFRA(0.05) RTGHSHDPRSMPKSC(0.05) YGTAPEWVLAFRLAF(0.02) RHRSGTASASKMSPI(0.02) VSPMEIWTTCTWWSS(0.02) SRGSHEWAVLFRFYY(0.02) 10 Phage display (subtractive panning, competitive panning) 4 PMID:10600340 IgG from the cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients Not determined. Not determined. Not determined. f88-15mer phage display library (X15) The cerebrospinal fluids (100 ml, about 5a??a Thirty-six clones were randomly picked and sequenced. The two peptides (DTPYPWGWLLDEGYD and RTGHSHDPRSMPKSC) were not specifically recognized by either cerebrospinal fluids from multiple sclerosis patient or from viral myelitis patient. In contrast, FCPPILPYSAWCPVP, EPMTPHQWITLYRSY, SRGSHEWAVLFRFYY, MPVSRLCIELDWCPP and RGTQEWTELWVSFRA were significantly recognized by cerebrospinal fluids from multiple sclerosis patient but not by cerebrospinal fluids from other neurological diseases. The combination of 4 selected mimotopes (FCPPILPYSAWCPVP, SRGSHEWAVLFRFYY, RGTQEWTELWVSFRA, EPMTPHQWITLYRSY), allowed the detection of specific antibodies in 21 of 60 multiple sclerosis cerebrospinal fluids whereas only 2 of 27 cerebrospinal fluids from patients with other neurological diseases equally recognized the 4 mimotopes. 1781 EPMTPHQWITLYRSY(0.66) SRGSHEWAVLFRFYY(0.09) FCPPILPYSAWCPVP(0.06) RGTQEWTELWVSFRA(0.06) QSPLEDRILRFLSPP(0.06) 5 Phage display (common panning) 3 PMID:10600340 IgG from the cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients Not determined. Not determined. Not determined. f88-15mer phage display library (X15) The cerebrospinal fluids from only one multiple sclerosis patient was used as the target, which was imobilized on to the streptavidin-coated polystyrene petri dishes through biotinylated mouse anti-human IgG monoclonal antibody P5F2F7. The three rounds of biopanning were performed using decreasing amounts of the same cerebrospinal fluids from the multiple sclerosis patient (10, 1, and 0.1 mg of IgG, respectively). Thirty-six clones were randomly picked and sequenced. FCPPILPYSAWCPVP, EPMTPHQWITLYRSY, SRGSHEWAVLFRFYY and RGTQEWTELWVSFRA were significantly recognized by cerebrospinal fluids from multiple sclerosis patient but not by cerebrospinal fluids from other neurological diseases. The combination of 4 selected mimotopes (FCPPILPYSAWCPVP, SRGSHEWAVLFRFYY, RGTQEWTELWVSFRA, EPMTPHQWITLYRSY), allowed the detection of specific antibodies in 21 of 60 multiple sclerosis cerebrospinal fluids whereas only 2 of 27 cerebrospinal fluids from patients with other neurological diseases equally recognized the 4 mimotopes. 1782 EPMTPHQWITLYRSY(0.30) FCPPILPYSAWCPVP(0.15) MPVSRLCIELDWCPP(0.11) QSPLEDRILRFLSPP(0.07) SSRQGLIDCLWSCTH(0.07) SRGSHEWAVLFRFYY(0.04) VSPMEIWTTCTWWSS(0.04) 7 Phage display (common panning) 3 PMID:10600340 IgG from the cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients Not determined. Not determined. Not determined. f88-15mer phage display library (X15) The cerebrospinal fluids from only one multiple sclerosis patient was used as the target, which was imobilized on to the streptavidin-coated polystyrene petri dishes through biotinylated mouse anti-human IgG monoclonal antibody P5F2F7. The three rounds of biopanning were performed using decreasing amounts of the same cerebrospinal fluids from the multiple sclerosis patient (10, 1, and 0.1 mg of IgG, respectively). Thirty-six clones were randomly picked and sequenced. FCPPILPYSAWCPVP, EPMTPHQWITLYRSY, SRGSHEWAVLFRFYY and MPVSRLCIELDWCPP were significantly recognized by cerebrospinal fluids from multiple sclerosis patient but not by cerebrospinal fluids from other neurological diseases. The combination of 4 selected mimotopes (FCPPILPYSAWCPVP, SRGSHEWAVLFRFYY, RGTQEWTELWVSFRA, EPMTPHQWITLYRSY), allowed the detection of specific antibodies in 21 of 60 multiple sclerosis cerebrospinal fluids whereas only 2 of 27 cerebrospinal fluids from patients with other neurological diseases equally recognized the 4 mimotopes. 1783 FCPPILPYSAWCPVP(0.26) MPVSRLCIELDWCPP(0.23) EPMTPHQWITLYRSY(0.20) QSPLEDRILRFLSPP(0.06) SRGSHEWAVLFRFYY(0.03) VSPMEIWTTCTWWSS(0.03) SSRQGLIDCLWSCTH(0.03) 7 Phage display (common panning) 3 PMID:10600340 IgG from the cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients Not determined. Not determined. Not determined. f88-15mer phage display library (X15) The cerebrospinal fluids from only one multiple sclerosis patient was used as the target, which was imobilized on to the streptavidin-coated polystyrene petri dishes through biotinylated mouse anti-human IgG monoclonal antibody P5F2F7. The three rounds of biopanning were performed using decreasing amounts of the same cerebrospinal fluids from the multiple sclerosis patient (10, 1, and 0.1 mg of IgG, respectively). Thirty-six clones were randomly picked and sequenced. FCPPILPYSAWCPVP, EPMTPHQWITLYRSY, SRGSHEWAVLFRFYY and MPVSRLCIELDWCPP were significantly recognized by cerebrospinal fluids from multiple sclerosis patient but not by cerebrospinal fluids from other neurological diseases. The combination of 4 selected mimotopes (FCPPILPYSAWCPVP, SRGSHEWAVLFRFYY, RGTQEWTELWVSFRA, EPMTPHQWITLYRSY), allowed the detection of specific antibodies in 21 of 60 multiple sclerosis cerebrospinal fluids whereas only 2 of 27 cerebrospinal fluids from patients with other neurological diseases equally recognized the 4 mimotopes. 1784 NACYVDLFLGASVCP(0.46)[0.07] SSAKSHCYAFCSGLP(0.31)[0.25] HCRKVTGSDYLLCGL(0.05)[0.05] 3 Phage display (subtractive panning, competitive panning) 4 PMID:10600340 IgG from the cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients Not determined. Not determined. Not determined. f88-15mer phage display library (X15) ELISA IgG immunoreactivity of MS sera against the selected phage-displayed peptides was detected by ELISA on immobilized phages with a pool of five MS sera diluted to 1:100. The results are expressed as the difference between the OD obtained at 492 nm with MS sera and that with non-MS sera. The serum (20 ml, about 200 mg of IgG) from multiple sclerosis patients was used as the target, which was imobilized on to the streptavidin-coated polystyrene petri dishes through biotinylated mouse anti-human IgG monoclonal antibody P5F2F7. For each round of panning, serum from a different multiple sclerosis patient was used. In the fourth round, the amplified eluate was incubated with 100 ml of a pool of sera from five non-multiple sclerosis patients before reacting with dish-bound serum IgG from multiple sclerosis patient. Thirty-six clones were randomly picked and sequenced. Compared to control peptide, NACYVDLFLGASVCP and HCRKVTGSDYLLCGL were not specifically recognized by individual multiple sclerosis sera whereas SSAKSHCYAFCSGLP was significantly recognized by only 3 of 37 sera. 1785 NACYVDLFLGASVCP(0.50)[0.07] HCRKVTGSDYLLCGL(0.33)[0.05] SSAKSHCYAFCSGLP(0.06)[0.25] SPLNLCFERCFTSPN(0.06)[ND] SHPTLMPPLARMPSL(0.03)[ND] 5 Phage display (subtractive panning, competitive panning) 4 PMID:10600340 IgG from the cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients Not determined. Not determined. Not determined. f88-15mer phage display library (X15) ELISA IgG immunoreactivity of MS sera against the selected phage-displayed peptides was detected by ELISA on immobilized phages with a pool of five MS sera diluted to 1:100. The results are expressed as the difference between the OD obtained at 492 nm with MS sera and that with non-MS sera. The serum (20 ml, about 200 mg of IgG) from multiple sclerosis patients was used as the target, which was imobilized on to the streptavidin-coated polystyrene petri dishes through biotinylated mouse anti-human IgG monoclonal antibody P5F2F7. For each round of panning, serum from a different multiple sclerosis patient was used. In the fourth round, the amplified eluate was incubated with 100 ml of a pool of sera from five non-multiple sclerosis patients before reacting with dish-bound serum IgG from multiple sclerosis patient. Thirty-six clones were randomly picked and sequenced. Compared to control peptide, NACYVDLFLGASVCP and HCRKVTGSDYLLCGL were not specifically recognized by individual multiple sclerosis sera whereas SSAKSHCYAFCSGLP was significantly recognized by only 3 of 37 sera. 1786 AGPDCSSLPNSMRCS(6) GSRPSPIGSKWALQP(2) TSPHRPSAIGVLPPL(2) EKERRPSPIGTATLL(1) GKRPSCTGCSYELAQ(1) GGRHPSPMGTFDTQT(1) STKPSSIGTLHTSPS(1) AVSGIGTQTSIPYPS(1) LTTEPDTTKFWPAPS(1) SPPLQCSQYPNSLRC(1) QQPVSSWVFVSSDAI(1) SRPSP 11 Phage display (subtractive panning) 5 PMID:10848928 Anti-IgE polyclonal antibody IgG Ig epsilon chain C region Not determined. Not determined. f88-15mer phage display library (X15) Ten μg of biotinylated goat antihuman IgG (capture antibody) incubated with 1 mg of streptavidin-coated Dynabeads M-280. The beads were then washed five times in Tris-buffered saline (TBS) pH 7.4 and resuspended in 100mL TBS. This procedure was repeated with a second aliquot (1 mg) of beads. Ten mg of human IgG anti-IgE (selector antibody) was incubated with the first aliquot of beads for 2 hours at room temperature, and the beads were then washed four times in TBS. From the amplified phage-peptide library 60μL was added to the control beads and the mixture was incubated for 3 h at room temperature. This was done to remove any phage-peptides binding to the biotinylated antihuman IgG. The phage-peptide library, treated in this way, was then incubated with the IgG anti-IgE coated beads for 3 h at room temperature. 1787 VAWCTIFLCLDV[0.22] ADFCEGKDMIDWVYCRLY[2.5] FWFCDRIAWYPQHLCEFL[1.9] FRNCEPWMLRFGCNPR[4.8] 4 Phage display (common panning) 2 PMID:11934284 Human serum albumin Not determined. Not determined. Not determined. TN phage display library pool Fluorescence anisotropy Fluorescence anisotropy measurements were performed in various conditions. It was discovered that many of the peptides bound better at a lower pH (3 mM phosphate, pH 6.2) and in the absence of added salt. The dissociation constants (KD, μM) of this condition were shown. The pooled phage libraries of TN6-6, TN10-9, and TN12-1 were used in selections against human serum albumin (HSA) immobilized directly on polystyrene microtiter plates. After two rounds of selection, phage peptide selection on directly immobilized HSA yielded several phage isolates (232, 234, 236, and 238) that showed positive HSA binding by ELISA. Phage isolate 232 (VAWCTIFLCLDV) showed the highest ELISA signal. Although in solution DX-232 bound HSA with the highest affinity, the DX-232-Sepharose column performance was bad, binding no detectable HSA. In contrast, the DX-236(FWFCDRIAWYPQHLCEFL)-Sepharose column was the best performer and quantitatively bound the entire 1 mg injection (0.15 μmol). At higher HSA loads in the no salt, pH 6.2 buffer, the DX-236-Sepharose column (0.35 mL) bound upward of 4 mg of HSA, which corresponds to a dynamic capacity of greater than 11 mg/mL. The DX-236 affinity column bound HSA from human serum with a greater specificity than does Cibacron Blue agarose beads. DX-236 also bound well to several mammalian serum albumins. 1788 FKICDQWFCLMP[1.8] HVGCNNALCMQY[17] WKVCDHFFCLSP[18] NHGCWHFSCIWD[1.9] 4 Phage display (subtractive panning, competitive panning) 4 PMID:11934284 Human serum albumin Not determined. Not determined. Not determined. TN6-6 phage display library Fluorescence anisotropy Fluorescence anisotropy measurements were performed in various conditions. It was discovered that many of the peptides bound better at a lower pH (3 mM phosphate, pH 6.2) and in the absence of added salt. The dissociation constants (KD, μM) of this condition were shown. The TN6-6 phage library was processed to remove bead binders and then screened against caprylate-biotinylated-HSA in solution for 1h; later on, HSA-binding phage were captured by the addition of magnetic streptavidin beads for 15 min. Phage isolates 298 (NHGCWHFSCIWD) showed the highest ELISA signal. 1789 DWDCVTRWANRDQQCWGP[9.5] DWDCVTRWANRDQQCWAL[13] DWDCVTDWANRHQHCWAL[6.7] DWQCVKDWANRRRGCMAD[17] RNMCKFSWIRSPAFCARA[0.9] 5 Phage display (subtractive panning, competitive panning) 4 PMID:11934284 Human serum albumin Not determined. Not determined. Not determined. TN12-1 phage display library Fluorescence anisotropy Fluorescence anisotropy measurements were performed in various conditions. It was discovered that many of the peptides bound better at a lower pH (3 mM phosphate, pH 6.2) and in the absence of added salt. The dissociation constants (KD, μM) of this condition were shown. The TN12-1 phage library was processed to remove bead binders and then screened against caprylate-biotinylated-HSA in solution for 1h; later on, HSA-binding phage were captured by the addition of magnetic streptavidin beads for 15 min. Phage isolates 321 (RNMCKFSWIRSPAFCARA) showed the highest ELISA signal. DX-321 had a KD value less than 2μM in no salt buffer at pH 6.2. It also showed a strong preference for albumin from humans. 1790 NCVSPYWCEPLAPSARA 1 Phage display (competitive panning) PMID:12890473 GSI-B4 isolectin Alpha D-Galactose 1HQL, Not determined. XCX15 phage display library Phages bound to GS-1-B4 were eluted with melibiose obtained from Sigma. After biopanning, 18 clones were randomly picked. On four out of them was performed the ELISA to compare the binding activity. All clones selected were able to bind the lectin BS-I-B4. A phage bearing the peptide NCVSPYWCEPLAPSARA has been identified to bind the lectin strongly. Melibiose was able to inhibit the binding of the human natural anti-alpha-Gal antibody to the peptide competitively. The peptide also inhibited the agglutination of pig RBCs caused by human natural antibody or lectin BS-I-B4. 1791 HLEPLIS(10) 1 Phage display (common panning) 1 PMID:15472891 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. Ph.D.-7 phage display library (X7) In total, 10 clones were sequenced. 1792 HLEPLIS(6) 1 Phage display (common panning) 2 PMID:15472891 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. Ph.D.-7 phage display library (X7) In total, 6 clones were sequenced. 1793 TESMLPS(2) 1 Phage display (competitive panning) 1 PMID:15472891 SB-365579 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) All incubation steps were carried out at room temperature. NeutrAvidin-coated plates were used to capture SB-365579. To reduce nonspecific binding, a blocking step was performed using 5% skim milk. All of the compounds were initially dissolved in dimethyl formamide (DMF) (22 mM drug stock solutions) and then diluted in TBSTM (tris-buffered saline, pH 7.5, with 0.1% Tween-20 and 5% skim milk) to a final concentration of 22 mM. Bound phage from the drug-coated wells were eluted with 200 μl of 220 μM SB-365579 solution in TBSTM. After the fisrt round of panning, 29 clones were sequenced. However, many sequences were not given in the original paper. 1794 HLEPLIS(6) ATLAPFT(2) 2 Phage display (competitive panning) 2 PMID:15472891 SB-365579 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) All incubation steps were carried out at room temperature. NeutrAvidin-coated plates were used to capture SB-365579. To reduce nonspecific binding, a blocking step was performed using 5% skim milk. All of the compounds were initially dissolved in dimethyl formamide (DMF) (22 mM drug stock solutions) and then diluted in TBSTM (tris-buffered saline, pH 7.5, with 0.1% Tween-20 and 5% skim milk) to a final concentration of 22 mM. Bound phage from the drug-coated wells were eluted with 200 μl of 220 μM SB-365579 solution in TBSTM. After the second round of panning, 21 clones were sequenced. However, many sequences were not given in the original paper. 1795 LPLTPLP(8) ATLAPFT(6) HLEPLIS(2) 3 Phage display (competitive panning) 3 PMID:15472891 SB-365579 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) All incubation steps were carried out at room temperature. NeutrAvidin-coated plates were used to capture SB-365579. To reduce nonspecific binding, a blocking step was performed using 5% skim milk. All of the compounds were initially dissolved in dimethyl formamide (DMF) (22 mM drug stock solutions) and then diluted in TBSTM (tris-buffered saline, pH 7.5, with 0.1% Tween-20 and 5% skim milk) to a final concentration of 22 mM. Bound phage from the drug-coated wells were eluted with 200 μl of 220 μM SB-365579 solution in TBSTM. After the third round of panning, 26 clones were sequenced. However, many sequences were not given in the original paper. 1796 TPSPERP(12) LPLTPLP(7) ATLAPFT(6) ATPLWLK(1) 4 Phage display (competitive panning) 1 PMID:15472891 SB-365580 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) All incubation steps were carried out at room temperature. NeutrAvidin-coated plates were used to capture SB-365580. To reduce nonspecific binding, a blocking step was performed using 5% skim milk. All of the compounds were initially dissolved in dimethyl formamide (DMF) (22 mM drug stock solutions) and then diluted in TBSTM (tris-buffered saline, pH 7.5, with 0.1% Tween-20 and 5% skim milk) to a final concentration of 22 mM. Bound phage from the drug-coated wells were eluted with 200 μl of 220 μM SB-365580 solution in TBSTM. After the fisrt round of panning, 27 clones were sequenced. However, there was one sequence not given in the original paper. 1797 TPSPERP(5) LPLTPLP(8) ATLAPFT(3) ATPLWLK(1) 4 Phage display (competitive panning) 2 PMID:15472891 SB-365580 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) All incubation steps were carried out at room temperature. NeutrAvidin-coated plates were used to capture SB-365580. To reduce nonspecific binding, a blocking step was performed using 5% skim milk. All of the compounds were initially dissolved in dimethyl formamide (DMF) (22 mM drug stock solutions) and then diluted in TBSTM (tris-buffered saline, pH 7.5, with 0.1% Tween-20 and 5% skim milk) to a final concentration of 22 mM. Bound phage from the drug-coated wells were eluted with 200 μl of 220 μM SB-365580 solution in TBSTM. After the second round of panning, 17 clones were sequenced. 1798 TPSPERP(9) LPLTPLP(8) ATLAPFT(8) 3 Phage display (competitive panning) 3 PMID:15472891 SB-365580 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) All incubation steps were carried out at room temperature. NeutrAvidin-coated plates were used to capture SB-365580. To reduce nonspecific binding, a blocking step was performed using 5% skim milk. All of the compounds were initially dissolved in dimethyl formamide (DMF) (22 mM drug stock solutions) and then diluted in TBSTM (tris-buffered saline, pH 7.5, with 0.1% Tween-20 and 5% skim milk) to a final concentration of 22 mM. Bound phage from the drug-coated wells were eluted with 200 μl of 220 μM SB-365580 solution in TBSTM. After the third round of panning, 27 clones were sequenced. However, some sequences were not given in the original paper. 1799 KVWVLPI(51) 1 Phage display (competitive panning) 3 PMID:15574836 70 kDa heat shock cognate protein 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding ability of the high-frequency peptides was confirmed by ELISA. Absorbances at 405 nm were measured (data not shown). Normalized data were used to quantify the results. The heptapeptide KVWVLPI had the highest binding ability to the target protein Vr130. The recombinant proteins were designated Vr130 for polypeptides containing the C-terminal 30 kDa domain of VrHsc70-1. The recombinant proteins were purified on a metal-affinity column and then cleaved with thrombin to remove the His tag. The Vr130 recombinant proteins was used to select VrHsc70-binding heptapeptides using phage display. The bound phage was eluted with a 1 ml solution of the free target protein. Among 125 different clones selected with Vr130, the heptapeptide KVWVLPI had the highest frequency(40.8%). The binding ability of the high-frequency peptides was confirmed by ELISA. 1800 KLWVIPQ(54) 1 Phage display (competitive panning) 3 PMID:15574836 70 kDa heat shock cognate protein 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding ability of the high-frequency peptides was confirmed by ELISA. Absorbances at 405 nm were measured (data not shown). Normalized data were used to quantify the results. The peptide KLWVIPQ bound strongly to Vr230, but exhibited <5% binding ability towards Vr130. Conversely, KVWVLPI exhibited 13% binding ability towards Vr230. The recombinant proteins were designated Vr230 for polypeptides containing the C-terminal 30 kDa domain of VrHsc70-2. The recombinant proteins were purified on a metal-affinity column and then cleaved with thrombin to remove the His tag. The Vr230 recombinant proteins was used to select VrHsc70-binding heptapeptides using phage display. The bound phage was eluted with a 1 ml solution of the free target protein. Among 123 different clones of Vr230, the heptapeptide KLWVIPQ had the highest frequency (43.9%). The binding ability of the high-frequency peptides was confirmed by ELISA. 1801 YAPLSRL(48) KLWVIPQ(33) 2 Phage display (competitive panning) 3 PMID:15574836 70 kDa heat shock cognate protein 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding ability of the high-frequency peptides was confirmed by ELISA. Absorbances at 405 nm were measured (data not shown). The heptapeptide KLWVIPQ displayed about 60% of the binding ability to Vr330 as compared with YAPLSRL, which had the highest binding ability. The relative binding ability of Vr330 to any other low-frequency heptapeptide was also <5%. The recombinant proteins were designated Vr330 for polypeptides containing the C-terminal 30 kDa domain of VrHsc70-3. The recombinant proteins were purified on a metal-affinity column and then cleaved with thrombin to remove the His tag. The Vr330 recombinant proteins was used to select VrHsc70-binding heptapeptides using phage display. The bound phage was eluted with a 1 ml solution of the free target protein. Among the 116 different clones of Vr330, 41.4% were peptides with a sequence of YAPLSRL and 28.4% had the peptide sequence of KLWVIPQ. The heptapeptide KLWVIPQ displayed about 60% of the binding ability to Vr330 as compared with YAPLSRL, which had the highest binding ability confirmed by ELISA. 1802 EPGHDAVP E-X(2)-H 1 Phage display (competitive panning) 4 PMID:16398491 Cobalt ion, Co(2+) Not determined. Not determined. Not determined. Type 8 phage display library (X8) The selection of type 8 phage with affinity toward Co2+ was performed by incubating the type 8 phage library (~e10 pfu) with Co2+ immobilized on 200 μL of Chelating Sepharose Fast Flow gel in TBS buffer (pH 7.5) with 0.15% tween-20. The bound phage were washed 10 times with the incubation buffer and then eluted with 50 mM histidine. 1803 WHWRLPS(10)[2.290 ± 0.094] HKPLQIW(5)[0.363 ± 0.088] QAHTVGK(4)[0.834 ± 0.451] YVHRLPP(2)[0.555 ± 0.078] HYTRHSA(1)[0.373 ± 0.114] 5 Phage display (common panning) 3 PMID:16413648 Microcystin-LR, MC-LR Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Affinities for MC-LR of the five different selected clones were measured by ELISA. Absorbances at 410 nm were reproduced from the bar graph and expressed as the means of three determinations ± sample standard deviations. L7 represented original linear 7 peptides library and its absorbance was 0.256 ± 0.266. Clone L7-3 (displaying WHWRLPS) expressed the highest affinity for MC-LR as compared to the original library and their negative controls. The affinity of selected phage to MC-LR were tested through an immunoprecipitation assay. The results showed that all selected phage combined with MC-LR except the YVHRLPP-bearing phage. 1804 GRIQRWWVMFLL(1) IISRAGVVRGNE(1) ADYLSRWGSIRN(1) ETYVFDNHFHAP(1) TVSEVASTSNLD(1) SRVHRAVLNGVS(1) RPPGVVRRYALG(1) VRSWEEQARVTT(1) RAFIASRRIKRP(1) VSQRVGFITSAP(1) AGAGGGNVPVCS(1) RESTLKGTSRAV(1) AGLRLKKAAIHR(1) SSLLRAVPEPTG(1) EWDITTECTVTF(1) 15 Bacterial display (common panning) 10 PMID:16599553 Petri dish (polystyrene, PS) Not determined. Not determined. Not determined. FliTrx bacterial display library (X12) ELISA The mutant-type GSTs fused with PS-tags no. 16, 17, 19, and 23 (SRVHRAVLNGVS, RPPGVVRRYALG, RAFIASRRIKRP and AGLRLKKAAIHR) were adsorbed to the polystyrene latex beads and the percentage was 80% or higher, whereas those for the wild-type GST and the GST-PS1 were only 50%. RAFIASRRIKRP and AGLRLKKAAIHR showed a strong affinity to PS latex beads with relatively high residual activities. 1805 KGLRGWREMISL(5) LDPGAMRTIVRP(3) QLVEGVRHRIWN(2) 3 Bacterial display (common panning) 5 DOI:10.1016/j.molcatb.2003.12.019 96-well immunoassay plate (polystyrene, PS) Not determined. Not determined. Not determined. FliTrx bacterial display library (X12) The selected peptides were commonly rich in hydrophobic amino acid residues and had two or three basic amino acid residues. Adsorption and desorption experiments for one of the selected peptide (KGLRGWREMISL) showed that it was well and irreversibly adsorbed onto PS latex particles. The most frequent sequence was LGP*AAR*FGDE, where the asterisks represent stop codons. This sequence is not included because of the insertions of stop codons. 1806 LPPWQRQ(9)[0.476 ± 0.014] HPERATL(8)[0.492 ± 0.031] KPRMPPR(4)[0.500 ± 0.028] HDHRYPK(2)[0.494 ± 0.007] HPRWHTP(1)[0.455 ± 0.013] QLKTGLA(1)[0.423 ± 0.033] GKPMPPM(1)[0.337 ± 0.025] THLPWQT(1)[0.382 ± 0.015] HPWWRPS(1)[0.440 ± 0.012] HALGPSS(1)[0.472 ± 0.019] HAIYPRH(1)[0.477 ± 0.004] HKPDANR(1)[0.483 ± 0.018] AITRSPA(1)[0.357 ± 0.014] FPGHSGP(1)[0.400 ± 0.013] NKNYIQH(1)[0.350 ± 0.021] 15 Phage display (common panning) 3 PMID:17154206 Isotactic poly(methyl methacrylate), st-PMMA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA A triplicate ELISA was performed to measure the affinities of all selected clones to conditioned st-PMMA. Absorbances at 405 nm were determined, reproduced from the graph and shown as means ± standard deviations. The absorbance for negative control was 0.120 ± 0.012. st-PMMA films, approximately 40-nm thick, were spincoated on glass slides and immersed in Tris-buffered saline (TBS, Tris=tris(hydroxymethyl)aminomethane; pH 7.4) for 15 h (conditioning). The relative amount of phages bound to the PMMA films is quantified by ELISA. Significantly, for many clones, greater amounts bind to target films than to reference films. The amounts bound to reference films were almost the same within experimental error. 1807 HPERATL(10)[0.492 ± 0.031] LPPWQRQ(3)[0.476 ± 0.014] HPVHPHR(1)[0.507 ± 0.024] 3 Phage display (common panning) 4 PMID:17154206 Isotactic poly(methyl methacrylate), st-PMMA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA A triplicate ELISA was performed to measure the affinities of all selected clones to conditioned st-PMMA. Absorbances at 405 nm were determined, reproduced from the graph and shown as means ± standard deviations. The absorbance for negative control was 0.120 ± 0.012. st-PMMA films, approximately 40-nm thick, were spincoated on glass slides and immersed in Tris-buffered saline (TBS, Tris=tris(hydroxymethyl)aminomethane; pH 7.4) for 15 h (conditioning). The relative amount of phages bound to the PMMA films is quantified by ELISA. Significantly, for many clones, greater amounts bind to target films than to reference films. The amounts bound to reference films were almost the same within experimental error. 1808 HPERATL(14)[0.492 ± 0.031] LPPWQRQ(1)[0.476 ± 0.014] 2 Phage display (common panning) 5 PMID:17154206 Isotactic poly(methyl methacrylate), st-PMMA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA A triplicate ELISA was performed to measure the affinities of all selected clones to conditioned st-PMMA. Absorbances at 405 nm were determined, reproduced from the graph and shown as means ± standard deviations. The absorbances for negative control was 0.120 ± 0.012. st-PMMA films, approximately 40-nm thick, were spincoated on glass slides and immersed in Tris-buffered saline (TBS, Tris=tris(hydroxymethyl)aminomethane; pH 7.4) for 15 h (conditioning). The relative amount of phages bound to the PMMA films is quantified by ELISA. Significantly, for many clones, greater amounts bind to target films than to reference films. The amounts bound to reference films were almost the same within experimental error. 1809 HPERATL(30)[0.492 ± 0.031] LPPWQRQ(10)[0.476 ± 0.014] HPRWHTP(1)[0.455 ± 0.013] HPRLGLA(1)[0.413 ± 0.033] 4 Phage display (common panning) 8 PMID:17154206 Isotactic poly(methyl methacrylate), st-PMMA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA A triplicate ELISA was performed to measure the affinities of all selected clones to conditioned st-PMMA. Absorbances at 405 nm were determined, reproduced from the graph and shown as means ± standard deviations. The absorbances for negative control was 0.120 ± 0.012. st-PMMA films, approximately 40-nm thick, were spincoated on glass slides and immersed in Tris-buffered saline (TBS, Tris=tris(hydroxymethyl)aminomethane; pH 7.4) for 15 h (conditioning). The relative amount of phages bound to the PMMA films is quantified by ELISA. Significantly, for many clones, greater amounts bind to target films than to reference films. The amounts bound to reference films were almost the same within experimental error. 1810 MPLQRLFDGASP(2)[1.308 ± 0.033/1.534 ± 0.047] AYPPSLYDGYNP(1)[1.134 ± 0.027] STHAKLSDGSNP(1)[1.221 ± 0.024] DDRLTSRDGSNP(1)[1.284 ± 0.060] YQLRPNAESLRF(1)[1.161 ± 0.043] GQPLIAHSTKLL(1)[1.534 ± 0.044] YALTPVPSSIRT(1)[1.527 ± 0.043] LFDGSNP 7 Phage display (common panning) 3 PMID:18485511 Anti-E2 protein monoclonal antibody HQ06 E2 protein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The selected peptides for target binding was detected by phage ELISA. Three independent assays were carried out. Absorbances at 490 nm were determined, reproduced from the graph and shown as means of three independent assays ± standard deviations. Phage ELISA results showed that the selected eight phage clones all showed specific reactivity with HQ06 (OD490 nm > 1.10), but not with anti-porcine IFN-γ mAb (OD490 nm < 0.15, data not shown). The present study describes the identification of a linear B-cell epitope at the N-terminus of the E2 protein by screening a phage-displayed random 12-peptide library with the neutralizing monoclonal antibody HQ06 directed against the E2 protein. HQ06 was found to bind to the phages displaying a consensus motif LFDGSNP, which is highly homologous to 772LFDGTNP778 of the CSFV polyprotein, locating on the border between antigenic domains B/C and A of the E2 protein. 1811 APWHLSSQYSRT NLAPIKVSLTSL SNQIPSSARAFI TALATSSTYDPH 4 Phage display (common panning) 5 PMID:18565751 Sulfated Lewis X Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Panning was carried out in a 96-well Streptavidin High Binding Capacity coated microplate or 20 μL of streptavidin agarose bead slurry in a disposable column. 1812 FAAPMRTVQKID RNRRSIQRPMIS 2 Phage display (common panning) 5 PMID:18565751 Sulfated Lewis A Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Panning was carried out in a 96-well Streptavidin High Binding Capacity coated microplate or 20 μL of streptavidin agarose bead slurry in a disposable column. The synthesized peptide FAAPMRTVQKID displayed a consistently high binding affinity and specificity against the cognate HSO3-LeA. The dimeric FAAPMRTVQKID peptide has a low micromolar affinity against the sulfated Lewis A, which is even more specific than an antibody. 1813 WDANGKT(1) ASSLNIA(4) LAPQKLP(1) VSAAPYP(1) AAANVWH(1) AYPGFAL(1) TATITTK(1) 7 Phage display (in vivo) 5 PMID:10204780 Mouse skeletal muscles Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Initially, three rounds of in vitro selection were accomplished on differentiated murine C2C12 myotubes. Phage selected after three in vitro rounds on C2C12 mouse myotubes were injected intravenously into mice to screen for muscle binding phage in vivo. Skeletal muscles were harvested, and the phage rescued from the tissues were amplified and rescreened in a second round. After five rounds of enrichment, phage were plated, 10 random phage clones were picked from the plate. Sequencing the corresponding coding region in the viral DNA from muscle-localizing phage revealed one dominant peptide, ASSLNIA, representing 40% of the clones. Phage carrying this peptide showed a 9- to 20-fold (depending on control tissue) increase in muscle selectivity compared with phage with no insert. When the injected individual phage clone was localized by immunohistochemistry, it was found within focal areas of the membrane of myofibers. Thus, the peptide identified represents a ligand that is capable of accessing skeletal and cardiac muscle from the lumen of blood vessels and could therefore readily be exploited for targeted delivery to muscle cells. 1814 TPAWLDPPT(12) PTAQRWRFR(1) 2 Phage display (common panning) 3 PMID:18502181 Anti-human DAF monoclonal antibody DG3 Complement decay-accelerating factor Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA A total of 100 clones were randomly chosen after the third round of screening and were subjected to ELISA. Thirteen clones were found to react strongly with DG3. The specificity of the two different peptides to immobilized DAF was confirmed by competitive ELISA. The results showed that phage 2 harboring peptide sequences TPAWLDPPT, but not phage 1 harboring peptide sequences PTAQRWRFR , inhibited the binding of DG3 to immobilized DAF in a concentration-dependent manner, demonstrating the specificity of phage 2 to DG3. Virions of the phage library were incubated for 1 h at room temperature with 10 μg of affinitypurified anti-DAF DG3 antibodies. To capture the phage-antibody complexes, 0.5μg/mL biotinylated goat anti-mouse IgG was incubated for 30 min with the antibody-phage complexes. The Petri dish coated with 3μg streptavidin/mL was washed four times and the phage-antibody complexes were added and incubated for 90 min at 37 °C. A total of 100 clones were randomly chosen after the third round of screening and were subjected to ELISA. Thirteen clones were found to react strongly with DG3. All these phage clones were sequenced. These 13 clones comprised only two different sequences. One sample was an independent clone (PTAQRWRFR, frequency 1/13, phage 1), and the other 12 shared the same sequence (TPAWLDPPT, frequency 12/13, phage 2). Two isolated sequences did not show obvious homology. The amino acid sequence similarity between the selected clones and DAF was investigated. Although no absolute consensus sequences were observed between phage 2 and the SCR3 of DAF, phage 2 showed sequence similarity with amino acids 210-218 of DAF SCR3, To determine whether the peptide TPAWLDPPT mimics the structure of the DG3-binding epitope of DAF, competitive binding of DG3 and the peptide TPAWLDPPTwith immobilized DAF was investigated by ELISA. The binding of DG3 to DAF was inhibited by adding the peptide TPAWLDPPT in a concentration-dependent manner, while the control peptide (DAF SCR3 homologous peptide, SVQWSDPPT) had no effect on the binding of DG3 to DAF. These data support that the peptide TPAWLDPPT mimics the DG3-binding epitope of DAF, and that DG3 recognizes a nonlinear constrained epitope. 1815 RSNTRMTARQHRSANHKSTQRARS(8) RSVFLPSILGWRSRLDDQGVAARS(2) RSTRNKHTTARRSVAPGIGEPSRS(3) RSIMHVRLRARRSARHMKDADPRS(1) RSPIIIRSRINRSHGRTKATPARS(1) RSRGLRNILMLRSYDSRSMRPHRS(2) RSTRRGTHNKDRS(4) RSTVPKRHPKDRS(1) RSIAKKTHNKQRS(1) RSYDSRSMRPHRS(1) RSTASRHTEPHRS(1) 11 Bacterial display (common panning) PMID:10618196 Zinc oxide, ZnO Not determined. Not determined. Not determined. X9 E. coli bacterial display library ZnO and bacteria expressing the random peptide library in FimH were mixed and allowed to adhere to each other at room temperature with gentle agitation. Centrifugation was then performed, and the ZnO and any adhering bacteria were recovered and inoculated into Luria-Bertani medium containing the appropriate antibiotics. After overnight incubation, exponentially growing cultures were established, and the enrichment procedure was repeated. The sequences selected exhibited various degrees of affinity and specificity towards ZnO. Competitive binding experiments revealed that the sequences recognized only the oxide form of Zn. Interestingly, one of the inserts, RSNTRMTARQHRSANHKSTQRARS, exhibited significant homology to a specific sequence in a putative zinc-containing helicase, which suggests that searches such as this one may aid in identifying binding motifs in nature. 1816 CNRPFIPTC 1 Phage display (subtractive panning, competitive panning) 4 PMID:12565725 Prothrombin Hirudin variant-2 1A2C,1A46,1A5G,1A61,1B5G,1K21,1K22,1NRS,1VZQ,1W7G,1WAY,2C8W,2C8X,2C8Y,2C8Z,2C90,2C93,2R2M,2ZFQ,2ZFR,2ZG0,2ZHE,2ZHF,2ZHW,3BIU,3BIV,3F68,3HTC,4HTC,1A4W,1DOJ,1TMB,1TMU,3HAT,3P17,3QTO,3QTV,3QWC,3QX5,3RLW,3RLY,3RM0,3RM2,3RML,3RMM,3RMN,3RMO,3SHA,3SHC,3SI3,3SI4,3SV2,3T5F,3TU7,3U98,3U9A,3UWJ,4BAO,4E7R,4LOY,4N3L,4NZE, Not determined. Ph.D.-C7C phage display library (CX7C) Phages were added to half of the blocked streptavidin magnetic particles. The phage supernatant was then transferred to the thrombin-bound magnetic particles coated with streptavidin and incubated overnight at room temperature. In the first round, phages bound to thrombin were eluted with 50 μl of 0.1 M glycine, pH 2, for 15 min at room temperature and immediately neutralised with 125 μl of 1 M Tris-HCl, pH 8. The titre of the eluted phages was estimated and half of the eluted fraction was used for amplification. Three additional rounds of panning were performed. During this last three rounds, the thrombin-bound phages were eluted with 8 mg r-hirudin (Hoechst-Marion-Rousell, AGS Frankfurt, Germany). This peptide, thrombin-inhibiting peptide (TIP), is a full competitive inhibitor of thrombin with an inhibition constant (Ki) of 0.4974 mM. It lengthened the thrombin time and inhibited thrombin-induced platelet activation and the platelet release reaction, both in a dose-dependent manner. It also reduced platelet adhesion onto a human microvascular endothelial matrix in the parallel plate flow chamber under both arterial and venous shear conditions. 1817 TSFAEYWNLLSP(6)[3.4][3.3] LTFEHYWAQLTS(6)[NT][20] DDWFQRVWSPLM(1)[NT][NT] RYEFLDYWSQLH(1)[NT][NT] VPRSAPTLWLGT(1)[NT][NT] F-X(3)-W-X(2)-L 5 Phage display (common panning) 4 PMID:19255450 p53-binding domains of MDM2 Cellular tumor antigen p53 1YCR,4HFZ, 3EQS,3LNZ,3G03,3JZR,3JZS, Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR),Isothermal titration calorimetry (ITC) A surface plasmon resonance (SPR)-based competition assay was performed with (15–29)p53 as the competitor. The Kd value (nM) of synthesized peptides was shown. Besides, the direct interaction between p53-binding domains of MDM2 (referred to thereafter as (syn)MDM2) and the peptide (TSFAEYWNLLSP) were quantified at 25°C, using a MicroCal VP-ITC microcalorimeter. The binding affinity (Kd, nM) was shown. NT denotes not tested. The p53-binding domains of MDM2 (referred to thereafter as (syn)MDM2) and their site-specifically biotinylated forms were chemically synthesized using native chemical ligation. In fact, biotin-(syn)MDM2 was used as bait against the phage display library. Several MDM2 structures have been determined in complexes with PMI (PDB ID code 3EQS and 3LNZ) and pDI (PDB ID code 3G03, 3JZR and 3JZS). The basic procedures for library screening are as follows: (i) incubate input phage (10 μL) with 400 μL of 10 nM biotin (syn)MDM2 for 60 min before adding phage-target solution to 50 μL of streptavidin-agarose resin (Pierce) for affinity capturing; (ii) wash unbound phage; (iii) elute bound phage with 1 mM (15–29) p53; (iv) amplify the eluate and collect phage for the next round of panning; (v) repeat steps 1– 4; (vi) sequence selected binding clones according to the procedures recommended by the manufacturer. Two consensus sequences emerged for both MDM2 and MDMX: LTFEHYWAQLTS, also termed pDI, and PMI–TSFAEYWNLLSP. The 3 most critical residues involved in p53-MDM2/MDMX recognition, i.e., Phe-19, Trp-23 and Leu-26 (p53 numbering), were all present in the phage-selected consensus sequences. The most potent inhibitor (TSFAEYWNLLSP), termed PMI, bound to MDM2 and MDMX at low nanomolar affinities—approximately 2 orders of magnitude stronger than the wild-type p53 peptide of the same length (ETFSDLWKLLPE). Comparative structural analysis identified an extensive, tightened intramolecular H-bonding network in bound PMI that contributed to its conformational stability, thus enhanced binding to the 2 oncogenic proteins. Importantly, the C-terminal residue Pro of PMI induced formation of a hydrophobic cleft in MDMX previously unseen in the structures of p53-bound MDM2 or MDMX. 1818 LTFEHYWAQLTS(7)[77][NT] TSFAEYWNLLSP(5)[4.2][8.9] DDWFQRVWSPLM(1)[NT][NT] NTFREYWNQLPT(1)[NT][NT] DDWFQRVVSPLM(1)[NT][NT] F-X(3)-W-X(2)-L 5 Phage display (common panning) 4 PMID:19255450 p53-binding domains of MDMX Cellular tumor antigen p53 3DAB, 3EQY,3FDO,3JZO,3JZP,3JZQ, Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR),Isothermal titration calorimetry (ITC) A surface plasmon resonance (SPR)-based competition assay was performed with (15–29)p53 as the competitor. The Kd value (nM) of synthesized peptides was shown. Besides, the direct interaction between p53-binding domains of MDM2 (referred to thereafter as (syn)MDM2) and the peptide (TSFAEYWNLLSP) were quantified at 25°C, using a MicroCal VP-ITC microcalorimeter. The binding affinity (Kd, nM) was shown. NT denotes not tested. The p53-binding domains of MDMX (referred to thereafter as (syn)MDMX) and their site-specifically biotinylated forms were chemically synthesized using native chemical ligation. In fact, biotin-(syn)MDMX was used as bait against the phage display library. The basic procedures for library screening are as follows: (i) incubate input phage (10 μL) with 400 μL of 10 nM biotin (syn)MDMX for 60 min before adding phage-target solution to 50 μL of streptavidin-agarose resin (Pierce) for affinity capturing; (ii) wash unbound phage; (iii) elute bound phage with 1 mM (15–29) p53; (iv) amplify the eluate and collect phage for the next round of panning; (v) repeat steps 1– 4; (vi) sequence selected binding clones according to the procedures recommended by the manufacturer. Two consensus sequences emerged for both MDM2 and MDMX: LTFEHYWAQLTS, also termed pDI, and PMI–TSFAEYWNLLSP. The 3 most critical residues involved in p53-MDM2/MDMX recognition, i.e., Phe-19, Trp-23 and Leu-26 (p53 numbering), were all present in the phage-selected consensus sequences. The most potent inhibitor (TSFAEYWNLLSP), termed PMI, bound to MDM2 and MDMX at low nanomolar affinities—approximately 2 orders of magnitude stronger than the wild-type p53 peptide of the same length (ETFSDLWKLLPE). Comparative structural analysis identified an extensive, tightened intramolecular H-bonding network in bound PMI that contributed to its conformational stability, thus enhanced binding to the 2 oncogenic proteins. Importantly, the C-terminal residue Pro of PMI induced formation of a hydrophobic cleft in MDMX previously unseen in the structures of p53-bound MDM2 or MDMX. Several MDMX structures have been determined in complexes with PMI (PDB ID code 3EQY) and pDI (PDB ID code 3FDO, 3JZO, 3JZP and 3JZQ). 1819 VDWVGWGASW(5) WVEWRRGWLS(1) SRTWVLWIRY(4) WGYVFHGWNL(3) MWGMHWAGEW(2) CCWGYGGSLV(1) 6 Phage display (common panning) 3 PMID:9048419 Polystyrene/polyurethane magnetic beads Not determined. Not determined. Not determined. fUSE5-based X10 phage display library Twenty randomly chosen clones from the third round of selection were analyzed further. Only clones binding significantly better to the M-280 beads than control phages from the unselected library or a wild-type phage without insert are shown. 1820 PWNAWGRTWV(5) VYRVIWFVGR(1) GGWAMWGRVW(2) FHAFHAWSAA(1) RPVAGDHPGS(1) GSVWWQAHVDR(1) WRLGFARWIG(3) WMRWGRSWGR(1) GSWKEWGGAW(1) LNWRGGWNWF(1) 10 Phage display (subtractive panning) 3 PMID:9048419 Polystyrene/polyurethane magnetic beads Not determined. Not determined. Not determined. fUSE5-based X10 phage display library A 15-mer synthetic peptide ODIN (GREPRVATVTRILRQ) was covalently attached to tosyl-activated M-280 beads by its N-terminal amino group. For counter selection, amplified phage from the first round of selection were incubated with beads coated no peptides. The supernatants were used as input to the second round. The initial rational was to apply the phage display technique to select peptides attached to magnetic beads. Individual phage clones, however, did not bind to ODIN-peptide adsorbed to microplate wells, where they bound strongly to uncoated as well as peptide-coated beads. Thus, the selection process had apparently resulted in enrichment of phages with affinity for the bead material and not for the peptide, despite deliberate attempts to eliminate plastic-binding clones from the phage pool prior to the second round of panning by counter-selection against beads without attached peptides. Twenty randomly chosen clones from the third round of selection were analyzed further. Only clones binding significantly better to the M-280 beads than control phages from the unselected library or a wild-type phage without insert are shown. 1821 SAWVRWGRVW(18) FLPYARWFSR(1) WSAWVGDWAL(6) VIYYFLVMFM(1) SRGFWLVWEQ(1) 5 Phage display (subtractive panning) 3 PMID:9048419 Polystyrene/polyurethane magnetic beads Not determined. Not determined. Not determined. fUSE5-based X10 phage display library A 15-mer synthetic peptide ODIN (GREPRVATVTRILRQ) was covalently attached to tosyl-activated M-280 beads by its N-terminal amino group. For counter selection, amplified phage from the first round of selection were incubated with beads coated no peptides. The supernatants were used as input to the second round. The initial rational was to apply the phage display technique to select peptides attached to magnetic beads. Individual phage clones, however, did not bind to ODIN-peptide adsorbed to microplate wells, where they bound strongly to uncoated as well as peptide-coated beads. Thus, the selection process had apparently resulted in enrichment of phages with affinity for the bead material and not for the peptide, despite deliberate attempts to eliminate plastic-binding clones from the phage pool prior to the second round of panning by counter-selection against beads without attached peptides. Twenty randomly chosen clones from the third round of selection were analyzed further. Only clones binding significantly better to the M-280 beads than control phages from the unselected library or a wild-type phage without insert are shown. 1822 VCFVMFSMCR(1) GVIHWRAWGGDW(1) LWGGLYGIWM(1) TWTLWFASMS(1) LSAWARWGSS(1) 5 Phage display (subtractive panning) 3 PMID:9048419 Polystyrene/polyurethane magnetic beads Not determined. Not determined. Not determined. fUSE5-based X10 phage display library A 15-mer synthetic peptide ODIN (GREPRVATVTRILRQ) was covalently attached to tosyl-activated M-280 beads by its N-terminal amino group. For counter selection, amplified phage from the first round of selection were incubated with beads coated no peptides. The supernatants were used as input to the second round. The initial rational was to apply the phage display technique to select peptides attached to magnetic beads. Individual phage clones, however, did not bind to ODIN-peptide adsorbed to microplate wells, where they bound strongly to uncoated as well as peptide-coated beads. Thus, the selection process had apparently resulted in enrichment of phages with affinity for the bead material and not for the peptide, despite deliberate attempts to eliminate plastic-binding clones from the phage pool prior to the second round of panning by counter-selection against beads without attached peptides. Twenty randomly chosen clones from the third round of selection were analyzed further. Only clones binding significantly better to the M-280 beads than control phages from the unselected library or a wild-type phage without insert are shown. 1823 PTWRSM(10) TRMRPG(1) VLLSVA(1) INQVRF(1) WCSRLF(1) PCHCSF(1) RGYFFK(1) 7 Phage display (common panning) 3 PMID:7682645 Monoclonal antibody 3-4B Not determined. Not determined. Not determined. f3-6mer phage display library (X6) ELISA The difference between A405 and A495 is taken as the signal. The strongest signals amount to ~0.6, while background is ~0.02. Only relatively strong ligands give signals in this assay. Phages displaying a ligand for the biotinylated antibodies are captured on the dish by binding of the biotin moiety to immobilized streptavidin. 1824 ATWAVL(1) AVMTSS(1) SWFLQW(1) REWISH(1) YTALLI(1) CYLCSV(1) LFSSGK(1) VWHLLH(1) VPWWVP(1) LSRILF(1) PGHSPW(1) WNLRSS(1) IALMDY(1) 13 Phage display (common panning) 3 PMID:7682645 Insulin Not determined. Not determined. Not determined. f3-6mer phage display library (X6) ELISA The difference between A405 and A495 is taken as the signal. The strongest signals amount to ~0.6, while background is ~0.02. Only relatively strong ligands give signals in this assay. Phages displaying a ligand for the biotinylated insulin are captured on the dish by binding of the biotin moiety to immobilized streptavidin. 1825 RAWSYV(1) KGRYQQ(1) EHGRPQ(1) GCSDVL(1) NLLSMT(1) CLGEHD(1) RFYGGS(1) ILPLRI(1) LSRSYF(1) NLYLVH(1) AWFRRL(1) LRGKLS(1) VDVGRS(1) 13 Phage display (common panning) 3 PMID:7682645 Interleukin-2 receptor subunit alpha Interleukin-2 (IL-2) 1Z92,2B5I,2ERJ, Not determined. f3-6mer phage display library (X6) ELISA The difference between A405 and A495 is taken as the signal. The strongest signals amount to ~0.6, while background is ~0.02. Only relatively strong ligands give signals in this assay. 1826 FHENWPS(15) 1 Phage display (common panning) 3 PMID:11839467 96-well immunoassay plate (polystyrene, PS) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The authors' purpose is to identify phage peptides that recognize the HER modification on BSA. A single sequence FHENWPS, emerged after the third round for all 15 independent clones, which appeared in one clone in the second biopanning round. Screening of the clone against an array of positive and negative targets to determine binding efficiency showed high readings (OD>1.5) in ELISA analysis. However, there were no differences between either positive versus negative target-coated or target-coated versus BSA-blocked uncoated wells by ELISA, nor by a more sensitive method of plaque assay. They concluded that most probably the selected clone was a super-binder of polystyrene surfaces that grew preferentially in the third biopanning round, and a different biopanning strategy for BSA-HER in solution was developed. 1827 FHWTWYW 1 Phage display (common panning) http://www.neb.com/nebecomm/tech_reference/protein_tools/phdFaq.asp#4.13 96-well immunoassay plate (polystyrene, PS) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The sequence FHWTWYW is a plastic binder discovered and characterized at New England BioLabs (NEB). 1828 APFYYSWFPSYMAEGLPSPSPLDPVQDNLY 1 Phage display (common panning) PMID:15620878 96-well immunoassay plate (polystyrene, PS) Not determined. Not determined. Not determined. Not determined. 1829 WHRWPWLVSG(1) WHWWYWALDR(1) WHWWXW 2 Phage display (common panning) 3 PMID:9237215 Petri dish (polystyrene, PS) Not determined. Not determined. Not determined. X10 phage display library Each selection was performed on the streptavidin-coated (Pierce, Rockford, IL) petri plates. The peptide sequences WHRWPWLVSG and WHWWYWALDR were isolated from streptavidin biopanning, each at a frequency of 1 of 43 phages sequenced. Most importantly, WHWWXW phage have been isolated repeatedly in biopannings in which no streptavidin was used. The amino acid composition and the presence of the WHXW motif suggest that their phage were also plastic binders. 1830 YKKPLLMDQILK GDIDSQGVRPHE IPDQGNKPKSRL THQKPSDLPIGH WTAASSVPSKSS LTPSVWSSKSHN AQVPYFSNHPSG FDPFLRTLQGTH 8 Phage display (common panning) 3 PMID:21600948 Mouse pooled sera immunized with VHBL plasmid DNA VHBL plasmid DNA Not determined. Not determined. Ph.D.-12 phage display library (X12) 1831 CLPWAQHMC CVSLPHRTC CPSPTRSAC CTKMSPHAC CFNRSPVVC CGIRRTPTC CPLSKFHVC CPLTLPATC CKQQPYLYC CTKCQKMAC CTKMSRHAC CTKCRNNAC CTTSLYRSC CNGFYATTC CWLGDITQC CHSWLIHNC 16 Phage display (common panning) 3 PMID:21600948 Mouse pooled sera immunized with VHBL plasmid DNA VHBL plasmid DNA Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 1832 SCQLEGSRLRCP QCKLEGSRLRCV QCALEGARIRCS QCTMDQGRLRCR GCITEQGRSRCI SCKLEGSRLRCP TCATDQGRLRCT HCFHDQGRVRCA TCEMTQGRLRCV ACRSDQGRARCT NCTLEGTRFRCS RCKPDQGRLRCG SCMKDPSSPRCL HCTMDQGRLRCR SCMLDQGRSRCR TCRQTLTPAVCH QCQEDQGRRRCS 17 Phage display (common panning) 3 PMID:21763377 Anti-mut-II monoclonal antibody LmmAbB2D4 Mutalysin-II (mut-II) Not determined. Not determined. XCX8CX+X15+X8CX8+X30 M13 phage display library pool The DNA sequence and the deduced amino acid sequence were determined, showing that the peptides were selected from the (XCX8CX) library. LmmAb2D4 reactivity with cellulose-bound phage-display identified peptides was detected with an alkaline-phosphatase coupled anti-mouse antibody. Spot intensities were measured with the Scion Image software. The results identified the QCTMDQGRLRCR, TCATDQGRLRCT, HCFHDQGRVRCA, HCTMDQGRLRCR and SCMLDQGRSRCR sequences as possible epitopes. 1833 CRNADKFPC CLDNQRPKC CDCRGDCFC 3 Phage display (in vivo) 5 PMID:21768392 Rat synovial tissue Not determined. Not determined. Not determined. CX7C fUSE5 phage display library Authors identified and isolated phages specific for arthritic joints using a combination of ex vivo and in vivo phage screening. For ex vivo screening, authors used CD31-expressing endothelial cells from the joints of an arthritic rat. Two rounds of ex vivo enrichment produced a phage pool that bound 66-fold more efficiently to the endothelial cells compared with the nonrecombinant phage. The ex vivo selected phage was injected i.v. into an arthritic rat, and subsequent three rounds of in vivo selection yielded a 53-fold enrichment of the phage recovered from the synovial tissue, whereas no enrichment was observed in the control tissues, namely the lung and the kidney. Apparently NQR and ADK bind to different receptors/receptor domains on the endothelial cell surface than RGD. We draw this inference based on three findings: (i) no cross-inhibition of phage binding by these peptides; (ii) inhibition of attachment of HUVECs to vitronectin, which involves integrin αvβ3, by RGD peptide, but not by NQR or ADK peptide; and (iii) differential alterations in the MAP kinase signaling pathway events induced by NQR peptide versus RGD peptide. The precise identity of the receptors that bind NQR and ADK peptides to vascular endothelial cells remains to be determined. In terms of functional properties, NQR and RGD, but not ADK, suppressed arthritis. ADK peptide apparently binds to a receptor that does not trigger a detectable tissue response. The antiarthritic activity of NQR is likely attributable, at least in part, to inhibition of angiogenesis and resulting inhibition of leukocyte migration into the inflamed joint. 1834 SSKVYTPTGSLP(1) TLSWGLGSIRTA(1) EDQYLSSEDFLL(1) VFPNNGLALSHV(1) FPGAAGSNRSPL(1) HDTSETKGLTDT(1) TYPNKYTAITPD(1) MKTAMPIKTKWP(1) LPKGSTDSGNMK(1) TLSWGLGSIRTA(1) AEYSMYSAHRPR(1) WYPNKYSPFHQS(1) AQPPKSEGWTLR(1) ATEIIPIALSRP(1) AQPPKSEGWTLR(1) SWLQLVTSKSTL(1) TNSILFARAPAE(1) GYFPLRLPLLSL(1) 18 Phage display (common panning) 3 PMID:21781322 Anti-CAEV polyclonal antibody Caprine arthritis encephalitis virus (strain 63) (CAEV-63) Not determined. Not determined. Ph.D.-12 phage display library (X12) 1835 CTKLSHKNC(2) CHTALAPSC(1) CGTLINNQC(2) CGWHSPMYC(1) CAVFYTTNC(1) CYNRAMIGC(1) CNFFTEPLC(1) CSGKFYPVC(1) CNLPSDPFC(1) CPSHDPWVC(1) CLLQSDPFC(1) CLSKDPFVC(1) CNSDPWVC(1) CSWNHWSYC(3) CSWKHWSYC(1) L-x-SDP-[FY] and SW-[NK]-HWSY 18 Phage display (common panning) 3 PMID:21781322 Anti-CAEV polyclonal antibody Caprine arthritis encephalitis virus (strain 63) (CAEV-63) Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Authors obtained families of 7 mer constrained peptides with consensus motifs LxSDPF/Y and SWN/KHWSY and mapped the epitopes mimicked by them at the Env 6-LISDPY-11 and 67-WNTYHW-72 sites of the mature gp135 amino acid sequence. The first epitope fell into the N-terminal immunogenic aa1-EDYTLISDPYGFS-aa14 site identified previously with a synthetic peptide approach; the second epitope has not been described previously. The first epitope is mostly conserved across CAEV isolates whereas the second newly described epitope is extremely conserved in Small Ruminant Lentiviruses env sequences. As being immunodominant, the epitopes are candidate targets for mimotope-mediated diagnosis and/or neutralization. 1836 CRTLTVRKC CIKRGGKRC 2 Phage display (subtractive panning) 4 PMID:21781994 Recombinant 4e5 domain of human stabilin-2 Not determined. Not determined. Not determined. CX7C T7 phage display library For subtraction of non-specific binding, the 1.0e9 plaque-forming units (pfu) in 100????l of TBST (50 mM Trisa??a After 46 selected phage obtained in the second-to-fourth rounds were sequenced, authors identified the two most commonly occurring clones, with peptide sequences CRTLTVRKC and CIKRGGKLC. ClusterX alignment analysis of all identified peptide sequences did not reveal any shared motif. To confirm selective binding of the two phage clones to the 4e5 domain, a binding assay was conducted. The two clones bound strongly to the human stabilin-2_4e5 protein, compared with insertless phage, and did not bind to other control proteins including the βig-h3 FAS domain, bovine serum albumin, or collagen, suggesting that the two clones bound specifically to the 4e5 domain. Authors examined whether selective binding of the CRTLTVRKC- or CIKRGGKLC-expressing phage to the human stabilin-2_4e5 domain was mediated by the peptides displayed by the phage. To this end, FITC-conjugated synthetic peptides were prepared. Fluorescence microscopic examination indicated that binding of the CRTLTVRKC (S2P) peptide to L/Stabilin-2 cells was significantly higher than to L/MOCK cells. However, the CIKRGGKRC peptide did not bind to either L/Stabilin-2 or L/MOCK cells, as was also the case with NSSSVDK, used as a control peptide. The peptide-binding to L/Stabilin-2 or L/MOCK cells was confirmed via FACS analysis. 1837 CTSTSAPYC(5) CVCCLAGLC(1) CNKWNLLNC(1) CTHDYPLVC(1) CPFGSPALC(1) CMRTHHMQC(1) CWANQPATC(1) CMTRHHLDC(1) 8 Phage display (in vivo) 4 PMID:21792566 ICR male mice brain parenchyma Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Following sequential rounds of isolation, a number of phages were sequenced and a peptide sequence (CTSTSAPYC, denoted as PepC7) was identified. Clone 7-1, which encodes PepC7, exhibited translocation efficiency about 41-fold higher than the random library phage. Immunofluorescence analysis revealed that Clone 7-1 had a significant superiority on transport efficiency into the brain compared with native M13 phage. Clone 7-1 was inhibited from homing to the brain in a dose-dependent fashion when cyclic peptides of the same sequence were present in a competition assay. Interestingly, the linear peptide (ATSTSAPYA, Pep7) and a scrambled control peptide PepSC7 (CSPATSYTC) did not compete with the phage at the same tested concentration (0.2-200 pg). Labeled by Cy5.5, PepC7 exhibited significant brain-targeting capability in in vivo optical imaging analysis. The cyclic conformation of PepC7 formed by disulfide bond, and the correct structure itself play a critical role in maintaining the selectivity and affinity for the brain. In conclusion, PepC7 is a promising brain-target motif never been reported before and it could be applied to targeted drug delivery into the brain. 1838 LPWWLPYRGESN(3) SPLSWWPHATVG(2) YGPWWYSSNAES(2) LPWWPQASISPP(1) LPWWPIQRVSHL(1) LPWWIPKEGWAV(1) LPWWLPPSLSRV(1) AGPWWHQTSVHV(1) NIPWWPFSLHAP(1) GGAWWPTSLVMY(1) APYSWWPYSAYN(1) SSVKSWWPAFTP(1) WYPQPFWPYRQA(1) YGKPFWPSSLWW(1) IPYWPFLPDTSM(1) LPYWLPYSSGNK(1) VPYWMPPPTVIP(1) PWWP 17 Phage display (common panning) 5 PMID:21816124 Plasmodium berghei ookinetes Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Fluorescence Specific peripheral immunofluorescence was uniformly observed on ookinetes incubated with selected phage clones 5 (LPWWPIQRVSHL) and 14 (SPLSWWPHATVG). Ookinetes incubated with non-selected phage clone and/or incubated with secondary antibody only showed no signal. The 21 selected phage peptides had adjacent residues of tryptophan (W) and proline (P) within their sequence, with proline residues flanking tryptophan. Prolines (8/21) were frequently associated with leucine (L) residues. A consensus peptide sequence (PWWP) was identified in phages that bound to the Plasmodium berghei ookinete surface and, in selected phages, bound to actin and enolase in overlay assays with ookinete protein extracts. Actin was localized on the surface of fresh live ookinetes by immunofluorescence and electron microscopy using specific antibodies. The overall results indicated that enolase and actin can be located on the surface of ookinetes, and suggest that they could participate in Plasmodium invasion of the mosquito midgut. 1839 CVKWRGVVVC 1 Phage display (common panning) 4 PMID:21821708 Membrane primary amine oxidase Sialic acid-binding Ig-like lectin 9 (Siglec-9) Not determined. Not determined. CX8C phage display library After 4 rounds of panning using recombinant VAP-1 as a bait, we obtained a 400-fold enrichment of phages bound to VAP-1 compared with the control (BSA-coated wells). Of the 23 randomly selected phages sequenced, 19 gave a sequence. Database searches with the sequence derived from the phage clones revealed similarities to the amino acid sequence of Siglec-9 (residues 289-295). In the binding assays, the phage peptide bound selectively to recombinant VAP-1. The binding between Siglec-9 and VAP-1 was confirmed by in vitro and ex vivo adhesion assays. The interaction sites between VAP-1 and Siglec-9 were identified by molecular modeling and confirmed by further binding assays with mutated proteins. 1840 RGR RQR RRY TRR RPR RRA QRR RRG 8 Phage display (common panning) 5 PMID:21821948 Soluble 37/48kDa oligomer formation of Aβ(1-42) Not determined. Not determined. Not determined. X3 T7 phage display library After the fifth round, arginine-containing peptides were enriched in phage library. 1841 RRRA RGKK KAVR RFRK RRRL RSKK RRKV KRAS 8 Phage display (common panning) 5 PMID:21821948 Soluble 37/48kDa oligomer formation of Aβ(1-42) Not determined. Not determined. Not determined. X4 T7 phage display library After the fifth round, arginine-containing peptides were enriched in phage library. SDS-PAGE and size-exclusion chromatography indicated that the inhibitory activities of 4-residue peptides against the soluble 37/48kDa oligomers of Aβ42 were higher than those of the 3-residue peptides, and RFRK exhibited strong inhibitory activity as well as SRPGLRR. SEPGLRR, a 7-residue peptide found by phage display, exhibited inhibitory activity against soluble 37/48kDa oligomers of Aβ42 (Mimotope Set ID: 1317) 1842 CLRSHPLGC CTFASVMTC 2 Phage display (common panning) PMID:21829538 Soluble ectodomain of APP isoform sAPPα695 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Five out of 28 individually picked sAPPa-binding clones contained the peptide sequence LRSHPLG. An additional sAPPa-binding clone comprising amino acid sequence TFASVMT was isolated. The sequences of both peptides were then compared to human protein sequences in the NCBI databank using BLAST. This analysis revealed that both peptides presented significant sequence homologies to several members of the human semaphorin protein family. Interestingly, the TFASVMT peptide is homologous to a region of semaphorin 3A that has been shown to be important for receptor recognition, while the region that is homologous to the LRSHPLG peptide lies very close to that same important site. These initial results suggested that sAPPa binds to semaphorins at a domain that is important for receptor recognition and biological activity. 1843 CLLGTRWPC CFMESVGRC CLTPEFHIC CLAVGEVLC CFVSPPVGC CMPGWEVLC CKRGNSGSC CQRLVGFAC CLQASPNFC CNGSVRSFC CPGFGLAYC CFLFTFEAC CRGVLMRYC CRAFVVASC CQSHSAFVC 15 Phage display (subtractive panning) 4 PMID:21829599 Human thymic CD4+CD25+ T cells Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method, which allows the separation of cell-bound from unbound phage in one step. CD4+CD25- cells were used for preclearing. Among the phages sequenced a specific phage peptide for further analysis due to its sequence similarity to the Vitamin D receptor (VDR). This was the first choice because although Vitamin D has been extensively reported to display immunoregulatory functions in different contexts, its role has not been well-defined for human thymic T regs, opening an interesting area for research. The results from the ELISA assay showing that VDR phage was able to bind to active Vitamin D strongly supports our hypothesis. When phages were preincubated with Vitamin D we did not observe binding to VDR, confirming that the peptide, in fact, mimic the Vitamin D binding pocket in VDR and is able to prevent Vitamin D from binding to its native receptor. 1844 HWGNHSKSHPQR(3) HTLHRQVPKHWL(2) HYQHNTHHPSRW(2) HSSSASDRSRPL(1) STHHRHYHDTLA(1) GHIHSMRHHRPT(2) KHMHWHPPALNT(28) 7 Phage display (common panning) 5 PMID:21856287 PreS1 protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Alignment of the positive phage clones revealed a putative consensus PreS1 binding motif of H-(X)n-H-(X)m-H-P/R. Moreover, a peptide named P7 (KHMHWHPPALNT) was highly enriched and occurred with a surprisingly high frequency of 72%. A thermodynamic study revealed that P7 has a higher binding affinity to PreS1 than the other peptides. Furthermore, P7 was able to abrogate the binding of HBV virions to the PreS1 antibody, suggesting that P7 covers key functional sites on the native PreS1 protein. This newly isolated peptide may, therefore, be a new therapeutic candidate for the treatment of HBV. 1845 QDEERVSSCPKVAWTFC NCNKNDHLFACW ECKSDWMPPYCP ECTQKYDWLFCM 4 Phage display (common panning) 3 PMID:21872636 Anti-Li venom monoclonal antibody LimAb7 Sphingomyelin phosphodiesterase D LiSicTox-alphaIA1bi Not determined. Not determined. LX-8, X15, X8CX8, X30 phage display library pool These mimotopes, together with a 3D model of LiD1, were used to predict with the MIMOP bioinformatics tool the putative epitope region (residues C197, Y224, W225, T226, D228, K229, R230, T232 and Y248 of LiD1) recognized by LimAb7. This analysis and results of alanine-scanning experiments highlighted a few residues (such as W225 and D228) that are found in the active site of different SMases D and that may be important for LiD1 enzymatic activity. 1846 HAWDPIPARDPF(4) AAWHLIVALAPN(1) ATSHLHVRLPSK(5) 3 Phage display (subtractive panning) 5 PMID:21887228 Influenza A virus (A/goose/Jilin/hb/2003(H5N1)) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Since the viruses were harvested from the cells, therefore, a subtract panning was conducted. In the fourth round of panning, the coated viruses were replaced by the supernatant form Madin-Darby canine kidney (MDCK) cells culture. Putative binding motifs to H5N1 viruses were identified by DNA sequencing. In terms of the minimum quantity of viruses, the phage-based ELISA was better than antiserum-based ELISA and a manual, semi-quantitative endpoint RT-PCR for detecting H5N1 viruses. More importantly, the selected phages bearing the specific peptides to H5N1 viruses were capable of differentiating this virus from other avian viruses in enzyme-linked immunosorbent assays. 1847 CTGSTQHQC CHSALTKHC CSTHFIDTC 3 Phage display (common panning) 5 PMID:21903933 Porcine skin Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Screening studies were performed on porcine skin in Franz diffusion cells (FDCs, PermeGear). The phage display library was placed in the donor compartment of the FDC. After 24 h, phage which had penetrated through the skin were collected from the receiver compartment, amplified, and again placed in the donor compartment for an additional round of screening to further narrow down the library. Peptides that penetrate the SC were identified using in vitro phage display, which were labeled a skin permeating and cell entering (SPACE) peptide. In vitro studies indicated that the SPACE peptide, when conjugated to cargoes such as small molecules and proteins, was able to facilitate their penetration across the SC into epidermis and dermis. 1848 CKTGSHNQC(0.3) CMGPSSMLC(0.3) CTDPNQLQC(0.2) 3 Phage display (common panning) 5 PMID:21903933 Porcine dermis Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) For the dermis screen, the phage display library was also placed in the donor compartment of the FDC. After 24 h, the liquid from the donor compartment was removed and the skin was placed at 60 °C for 90 s. The epidermis was then removed from the dermis. To extract phage from the dermis, the dermis was cut up into small pieces and then homogenized (IKA disperser). Phage screening was performed to isolate phage that localized in the dermis. 1849 CDPSPHTHC(7) CPSSLTPAC(2) CFPHGLGLC(1) CSPAVPMLC(1) 4 Phage display (common panning) 3 PMID:21906439 Ig-like C2-type 1-3 domain of c-kit Kit ligand 2E9W, Not determined. Ph.D.-C7C phage display library (CX7C) The synthesized peptides CDPSPHTHC and CPSSLTPAC, particularly CPSSLTPAC, stimulated UT-7 cell proliferation. 1850 HEPSSFVFFPLS(1) NALKPQGLHSWY(1) AVSSFERDNFVQ(2) TPLKAHLHSQRH(1) NSFHVPRLLIAG(1) THSFIPRLPLLK(1) WPKYTPRVFPHF(1) 7 Phage display (common panning) 3 PMID:21906439 Ig-like C2-type 1-3 domain of c-kit Kit ligand 2E9W, Not determined. Ph.D.-12 phage display library (X12) The synthesized peptides AVSSFERDNFVQ and THSFIPRLPLLK, particularly THSFIPRLPLLK, stimulated UT-7 cell proliferation. 1851 TMRGKKKRTRAN(1) QHRMASMSPTLP(1) GLLPYRPREANF(1) AMIPYTWFSPSP(1) KQPKKAPRRIPQ(1) SIPTTWFHPPPS(1) GVSLHNTNWNIY(1) SDTSVNWLTLWY(1) NTPQRPPYKRSP(1) LAKSPSNSAREW(1) AKCHSDVPSPAC(1) VHFKPTHLPSPP(1) STSQALSRFPSF(1) GMMRALSHPSAS(1) GTLTTPRLDLIM(1) MKISAPALAFGL(1) MFAKSPPYPSLM(1) FNWHWLSRPYFP(1) FANHLTNAVHAL(1) SQPWTNALVVSS(1) KLWNVPWPPHMR(1) FTPPPAYGRNEG(1) TAFWPLYPLSDW(1) HWIPQTLPASFI(1) HHPFVTNTPSLI(1) PNRLGRRPVRWE(1) HWWYPLLPVRQM(1) 27 Phage display (common panning) 3 PMID:21926473 EWS-Fli1 protein RNA helicase A, RHA Not determined. Not determined. Ph.D.-12 phage display library (X12) The cytotoxicity evaluation of these peptides with in EWS-FLI1 containing cell lines yielded one potent peptide called ESAP1 (TMRGKKKRTRAN). ESAP1 binds EWS-FLI1 with 0.202 micromolar affinity as measured in surface plasmon resonance. The minimal interaction region of ESAP1 is characterized and found that the lysine residues are critical for cellular cytotoxicity. ESAP1 reduces the transcriptional activity of EWS-FLI1 as well as disrupts cell cycle kinetics in ewing tumor cells. these findings provide both a novel experimental probe and a potential therapeutic scaffol d for ewing tumor. 1852 GAMHLPWHMGTL 1 Phage display (subtractive panning) 3 PMID:21928455 Zinc oxide, ZnO Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phages were first incubated with the zinc precursor (zinc nitrate solution) at room temperature. Careful selection of the Zn precursors is important for this method. If conventional Zn precursors that form zinc hydroxide solids in aqueous solution are used, amorphous Zn(OH)x might grow in addition to ZnO nanoparticles in the phage solution, and then the panning process could also contain the sequences of peptides that bind amorphous Zn(OH)x nanoparticles. The results suggest that the GAMHLPWHMGTL peptide can not only nucleate the catalytic growth of semiconductor nanoparticles, but can also induce the anisotropic coagulation of primitive crystalline domains at room temperature to form single-crystalline nanoparticles easily and without the need to add complex capping reagents. 1853 DRYSEIRTSSTL(6) YSGLQDSSLRLR(4) DYPSANKWPRYV(1) 3 Phage display (subtractive panning) 3 PMID:21930161 The tegument of schistosoma japonicum schistosomula Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Western blot To confirm the binding ability of positive phages, the MppZL1 (DYPSANKWPRYV), MppZL4 (YSGLQDSSLRLR), MppZL6 (DRYSEIRTSSTL) and negative control M13KE phage were tested for recovery rate. Significantly higher binding was observed by MppZL4 to schistosomula than by MppZL6 or MppZL1 or control M13KE phage. The results of Western blot indicated that the peptide of the MppZL4 exhibited strong affinity for the surface membrane or tegument of schistosomula. Cercariae was directly used for construction of a reverse adsorption peptide library. Of the three, M13 phage peptide ZL4 (MppZL4, YSGLQDSSLRLR, 1.4 kDa, pI 8.8) bound to the tegument of mechanically transformed schistosomula and to other developmental stages of S. japonicum from the mammalian host. By contrast, MppZL4 did not bind to the surface of cercariae. 1854 DFSVLQTIGDSL(0.4) 1 Phage display (competitive panning) 3 PMID:21953453 BH3 motif of Mcl-1 protein 26-amino acid Bim peptide Not determined. Not determined. Ph.D.-12 phage display library (X12) 1855 DFSVLQTIGDSL(0.2) NETVNTMLTYYY(0.2) NETVELMQAYLH(0.2) 3 Phage display (competitive panning) 3 PMID:21953453 BH3 motif of Mcl-1 protein Sabutoclax Not determined. Not determined. Ph.D.-12 phage display library (X12) 1856 CGTFAHPQC(1) CGTFIHPQC(4) 2 Phage display (common panning) 3 PMID:16258189 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. Ph.D.-C7C phage display library (CX7C) 1857 CGTFAHPQC(5) CGTFDHPQC(3) 2 Phage display (common panning) 3 PMID:16258189 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. Ph.D.-C7C phage display library (CX7C) 1858 ACTTHPQNKPKEQRCL AADKWSHPQNPLRSPTMA AADKWSHPQAPVPLLRPA YTTAHPQGDLSV HCMSAHPQDLCT 5 Phage display (common panning) PMID:15620878 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. Not determined. 1859 HGMASQYFTCFHDSEPSSPGMFGWDPTTPTLPHPQVDE IAHRVVAYNSLDSNPIWLPGEESSSVFGDYHPMFRAPV HVPVFTRYNYAKPNDTDWPGGFVDSLSAHPQGPIAGGR MTLGYDRASPAPNTSFSNPGLDFNPFTYHPQGPHQILQ AGRAARDDDCRGHACMIIPGVSLFNSDHPMGAHPSIRR DFSSFLTGTNAMAPFWPFPGSTYLLGHPMAPRDLQTSN SASWKFNSSFGYPTGGIEPGPNCHPQACPDVALKSLSP VSEMSSFSGCNTDHHPQGPGGRHDIMRSISESRGYGSL EMLTLPLTSIPIPWHPQGPGYLYHKPPRGTDFRMLSSK PYRFYHPYSHPRHPQGDVPGSSAEVFHTFPNTQGRNSR ADYGTIGESPCHPQVDICPGALHHEFNEFFVGMSPEPS ARMAGLTEHPQGDIIDHHPGWVHDSKISPRNQDTYHSS AHLFGHPQVGFDSIGSAFPGDIHCKQYKADSGLQSAAA PDYDLMSSTCRFYGCSKMPGGVAVNGLFAVQGHSKYSS TWDFTRSSLPAGDTSFTFPGSYSVMTRSCGISCVPAEV H-P-[QM] 15 Phage display (common panning) 3 PMID:8508960 Streptavidin Biotin 1DF8,1LUQ,1MEP,1MK5,1N43,1N9M,1NDJ,1NQM,1STP,1SWD,1SWE,1SWG,1SWK,1SWN,1SWP,1SWR,1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,3MG5, Not determined. TSAR-9 phage display library (X18PGX18) SA selected two classes of phage from the library. One class exhibited the aa motif, HP(Q/M)Θ (where Θ signifies a non-polar aa), similar to the motif identified by Devlin et al. [Science 249 (1990) 404-4061 using a 15-aa random peptide library displayed on phage. The other class of phage had no discernible motif. In binding experiments, the non-HP(Q/M)Θ phage had a slightly higher affinity for SA than did the motif phage. Both classes of SA-binding phage failed to bind native and non-glycosylated forms of avidin, even though SA and avidin are structurally similar and both proteins possess extraordinary affinities for biotin. 1860 DVDMGTIFNTIANNITSRPGVSWGGSTRTITKPKGAVA AQTAGQPGRTLSKPPIPNTPGPREPSLLHSMPHLPNLTA VRTISKPVAREGWTRDTVPGPATSIVEKRFHLIGVNAQ KGASFYPQCGGECQIYRVPGDHLPLFSLHRTGTPRHDS 4 Phage display (common panning) 3 PMID:8508960 Goat anti-mouse IgG (Fc) polycloanl antibody Mouse IgG Fc Not determined. Not determined. TSAR-9 phage display library (X18PGX18) The polyclonal goat anti-mouse IgG Fc Ab preparation selected phage displaying sequences similar to a region of the mouse IgG Fc. Thus, a single immunodominant epitope on the mouse IgG Fc was identified. 1861 CRLALGDAKKYC[0.180 ± 0.001] CVRKGGLIKGRC[0.153 ± 0.002] CGPRDRGGLIKC[0.130 ± 0.002] CDSNRGGLWRKC[0.158 ± 0.010] DRGGL 4 Phage display (common panning) 4 PMID:17975163 Anti-CEA monoclonal antibody Col-1 Carcinoembryonic antigen-related cell adhesion molecule 5 Not determined. Not determined. CL10 phage display library (CX10C) ELISA In ELISA, phage clones were bound by coated anti-CEA antibody Col-1 and detected by a peroxidase-labeled rabbit anti-phage antibody to determine specificity of peptides. Absorbances at 450-630 nm were determined, reproduced from the graph and shown as means ± standard deviations. The absorbance for negative control was 0.020. No phage binding occurred to isotype control antibody. According to the results of phage ELISA, all of the four independent phage clones bearing above peptides were specifically bound by the antibody Col-1 but were not recognized by an isotype control. No reactivity was observed with the wild-type phage. The competitive ELISA evidenced that the selected mimotopes were true mimics of the Col-1 epitope on the CEA antigen and clone bearing CRLALGDAKKYC peptide was the best circular mimotope candidate. 1862 DRGGLMKTN[0.169 ± 0.003] DMGGLFRKG[0.288 ± 0.004] DRGGLWKTP[0.141 ± 0.005] DRGGL 3 Phage display (common panning) 4 PMID:17975163 Anti-CEA monoclonal antibody Col-1 Carcinoembryonic antigen-related cell adhesion molecule 5 Not determined. Not determined. LL9 phage display library (X9) ELISA In ELISA, phage clones were bound by coated anti-CEA antibody Col-1 and detected by a peroxidase-labeled rabbit anti-phage antibody to determine specificity of peptides. Absorbances at 450-630 nm were determined, reproduced from the graph and shown as means ± standard deviations. The absorbance for negative control was 0.020. No phage binding occurred to isotype control antibody. According to the results of phage ELISA, all of the four independent phage clones bearing above peptides were specifically bound by the antibody Col-1 but were not recognized by an isotype control. No reactivity was observed with the wild-type phage. The competitive ELISA evidenced that the selected mimotopes were true mimics of the Col-1 epitope on the CEA antigen and clone bearing CRLALGDAKKYC peptide was the best circular mimotope candidate. 1863 RGGRCLLCCLCLWWA AVAGGRSVVDARVAR RTEVPVLSFTSPLTG PFARAPVEHHDVVGL RVPPRYHAKISPMVK 5 Phage display (subtractive panning, in vivo) 3 PMID:20692225 Human DA-MB-435eb.1 tumor xenografts Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Before positive selections, the library was subjected to negative selection by passage through non-tumor-bearing mice. Following in vivo selection from the f3-15mer phage display library in mice harboring tumor xenografts, two phage clones (RGGRCLLCCLCLWWA and AVAGGRSVVDARVAR) predominated in final outputs. These clones had no discernible affinity for the tumor cell line in vivo or in vitro but had unusually high infectivities, prompting further investigation. 1864 CWWLHPTHC(17) CPRTHYAIC(3) CAVCTDFC(1) CHLFTQAFC(1) CALFKPSMC(1) CQTSAMRHC(1) CSLNPSSRC(1) CYFNQPIRC(1) CLTLNGSPC(1) CSSKTSFTC(1) 10 Phage display (common panning) 4 http://acta.chem-soc.si/56/56-3-712.html Phospholipase A2, ammodytoxin C Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The interaction of selected peptides with pancreatic phospholipase A2 and ammodytoxin C was confirmed with surface plasmon resonance and phage ELISA assays. Interestingly, peptides showed equal affinity to both proteins, regardless which of the two proteins was used as a target in the selection procedure. 1865 HWWSPWPNPPQI(3) HWNVWWPVSIPE(1) HSWSFFWQSPAD(1) HSYWYRWTPSHL(1) FHWRWSTFPEYP(1) HWRWWQSDHLFT(1) SHWIRYFPWSIG(1) 7 Phage display (common panning) 4 http://acta.chem-soc.si/56/56-3-712.html Phospholipase A2, ammodytoxin C Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The interaction of selected peptides with pancreatic phospholipase A2 and ammodytoxin C was confirmed with surface plasmon resonance and phage ELISA assays. Interestingly, peptides showed equal affinity to both proteins, regardless which of the two proteins was used as a target in the selection procedure. 1866 CHSNRLNLC(3) CNGMYAHPC(3) CNMTQAYSC(1) CYRQASDSC(1) CQLQPTRLC(1) CDHHTYNHC(1) CHSPTRGIC(1) CKDSSLHVC(1) 8 Phage display (common panning) 4 http://acta.chem-soc.si/56/56-3-712.html Pancreatic phospholipase A2 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The interaction of selected peptides with pancreatic phospholipase A2 and ammodytoxin C was confirmed with surface plasmon resonance and phage ELISA assays. Interestingly, peptides showed equal affinity to both proteins, regardless which of the two proteins was used as a target in the selection procedure. 1867 KLWNLHPTQALW(2) APWHLSSQYSRT(2) GPSVGVTASHTR(1) SNHPATLTGTGG(1) AQPYPFSTRHWQ(1) 5 Phage display (common panning) 4 http://acta.chem-soc.si/56/56-3-712.html Pancreatic phospholipase A2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The interaction of selected peptides with pancreatic phospholipase A2 and ammodytoxin C was confirmed with surface plasmon resonance and phage ELISA assays. Interestingly, peptides showed equal affinity to both proteins, regardless which of the two proteins was used as a target in the selection procedure. 1868 CLFHHQASC(2) CTARSPLLC(2) CQVPRSPYC(1) CTTRTMTQC(1) CSMSTTRSC(1) CLETASNYC(1) CTSAPHRMC(1) CLPSRSHLC(1) CFWQSDKIC(1) 9 Phage display (competitive panning) 4 http://acta.chem-soc.si/56/56-3-712.html Pancreatic phospholipase A2 MJ33 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The interaction of selected peptides with pancreatic phospholipase A2 and ammodytoxin C was confirmed with surface plasmon resonance and phage ELISA assays. Interestingly, peptides showed equal affinity to both proteins, regardless which of the two proteins was used as a target in the selection procedure. 1869 HGWLYPHPRYPV(16) HLWPFYSMPPQH(2) HPPYWYPWQQSS(1) 3 Phage display (competitive panning) 4 http://acta.chem-soc.si/56/56-3-712.html Pancreatic phospholipase A2 MJ33 Not determined. Not determined. Ph.D.-12 phage display library (X12) The interaction of selected peptides with pancreatic phospholipase A2 and ammodytoxin C was confirmed with surface plasmon resonance and phage ELISA assays. Interestingly, peptides showed equal affinity to both proteins, regardless which of the two proteins was used as a target in the selection procedure. 1870 YAPWNIPLHMYL(3) FHKNWPIWWYPP(3) KLGDPKIATTLF(1) IPHPKLGDPRPM(1) SVTKLGDPRAPR(1) EQMSWMQLLVEM(1) SSRTKLIYQAAS(1) EQMSVAQFTRA(1) STSNYASASSPN(1) NNLIAAFLRTPI(1) FHKNWPIWWYPP(1) WHWYWQPAARGN(1) NPLHAASTIWWF(1) IVLPLSAPTALK(1) TRAAVLISSTWI(1) THRLDNSYSRMS(1) K-[LPV]-G-D-P-[RKL] 16 Phage display (common panning) 3 PMID:21764227 Rabbit anti-PrV polyclonal antibody IgG Pseudorabies virus, PrV Not determined. Not determined. Ph.D.-12 phage display library (X12) Groups of six C57BL/6 mice were immunized with bacteriophages expressing peptides with this motif sequences. Some of the mice were found to be positive in seroneutralization assay; in a challenge setting, all but two immunized mice survived, albeit presenting some disease symptoms. 1871 CKPGDPRHC(1) CKPGDPRQC(1) CKPGDPRSC(1) CKAGDPRTC(3) CKVGDPLRC(1) CKVGDPLWC(1) CKLGDPLAC(3) CKLIRPAVC(1) CLICDAKWC(3) CSFAPQFAC(1) CLLVWWAIC(2) K-[LPV]-G-D-P-[RKL] 11 Phage display (common panning) 3 PMID:21764227 Rabbit anti-PrV polyclonal antibody IgG Pseudorabies virus, PrV Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Groups of six C57BL/6 mice were immunized with bacteriophages expressing peptides with this motif sequences. Some of the mice were found to be positive in seroneutralization assay; in a challenge setting, all but two immunized mice survived, albeit presenting some disease symptoms. 1872 LLPGQTTLATGL 1 Phage display (common panning) 3 PMID:21764227 Rabbit anti-PRRSV polyclonal antibody IgG Porcine reproductive and respiratory syndrome virus, PRRSV Not determined. Not determined. Ph.D.-12 phage display library (X12) 1873 SVSVGMKPSPRP(17)[131.1 ± 7.2] FPTRFKKQWSLP(7)[26.7 ± 3.2] WHKSEQGPRLYR(6)[224.2 ± 39.4] FSRPPARHQPHQ(4)[11.5 ± 4.9] FNLPLPSRPLLR(3)[7.5 ± 0.7] HPLCTSHAPVPP(2)[104.0 ± 40.2] NAGDNKFVRVSP(1)[70.5 ± 6.3] 7 Phage display (common panning) 4 PMID:21979823 CD44 antigen protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Binding affinity was determined by ELISA. The Kdapp value (apparent dissociation constant, pM) was obtained from a binding saturation curve obtained from data derived from three independent experiments and shown. Among these, P7 (FNLPLPSRPLLR) was further characterized. Initially, binding affinities of synthesized P7 peptides were assessed by ELISA or fluorescence microplate reader. And the P7 peptide was exhibited a high specificity for CD44, with a low Kd value. 1874 CGLHPAFQC(28) CSMNQPTAC(16) CPHNTTASC(11) CSSHLSSSC(10) CGPIVHSNC(6) CHLPPTIBC(6) CQWSVYNNC(6) CHQVFGAYC(4) CNSLVPPSC(4) CLSHPPTMC(2) CKSETLLSC(2) CHLRAPLLC(2) 12 Phage display (in vivo) 3 PMID:21999821 Rat visceral adipose tissue Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Adipocyte-specific affinity and transdermal activity of the TDA1 (CGLHPAFQC) were validated in vitro and targeting ability of the dermally administered TDA1 to visceral adipose tissue was also confirmed in vivo. TDA1 was effectively translocated into systemic circulation after dermal administration and selectively targeted visceral adipose tissue without any preference to other organs tested. Fluorescent microscopic analysis revealed that the TDA1 could be specifically localized in the hair follicles of the skin, as well as in the visceral adipose tissue. Thus, the authors inferred that dermally administered TDA1 would first access systemic circulation via hair follicles as its transdermal route and then could target visceral fat effectively. The overall results suggest that the TDA1 peptide could be potentially applied as a homing moiety for delivery of anti-obesity therapeutics to visceral fat through the convenient transdermal pathway. 1875 CTQRPDKSC(0.25) 1 Phage display (subtractive panning) 5 PMID:22002548 Hsp70-peptide complexes, Hsp70-PCs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Hsp70-PCs were obtained from the human breast carcinoma cell line MDA-MB-231 using ADP-agarose affinity chromatography. Phages interacting with the HSP70 moiety of the HSP70-PC were removed using a subtractive biopanning step with recombinant HSP70. 1876 CISTHGPLC(1.0) 1 Phage display (subtractive panning) 10 PMID:22002548 Hsp70-peptide complexes, Hsp70-PCs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Hsp70-PCs were obtained from the human breast carcinoma cell line MDA-MB-231 using ADP-agarose affinity chromatography. Phages interacting with the HSP70 moiety of the HSP70-PC were removed using a subtractive biopanning step with recombinant HSP70. One of the peptides (ISTHGPL), termed IST, enriched in the biopanning process, was used in a \'pull-down\' assay to identify the original protein from which the HSP70-associated peptides may have been derived. The eukaryotic translation initiation factor 3 (eIF-3), a member of the elongation factor EF1α family, and the HSP GRP78, were pulled down by theIST peptide. All of these proteins are known to be up-regulated in cancer cells. Immunohistochemical staining of tumour tissue microarrays showed that the peptide co-localised with HSP70 in breast tumour tissue. 1877 STLNVLQ(0.50) TQKWPPH(0.17) HPLIHPD(0.17) APNETRS(0.17) 1 Phage display (subtractive panning) 10 PMID:22002548 Hsp70-peptide complexes, Hsp70-PCs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Hsp70-PCs were obtained from the human breast carcinoma cell line MCF-7 using ADP-agarose affinity chromatography. Phages interacting with the HSP70 moiety of the HSP70-PC were removed using a subtractive biopanning step with recombinant HSP70. 1878 WHWRNPDFWYLK(10)[0.83] SNNLYPQRAVST(1)[0.07] LETEWDSLWYAP(1)[0.11] 3 Phage display (common panning) 5 PMID:22003385 Alanine aminotransferase 1, ALT1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA,Quartz crystal microbalance (QCM) In phage ELISA, ALT binding of the selected phage clones and M13 control phage incubated with varying concentrations of phage particles ranging from e5 pfu to e11 pfu. All measurements were performed in triplicate. The absorbance at 490 nm was reproduced and shown as means of three measurements when the concentration of phage particles is e11 pfu. The absorbance of M13 control phage is 0.07. Besides the apparent dissociation constant (Kd,app, nM) for the ALT5-8 clone with the sequence of WHWRNPDFWYLK is determined by further ELISA experiments. Further, the inhibition constant for the ALT5-8 peptide was found to be KI=71 ± 17 nM. A dissociation constant of Kd= 2.01 ± 0.06 mg/mL or 20.1 ± 0.6 nM was calculated for ALT binding on QCM. Phage particles expressing this peptide exhibited nanomolar affinity for immobilized ALT (Kd,app = 85±20 nM). 1879 LTTKQHNALPRG[0.217] HENPKWHTTPVL[0.243] HMNPKDLVMNSP[0.230] YVVPQSPVRADS[0.321] EHTMAGWWHPSK[0.217] LSPRPLWDAWEQ[NT] SMPSPSYLFGYL[NT] SWSIVPTSPIIL[NT] MNVTVSGRLSGP[NT] TWDLRIHRSVHG[NT] SLPDLRHWSASK[NT] TLSVNSNYGAFT[NT] HENPKQNPLTAR[NT] HANPKQNNHTTS[NT] NPKHIGAEVPQR[NT] YGLNPKWATLKY[NT] SDRISGTSPTRL[NT] IFASFRTSPDQS[NT] HESPKPTVWQTK[NT] 19 Phage display (common panning) 3 PMID:22015270 Anti-SP1 polyclonal antibody IgGs Peptide SP1 (LDLERDARVRAERNANEMSI) of Paramyosin Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Reactivity of anti-SP2 IgG and anti-SP-1 IgG with their selected mimotopes was measured by ELISA. Optical densities at 405 nm were determined, reproduced from the graph and expressed as mean ± SD. The O.D. obtained using an irrelevant phage was 0.064. NT denotes not tested. Antibodies to α-helical regions SP1 were weak binders and M13 phage-displayed peptides selected by them from two different libraries exhibited no amino-acid similarities with the original protein site. 1880 CSLGSQNGC[0.141] CIGPGDRQC[NT] CLAPGDQGC[NT] CLRNALNEC[NT] CNHLHSQRC[NT] CDTRITKRC[NT] CNSWLSHRC[NT] CQHLSSSSC[NT] CSAQSLTLC[NT] CMDPNLPNC[NT] CIDPNRPNC[NT] CKDPNLKNC[NT] CNPKHPLHC[NT] CWQTPSPQC[NT] 14 Phage display (common panning) 3 PMID:22015270 Anti-SP1 polyclonal antibody IgGs Peptide SP1 (LDLERDARVRAERNANEMSI) of Paramyosin Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Reactivity of anti-SP2 IgG and anti-SP-1 IgG with their selected mimotopes was measured by ELISA. Optical densities at 405 nm were determined, reproduced from the graph and expressed as mean ± SD. The O.D. obtained using an irrelevant phage was 0.064. NT denotes not tested. Antibodies to α-helical regions SP1 were weak binders and M13 phage-displayed peptides selected by them from two different libraries exhibited no amino-acid similarities with the original protein site. 1881 DQLLSGSTPWSF[1.098 ± 0.249] AQTLSGTLASHT[1.006 ± 0.046] TTQVLSGTLPQH[1.131 ± 0.117] LHMPPQMLSGTI[0.589 ± 0.109] SQMQILSGSHLA[1.365 ± 0.333] HDQVLSGIKWPH[1.297 ± 0.081] DHSHSNTSGPWV[NT] APEHSQLRHMDP[NT] TIHQVLSATTSH[NT] SVGQTLSGSMLF[NT] QTQMLSGTLIPQ[NT] GQMLSGSNMQLP[NT] MPQILSASRLYA[NT] VPFLPQILSGSL[NT] SLQQLLSGTWLG[NT] SAVLTDIHPVLPNT[NT] KHQVYLRSDLPFNT[NT] SNWRDPLPQTEGNT[NT] Q-x-LSGST 18 Phage display (common panning) 3 PMID:22015270 Anti-SP2 polyclonal antibody IgGs Peptide SP2 (LDELSGTSSQTHDAIRRKDME) of Paramyosin Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Reactivity of anti-SP2 IgG and anti-SP-1 IgG with their selected mimotopes was measured by ELISA. Optical densities at 405 nm were determined, reproduced from the graph and expressed as mean ± SD. The O.D. obtained using an irrelevant phage was 0.064. NT denotes not tested. Antibodies produced in response to non-helical segment within α-helical structure were better binders and selectors of perfect structural mimics of the protein site. 1882 CQQLSRNHC[0.600 ± 0.015] CFQVLSRSC[0.370 ± 0.102] CQRLSGPYC[0.767 ± 0.008] CQKLSQAAC[0.561 ± 0.041] CTQRTQAIC[NT] CPARDQKLC[NT] CTVSLRHVC[NT] CPIQHLSQC[NT] CKTSLKTLC[NT] CQHLSTPVC[NT] CEQTLSGQC[NT] CQTLSKAHC[NT] CQSLSQKLC[NT] CQSLSRPLC[NT] CQSLSFNLC[NT] CQSLSVPRC[NT] CQRLSNNYC[NT] CQKLSANMC[NT] CQKLSMPVC[NT] CQRLSFPIC[NT] CQRLSLPSC[NT] CQKLSLPTC[NT] CQKLSMPAC[NT] CQRLSLAAC[NT] CQRLSTQLC[NT] CQRLSTAHC[NT] CNQQLSRSC[NT] CTQHLSRSC[NT] CPAKNPSAC[NT] CDNTSPSEC[NT] CKYTPQPSC[NT] CNASPQFAC[NT] Q-x-LSGST 32 Phage display (common panning) 3 PMID:22015270 Anti-SP2 polyclonal antibody IgGs Peptide SP2 (LDELSGTSSQTHDAIRRKDME) of Paramyosin Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Reactivity of anti-SP2 IgG and anti-SP-1 IgG with their selected mimotopes was measured by ELISA. Optical densities at 405 nm were determined, reproduced from the graph and expressed as mean ± SD. The O.D. obtained using an irrelevant phage was 0.064. NT denotes not tested. Antibodies produced in response to non-helical segment within α-helical structure were better binders and selectors of perfect structural mimics of the protein site. 1883 QPFTTSLTPPAR(3)[0.416 ± 0.011] LKGPMTDVRESA(2)[0.225 ± 0.002] APLPYDHHSAGS(1)[0.292 ± 0.014] HHQNPPLFLGTS(1)[0.220 ± 0.004] ASLTDRPTLTPV(1)[0.244 ± 0.010] SLNETQHFAFHH(1)[0.264 ± 0.028] TVYSMNSAAPRP(1)[0.272 ± 0.019] SAHGTSTGVPWP(1)[0.204 ± 0.003] NNWSSPPQMISR(1)[0.357 ± 0.017] NFSQPPSKHTRS(1)[0.293 ± 0.010] ATFSPPQQSLMM(1)[0.349 ± 0.019] NSIQALDSTNGQ(1)[0.225 ± 0.022] HPLFHHQTATAT(1)[0.234 ± 0.010] ISTSPPQGSTSS(1)[0.294 ± 0.013] 14 Phage display (competitive panning) 3 PMID:22033953 Mouse cardiomyocyte Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Reactivity of anti-SP2 IgG and anti-SP-1 IgG with their selected mimotopes was measured by ELISA. Optical densities at 405 nm were determined, reproduced from the graph and expressed as mean ± SD. The O.D. obtained using an irrelevant phage was 0.064. NT denotes not tested. The areas with induced beating cardiomyocytes were selected and trypsinized into single cells for the positive screen; undifferentiated mouse embryonic stem cells (mESC) were used as the negative screen. The ELISA results show the no. 3 sequence peptide (QPFTTSLTPPAR), and other four sequences having a consensus motif [SS(Q)PPQ(S)], no. 9, 11, 14, and 10, have relatively high affinity and specificity to cardiomyocytes. Immunofluorescence confirmed that the selected peptides could bind specifically to the PSC-derived cardiomyocytes. Competition tests with chemically synthesized peptides revealed the binding ability was caused by the peptide itself. Western blot analysis proved the phages were both bound to two 17 kDa cardiomyocyte membrane proteins and the no. 9 sequence showed a 55 kDa protein that was not observed in the no. 3 sequence. These results suggest that the selected peptides specifically target receptors on PSC-derived cardiomyocyte membranes. 1884 CRPPR CRKDKC KSTRKS CARSKNKDC 4 Phage display (in vivo) PMID:22047107 Mouse mesenchymal stem cells Not determined. Not determined. Not determined. fUSE5 phage display library Cell coating was optimized and coating persistence and cytotoxicity were evaluated. MSCs were coated with peptides, injected into mice with MI, and MSCs in the heart quantified. Greater numbers of MSCs were found in heart of animals treated with the peptide-coated MSCs compared to uncoated controls. MSC numbers had positive correlation with MI severity in peptide-coated cells but a negative correlation in MSCs alone. A transient cell coating ("painting") method has been developed that labels cells efficiently, non-toxically and increases cell localization in MI heart. 1885 HFFKWHTRTNDQ HMFKWHTRTNDQ HYFKWHTRTNDQ TRVTND 3 Phage display (common panning) 3 PMID:22048717 Anti-CYR61 monoclonal antibody 096B7 Protein CYR61 Not determined. Not determined. Ph.D.-12 phage display library (X12) 1886 CLANFTPHC CLFNFDPTC CLKNWNPHC CLLNCNPQC CLLNFSPYC CLLNWDPQC CLLNWNPHC CLLNWTPHC CLMNWTPHC CLPNWNPHC CLPNWTPQC L-[PL]-N-[WFC]-[NTL]-P-[HQT] 11 Phage display (common panning) 3 PMID:22048717 Anti-CYR61 monoclonal antibody 093G9 Protein CYR61 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 1887 QQPPMHLMSYAG 1 Phage display (subtractive panning) 3 PMID:22071019 Human renal carcinoma cell line A498 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A normal renal cell line HK-2 were used to carry out subtractive screening in vitro. Through a cell-based ELISA, immunocytochemical staining, immunohistochemical staining, and immunofluorescence, the Phage ZT-2 and synthetic peptide ZT-2 were shown to specifically bind to the tumor cell surfaces of A498 and incision specimens, but not to normal renal tissue samples. 1888 ASLRTLTSLLPA NFMESLPRLGMH AHLELRSNNMYF 3 Phage display (common panning) 3 PMID:22065589 Caspase recruitment domain (CARD) of APAF-1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1889 WPTPPYA 1 Phage display (common panning) 3 PMID:22065589 Caspase recruitment domain (CARD) of APAF-1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 1890 CSWFEASYC CLPTLHLLC 2 Phage display (common panning) 3 PMID:22065589 Caspase recruitment domain (CARD) of APAF-1 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 1891 RGWCRPLPKGEG(14) 1 Phage display (subtractive panning) 3 PMID:22086793 Sera from early-stage primary HCC patients Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Microtiter wells were respectively coated overnight with 100ml of purified IgG (100μμg/ml) from each of the control groups. The plates were blocked with 3% nonfat milk for 2 hr, and then washed five times with 0.05% Tween-20 in Tris-buffered saline (PBST). One hundred ?`l of diluted random 12-peptide phage display library with a titer of 1.5e11 pfu/ml was added to coated plates. After sequential incubation for 1 hr at room temperature with each of the control IgG-coated plates, the unbound phages were collected and added 100μl/well into the early HCC-IgG-coated plate. Another two rounds of affinity selection were carried out in the same way, except that 1:200 and 1:400 sera dilution was added to 100μl of diluted phages from the last round. In the screening phase, 19 out of 20 randomly selected phage clones exhibited specific reaction with purified sera IgG from early primary HCC patients, among them 14 coming from the same phage clone with inserted peptide sequence RGWCRPLPKGEG (named HC1). The HC1 mimic peptide showed high diagnostic validity for early primary HCC, and thereby could be a candidate serum biomarker for early primary HCC. 1892 KMSNSIY KLWVIPQ KLYTPVD 3 Phage display (subtractive panning) 4 PMID:22092230 Human chronic myeloid leukemia (CML) cell lines KT-1/A3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) IFN-a-sensitive KT-1/A3 cells were used as the target, and IFN-a-resistant subline KT-1/A3R was used as absorber for phage display biopanning. Totally, 210 phage clones were picked for screening whole-cell ELISA, among which six phages were differently reactive against KT-1/A3 and KT-1/A3R cells. Multiple clones showed high binding efficiency to KT-1/A3 cells compared with that of the other leukemia cells. One of the peptides, KLWVIPQ, has a partial amino acid sequence homology with the C-terminal domain of E3 ubiquitin-protein ligase. 1893 TFLPQPRCSALLRYLSEDGVIVPS 1 Phage display (common panning) 4 PMID:10411639 Urease Not determined. Not determined. Not determined. X10-ALLRY-X10 phage display library ELISA,Competition experiment The absorbance was measured at 450 nm (data not shown). About 60% of the clones selected from the combinatorial library gave a positive ELISA signal. The 24-mer peptide inhibits the activity of H. pylori urease holoenzyme in a competitive manner. The calculated inhibition constant (Ki) was 49 μm. The same 24-amino-acid peptide sequence (TFLPQPRCSALLRYLSEDGVIVPS) was found in all of the 10 phage clones isolated from the combinatorial library. 1894 YDFYWW 1 Phage display (common panning) 4 PMID:10411639 Urease Not determined. Not determined. Not determined. X6 fdMED1 phage display library ELISA,Competition experiment The absorbance was measured at 450 nm (data not shown). About 20% of the clones selected from the unconstrained hexapeptide library gave a positive ELISA signal. The hexapeptide inhibits urease activity in an uncompetitive manner and the calculated inhibition constant (Ki) was 30 μm. The same six-amino-acid peptide sequence (YDFYWW) in the 10 clones were selected from the unconstrained library. 1895 CQSPSLQC CVFQNLDC CKDGGNAC 3 Phage display (common panning) 4 PMID:10411639 Urease Not determined. Not determined. Not determined. CX6C fdMED1 phage display library 1896 CRLKEKHC(10) 1 Phage display (common panning) 3 PMID:22100634 Anti-CD11b monoclonal antibody 44a Integrin alpha-M, integrin αM Not determined. Not determined. CX6C fdMED1 phage display library Independent phage clones (94 bacterial clones) from the third round of selection were grown individually and their phage retested for binding to mAb 44a (IgG 2a) and to murine irrelevant IgG2a. Sixty-two pIII-cys-6aa-cys clones bound, specifically to mAb 44a, but not by no reactivity with isotype control was observed. The DNA inserts of the 10 phage clones giving the strongest ELISA signals were sequenced, and their amino acid sequences were deduced. One peptide sequence C-RLKEKH-C (frequency=10/10) which appears to be responsible for the specific binding to mAb 44a was identified. The selected peptide mimics a discontinuous epitope, a peculiar shape on the CD11b-I-domain surface. Competitive ELISA experiments with different Mac-1 ligands showed that C-RLKEKH-C is able to bind to fibrinogen, iC3b, and C1q. 1897 YTSPHHSTTGHL SMMLPFPHQPNA AVPHRVGGLHSL 3 Phage display (subtractive panning) 4 PMID:22107959 MBP mouse Kelch protein (MBP-Kelch) Nuclear factor erythroid 2-related factor 2 1X2R,2DYH, Not determined. Ph.D.-12 phage display library (X12) In forth round, the bound phage were eluted with ETGE peptide. MBP and MBP-Kelch(G364C) were used to carry out subtractive screening. 1898 GSSTGPQRLHVP 1 Phage display (subtractive panning) 4 PMID:22107959 MBP mouse Kelch protein (MBP-Kelch) Nuclear factor erythroid 2-related factor 2 1X2R,2DYH, Not determined. Ph.D.-12 phage display library (X12) In forth round, the bound phage were eluted with ETGE peptide. MBP and MBP-Kelch(G430C) were used to carry out subtractive screening. 1899 NTMHYTGHTHSP HAKSQLNPPDIK GSSTGPQRLHVP TLDLPGWTIVSH 4 Phage display (subtractive panning) 4 PMID:22107959 MBP mouse Kelch protein (MBP-Kelch) Nuclear factor erythroid 2-related factor 2 1X2R,2DYH, Not determined. Ph.D.-12 phage display library (X12) In forth round, MBP and MBP-Kelch(G364C) were used to carry out subtractive screening. 1900 NTMHYTGHTHSP SPNFSWLPLGTT SILSTMSPHGAT TPKNLPPPGQRA HLPTHVRTIVGL SYINNTPVVDRR 6 Phage display (subtractive panning) 4 PMID:22107959 MBP mouse Kelch protein (MBP-Kelch) Nuclear factor erythroid 2-related factor 2 1X2R,2DYH, Not determined. Ph.D.-12 phage display library (X12) In forth round, MBP and MBP-Kelch(G430C) were used to carry out subtractive screening. 1901 SMMKADFDEEPR(24) SMMKADFEEEPR(8) 2 Phage display (subtractive panning) 3 PMID:22123204 Anti-Bla g 2 monoclonal antibody 7C11 Aspartic protease Bla g 2 2NR6, Not determined. Ph.D.-12 phage display library (X12) Phages were precleared by incubation for 20 min at room temperature with mouse IgG mAb 6G-2 of unrelated specificity bound to protein G beads. Unbound phages were then incubated with 300 ng of purified mAb 7C11. The amino acid sequences of the peptides on selected phages differed at only one position, occupied by 1 of 2 negatively charged residues. The two 12-mer sequences bound to 7C11 with similar avidity and specificity. There was good concordance between the residues in the 3D clusters identified from our phage display/computational method with the co-crystal structural analysis. The Episearch analysis using peptide 1 identified 2 high-scoring patches centered at residue Asp100 (score = 1.0) and Asp87 (score = 0.87), respectively. Analysis with peptide 2 identified 1 high-scoring patch, centered at Pro61 (score = 1.0), and 1 low-scoring patch, centered at Arg83 (score = 0.67). The same 4 patches, centered at Asp100, Pro61, Asp87 and Arg83, were also identified using the sequences of peptides 1 and 2 simultaneously. After antibody binding, amino acids Lys60, Lys65, Ile67, Asp68 and Arg83 lost more than 50 Å(2) of their surface area, while Try66 and Lys132 lost 100 Å(2) or more of their surface area. 1902 SYPIPDT(14) HTSDQTN(9) AKLPIIP(5) AYHPPAM(4) EGMHYHT(2) EPAHMSL(1) WTPYSLV(1) SNYLMYI(1) HATSYFV(1) 9 Phage display (subtractive panning) 4 PMID:22136656 Epidermal growth factor receptor Pro-epidermal growth factor, EGF 1IVO,1NQL,3NJP, Not determined. Ph.D.-7 phage display library (X7) A-431 cells expressing epidermal growth factor receptor were used as the matrix in a cell-based subtractive biopanning approach using a 7-mer peptide displaying phage library. Human fetal foreskin fibroblast cells were used to carry out subtractive screening. Competitive binding of the selected peptides (SYPIPDT and HTSDQTN) was performed to determine the affinities of the peptides displayed by the phage particles toward EGFR. Two novel peptide ligands were identified and tested for their affinities and functional effects on epidermal growth factor receptor. The identified peptides were able to inhibit the epidermal growth factor-induced phosphorylation of epidermal growth factor receptor in a concentration-dependent manner. The results of affinity binding experiments showed that the natural ligand, that is epidermal growth factor, was able to inhibit competitively the binding of peptide-bearing phage to epidermal growth factor receptor expressing A-431 cells. Molecular modeling studies were used to calculate the free energies for the binding of peptides to the receptor-binding site as well as proposing the interaction modes for this binding. 1903 HPLSKHPYWSQP 1 Phage display (common panning) 3 PMID:22119929 Clusterin Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Competition experiment,NMR spectroscopy The phage-displayed P3378 (HPLSKHPYWSQP) and synthetic P3378 (HPLSKHPYWSQP) bind in a competitive manner. Besides, the results of DRP-NMR strongly suggest that the P3378 peptide and not the P3378R peptide (PYLHQSPHWKPS), interacts with rhsCLU in a sequence-specific manner. HEK293-E6 cells expressing recombinant human secreted (rhs)CLU were used as the target in a cell-based biopanning approach. A third panning round returned phage clones of which essentially 100% displayed the same peptide. Sequencing of the peptide revealed the unique amino acid sequence HPLSKHPYWSQP, which was designated peptide P3378. Differential resonance perturbation nuclear magnetic resonance using P3378 and a scrambled control peptide (designated P3378R) confirmed the P3378-sCLU interaction and demonstrated that it was sequence specific. P3378 and P3378R peptides were conjugated to an Alexa680 near infrared fluorophore (NIRF) and assessed for their tumor homing abilities in in vivo time-domain fluorescence optical imaging experiments using living 4T1 tumor bearing BALB/c mice. When injected in separate animals, both peptides accumulated at the tumor site, however the NIRF-labeled P3378 peptide was retained for a significant longer period of time than the P3378R peptide. Similar observations were made after simultaneously injecting the same tumor-bearing animal with a peptide mixture of P3378 DyLight (DL)680 and the P3378R-DL800. Coinjection of P3378-DL680 with excess unlabeled P3378 blocked tumor accumulation of fluorescent signal while excess P3378R control peptide did not confirming the sequence specificity of the tumor accumulation. Finally, ex vivo fluorescence microscopy of these tumors confirmed the presence of P3378-DL680 in the tumor and its colocalization with CLU. These results confirm the tumor targeting specificity of the P3378 CLU-binding peptide and suggest its usefulness for the in vivo monitoring of solid tumors secreting detectable levels of CLU. 1904 PSTRKK(1) TGGWRF(2) QWPMLV(1) WSGQDI(1) YRRGHL(1) RYRTSL(2) SWGRRG(1) VRKVAA(2) SRSCTA(1) LEALVV(1) SACLTG(1) CLKTEG(1) FRRPGG(1) RGWGTV(1) LRYVTY(1) GGRVVT(1) RVYLLS(1) RAQMGE(1) PVQKKH(1) VGRHLA(1) KGWEGC(1) 21 Phage display (common panning) 5 PMID:22132161 Serine protease 28 Not determined. Not determined. Not determined. X6 T7 phage display library Phage display peptide mimetics revealed an expanded but mixed substrate specificity of ISP1, including chymotryptic and elastase activity. Based upon targets observed using phage display, we hypothesised that ISP1 might signal to cells by cleaving and activating proteinase-activated receptors (PARs) and therefore assessed PARs 1, 2 and 4 as potential ISP1 targets. 1905 FHKHKSPALSPV(19) FHKPFFPKGSAR(13) 2 Phage display (common panning) 3 PMID:22138765 Tenascin, TN Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Most solid tumors express the large isoform of TNC which contains EGF domain, fibrinogen glob (fbg) and fibronectin III (FN III) domains with the inclusion of alternatively spliced exons in the middle. Due to the complexity and sizes of the tumor specific TNC protein, the authors utilized two different versions of TNC proteins and performed two independent selections. One is the tumor-specific large isoform of TNC (Full-length TNC) expressed in eukaryotic cells and the other is the alternatively spliced domain (A1-D) of TNC expressed in bacteria. Of the 35 clones derived from two independent selections, 19 had the same sequences (designated peptide #1), and 13 others were also identical (designated peptide #2), while the remainder had similar sequences. The selected peptide #1 specifically recognized tenascin C protein in xenograft mouse tissue. The authors also observed exclusive staining of tenascin C by the selected peptide in tumor patient tissues. Moreover, the peptide reduced tenascin C-induced cell rounding and migration. 1906 HVHPPLRPHSDK(1) YPTHHAHTTPVR(10) HATNHLPTPHNR(1) 3 Phage display (subtractive panning) 5 PMID:22163050 Bovine rotavirus Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the fourth round of panning, the coated viruses were replaced by the supernatant of MA104 culture. The phages were incubated with the supernatant at room temperature for 30 min prior to the fifth round of panning. Using the specific peptide-expressing phages, the authors developed a phage-based ELISA to differentiate BRV from other viruses. Compared with quantitative real-time PCR (qPCR), the phage-mediated ELISA was more suitable for the capture of BRV and the detection limitation of this approach was 0.1 μg/ml of samples. The high sensitivity, specificity and low cross-reactivity for the phage-based ELISA were confirmed in receiver operating characteristics (ROC) analysis. 1907 WHWRLPS(10) LHHKTHH(2) 2 Phage display (common panning) 3 PMID:22171803 Anti-influenza A H2N2 monoclonal antibody C179 Anfluenza A virus strain H2N2 Not determined. Not determined. Ph.D.-7 phage display library (X7) 1908 WHTHKWSLSAKA(4) HHWKFFFSHPGA(2) HHWKFFFSHPGE(2) LPFHGHKKPVLS(1) WPWWPGHTHRTI(1) HPMKQYRWRPSI(1) SPNYWFNKIHQH(1) 7 Phage display (common panning) 3 PMID:22171803 Anti-influenza A H2N2 monoclonal antibody C179 Anfluenza A virus strain H2N2 Not determined. Not determined. Ph.D.-12 phage display library (X12) 1909 CNLSSSWIC(9) CNSGMFVRC(3) 2 Phage display (common panning) 3 PMID:22171803 Anti-influenza A H2N2 monoclonal antibody C179 Anfluenza A virus strain H2N2 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 1910 QWTWTQY(4) DTLPLFI(1) MSLQQEH(1) ANTTPRH(1) MDAHHAL(1) ITAPHPH(1) QRNQTQD(1) QWNRTQY(1) NTAPHPH(1) 9 Phage display (common panning) 3 PMID:22171803 Anti-influenza A H1N1 monoclonal antibody IV.C102 Anfluenza A virus strain H1N1 Not determined. Not determined. Ph.D.-7 phage display library (X7) 1911 DCWQMDRKTCPL(6) NTPAWLNHTTVI(3) LPAFFVTNQTQD(1) TVHWWZTHGPLS(1) SAIPTTWNPLAV(1) 5 Phage display (common panning) 3 PMID:22171803 Anti-influenza A H1N1 monoclonal antibody IV.C102 Anfluenza A virus strain H1N1 Not determined. Not determined. Ph.D.-12 phage display library (X12) 1912 CPLHARLPC(10) CSLASLPAC(2) 2 Phage display (common panning) 3 PMID:22171803 Anti-influenza A H1N1 monoclonal antibody IV.C102 Anfluenza A virus strain H1N1 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 1913 WPWHNHR(6) ASINSSL(2) QSERAIQ(1) TSLPTIV(1) AFSYHIH(1) NMWQALN(1) 6 Phage display (subtractive panning) 3 PMID:22171803 Anti-influenza A H3N2 polyclonal antibody IgG Influenza A virus (strain A/Texas/1/1977 H3N2) Not determined. Not determined. Ph.D.-7 phage display library (X7) Goat IgG were used in a subtractive biopanning approach. 1914 VWSTPPHADGPA(4) HAPWRHHQASPK(3) FPAHPAWTIGSM(1) YTPLSSASPWGP(1) GMSLLHGQRPHT(1) VSRHQSWHPHDL(1) EREAHQLHSHHK(1) 7 Phage display (subtractive panning) 3 PMID:22171803 Anti-influenza A H3N2 polyclonal antibody IgG Influenza A virus (strain A/Texas/1/1977 H3N2) Not determined. Not determined. Ph.D.-12 phage display library (X12) Goat IgG were used in a subtractive biopanning approach. 1915 CLGALSHTC(8) CSPVLPFLC(1) CTHEPSGRC(1) CSAPRQADC(1) CSLPLTGQC(1) 5 Phage display (subtractive panning) 3 PMID:22171803 Anti-influenza A H3N2 polyclonal antibody IgG Influenza A virus (strain A/Texas/1/1977 H3N2) Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Goat IgG were used in a subtractive biopanning approach. 1916 ETKAWWL(7) LASKPMP(4) QAHTIST(1) 3 Phage display (subtractive panning) 3 PMID:22171803 SIV sera from patients Swine-origin influenza virus A/2009 Not determined. Not determined. Ph.D.-7 phage display library (X7) Human IgG were used in a subtractive biopanning approach. 1917 QAHNWYNHKPLP(6) VHNNAARTGSPP(2) VHNHANDPGSPP(1) QELYPYSPHIHV(1) FSHELSWKPRKA(1) AHTHSKERVQTI(1) 6 Phage display (subtractive panning) 3 PMID:22171803 SIV sera from patients Swine-origin influenza virus A/2009 Not determined. Not determined. Ph.D.-12 phage display library (X12) Human IgG were used in a subtractive biopanning approach. 1918 CPLHARLPC(6) CRHLPLTPC(1) CSLPLTGQC(1) CPSYPLSFC(1) CRDISPLAC(1) CYGWPIYSC(1) CNLSSSWTC(1) 7 Phage display (subtractive panning) 3 PMID:22171803 SIV sera from patients Swine-origin influenza virus A/2009 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Human IgG were used in a subtractive biopanning approach. 1919 CLRSGRFC(10) 1 Phage display (competitive panning) 3 PMID:22189418 C-X-C chemokine receptor type 1, CXCR-1 Interleukin-8, IL-8 1ILP,1ILQ, Not determined. CX6C fdMED1 phage display library In fact, hCXCR1-transfected murine pre-B cells were used to panning the phage library. In each round, bound phage were eluted with hCXCL8. Sequence analysis revealed homology between the linear hexapeptides and the N-terminal domain (1-SAKELR-6), whereas the constrained peptides are composed of non-contiguous amino acids mimicking spatial structure on the surface of folded C-terminal portion of hCXCL8 (50-CLDPKENWVQRVVEKFLKRAENS-72). The synthetic linear and structurally constrained peptides competed for 125I-hCXCL8 binding to hCXCR1 and hCXCR2 (IC50 comprised between 10 and 100 lM). Furthermore, they inhibited the intracellular calcium flux and the migration of hCXCR1/hCXCR2 transfectants; and desensitized hCXCR1 and hCXCR2 receptors on neutrophils, reducing their chemotactic responses induced by ELR-CXC chemokines (hCXCL8, hCXCL1, hCXCL2, hCXCL3, and hCXCL5). 1920 MSRAKE(10) 1 Phage display (competitive panning) 3 PMID:22189418 C-X-C chemokine receptor type 1, CXCR-1 Interleukin-8, IL-8 1ILP,1ILQ, Not determined. X6 fdMED1 phage display library In fact, hCXCR1-transfected murine pre-B cells were used to panning the phage library. In each round, bound phage were eluted with hCXCL8. Sequence analysis revealed homology between the linear hexapeptides and the N-terminal domain (1-SAKELR-6), whereas the constrained peptides are composed of non-contiguous amino acids mimicking spatial structure on the surface of folded C-terminal portion of hCXCL8 (50-CLDPKENWVQRVVEKFLKRAENS-72). The synthetic linear and structurally constrained peptides competed for 125I-hCXCL8 binding to hCXCR1 and hCXCR2 (IC50 comprised between 10 and 100 lM). Furthermore, they inhibited the intracellular calcium flux and the migration of hCXCR1/hCXCR2 transfectants; and desensitized hCXCR1 and hCXCR2 receptors on neutrophils, reducing their chemotactic responses induced by ELR-CXC chemokines (hCXCL8, hCXCL1, hCXCL2, hCXCL3, and hCXCL5). 1921 CLPWKENC(10) 1 Phage display (competitive panning) 3 PMID:22189418 C-X-C chemokine receptor type 2, CXCR-2 Interleukin-8, IL-8 Not determined. Not determined. CX6C fdMED1 phage display library In fact, hCXCR2-transfected murine pre-B cells were used to panning the phage library. In each round, bound phage were eluted with hCXCL8. Sequence analysis revealed homology between the linear hexapeptides and the N-terminal domain (1-SAKELR-6), whereas the constrained peptides are composed of non-contiguous amino acids mimicking spatial structure on the surface of folded C-terminal portion of hCXCL8 (50-CLDPKENWVQRVVEKFLKRAENS-72). The synthetic linear and structurally constrained peptides competed for 125I-hCXCL8 binding to hCXCR1 and hCXCR2 (IC50 comprised between 10 and 100 lM). Furthermore, they inhibited the intracellular calcium flux and the migration of hCXCR1/hCXCR2 transfectants; and desensitized hCXCR1 and hCXCR2 receptors on neutrophils, reducing their chemotactic responses induced by ELR-CXC chemokines (hCXCL8, hCXCL1, hCXCL2, hCXCL3, and hCXCL5). 1922 CAKELR(10) 1 Phage display (competitive panning) 3 PMID:22189418 C-X-C chemokine receptor type 2, CXCR-2 Interleukin-8, IL-8 Not determined. Not determined. X6 fdMED1 phage display library In fact, hCXCR2-transfected murine pre-B cells were used to panning the phage library. In each round, bound phage were eluted with hCXCL8. Sequence analysis revealed homology between the linear hexapeptides and the N-terminal domain (1-SAKELR-6), whereas the constrained peptides are composed of non-contiguous amino acids mimicking spatial structure on the surface of folded C-terminal portion of hCXCL8 (50-CLDPKENWVQRVVEKFLKRAENS-72). The synthetic linear and structurally constrained peptides competed for 125I-hCXCL8 binding to hCXCR1 and hCXCR2 (IC50 comprised between 10 and 100 lM). Furthermore, they inhibited the intracellular calcium flux and the migration of hCXCR1/hCXCR2 transfectants; and desensitized hCXCR1 and hCXCR2 receptors on neutrophils, reducing their chemotactic responses induced by ELR-CXC chemokines (hCXCL8, hCXCL1, hCXCL2, hCXCL3, and hCXCL5). 1923 MLRQTR(2) HASILP(5) KKEPWI(3) 3 Phage display (common panning) 3 PMID:22189418 Interleukin-8, IL-8 Not determined. Not determined. Not determined. X6 fdMED1 phage display library These peptides similarly displaced the binding of 125I-hCXCL8 to hCXCR1 in a dose-dependent manner, inhibited hCXCL8 induced increases in the intracellular calcium, and migration of hCXCR1- and hCXCR2-transfected cells. 1924 LSTHTTESRSMV LEPRWGFGWWLK 2 Phage display (common panning) 3 PMID:22227343 Prestin Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Prestin was adsorbed to polypropylene Petri dishes, these were used to perform the first two of three rounds of biopanning. The 3rd round of biopanning was performed using Cos-7 cells transiently transfected with a cCFP-prestin plasmid, to eliminate false-positives affinitive to intracellular loops and trans-membrane helices of the prestin that may have been obtained using soluble prestin. The binding properties of LSTHTTESRSMV and LEPRWGFGWWLK shown by flow cytometry demonstrated selectivity to prestin-expressing Chinese hamster ovary cells. PEG6K-b-PCL19K polymersomes covalently labelled with these peptides demonstrated effective targeting to outer hair cells in a rat cochlear explant study. 1925 IMVTESSDYSSY 1 Phage display (common panning) 4 PMID:22233341 Graphite flakes Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) As a novel approach, the authors demonstrate here that short dodecapeptides selected by phage display are capable of self-assembly on graphite and form long-range-ordered biomolecular nanostructures. Using atomic force microscopy and contact angle studies, they identify three amino acid domains along the primary sequence that steer peptide ordering and lead to nanostructures with uniformly displayed residues. The peptides are further engineered via simple mutations to control fundamental interfacial processes, including initial binding, surface aggregation and growth kinetics, and intermolecular interactions. Tailoring short peptides via their primary sequence offers versatile control over molecular self-assembly, resulting in well-defined surface properties essential in building engineered, chemically rich, bio-solid interfaces. 1926 TGLESGHGPGDS 1 Phage display (subtractive panning) 3 PMID:22236882 Purified immunoglobulin G (IgG) from sera of knee OA patients Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) To increase the screening specificity, microtiter wells were coated with purified sera IgG from healthy controls to absorb non-specific phages from the phage random peptide library, and the unbounded phages were then incubated with sera IgG from knee OA patients to screen for phages containing knee OA-specific peptides. The phage clone with inserted peptide TGLESGHGPGDS (named KOA1) showed 90% positive reaction rate with the knee OA patients, significantly higher than that with the knee RA patients (27.8%), the nonerosive hand OA patients (34.3%), the erosive hand OA patients (31.3%) and the healthy controls (12.0%), but not the hip OA patients (82.5%). The novel knee OA mimic peptide KOA1 could be a potential serum biomarker for knee OA. 1927 NHWSDKRAQITI(6) 1 Phage display (common panning) 5 PMID:22239472 Gadolinium oxide (GdO) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) These peptide adhesion domains are exploited to effectively decorate GdO particles with fluorescently labeled poly(ethylene oxide) (PEO), proving to result in a stable surface modification as shown by significant reduction of protein adsorption by 80%, compared to nonfunctionalized particles. 1928 CHMRNTIHC(1) CHSSLLNPC(1) CNSKLHAFC(1) CYNPRLHIC(1) CMEPKSRYC(1) CSNRMATTC(1) 3 Phage display (subtractive panning) 3 PMID:22249604 Anti-gp75 monoclonal antibody 5E7C P. brasiliensis 75-kDa protein Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Binding assay The interaction between the selected phage clones and the 5E7C mAb was evaluated by in vitro binding assays. Binding assay showed that phage displaying P13 (CHSSLLNPC) is a putative mimotope to anti gp75 mAb. Library was cleaned with mAb 17C (anti-gp43) that belongs to the same isotype of the mAb 5E7C (IgG2a) to eliminate non-specific phage clones to the target molecule (mAb 5E7C). Aiming to improve the antigenic potential of the peptide selected sequence, five amino acids similar to the phosphatase sequence showing greater homology were added to the N-terminal and C-terminal, flanking the initial sequence of nine amino acids. That approach led to a synthetic peptide (NANEAHSSLLNNALSI) named P2, which was synthesized and tested against PCM patients' sera to check whether it was recognized. There was significant recognition of P2 by sera of untreated PCM patients when compared with normal human sera. Sera from treated PCM group, patients with other mycosis or co-infected with HIV had much lower recognition of P2 than untreated patient group. The test showed a sensitivity of 100 and 94.59% of specificity in relation to human sera control. These data indicate a potential use of P2 as diagnostic tool in PCM. Its application for serological diagnosis of PCM may contribute to the development and standardization of simpler, faster and highly reproducible immunodiagnostic tests at low cost. 1929 DSGLCMPRLRGCDPR 1 Phage display (common panning) PMID:22257077 Low-density lipoprotein receptor Not determined. Not determined. Not determined. Not determined. From an initial phage display biopanning, a series of peptide ligands for the LDLR was optimized leading to size reduction and improved receptor binding affinity with the identification of peptide 22 and its analogues. Further real-time biphoton microscopy experiments on living mice demonstrated the ability of peptide 22 to efficiently and quickly cross CNS physiological barriers. This validation of peptide 22 led us to explore its binding on the extracellular LDLR domain from an NMR-oriented structural study and docking experiments. 1930 WFHCPYDLCHIL QWECPYGLCWIQ 2 Phage display (subtractive panning) 4 PMID:22263840 Soluble fibrin fragment DD(E) Not determined. Not determined. Not determined. TN6 phage display library Before each round, libraries were depleted of fibrinogen binders by incubating the phage with biotinylated fibrinogen bound to bead immobilized streptavidin. 1931 RSCNYYGTCLH HDCQYYGTCLH FACHYYGTCLH RPCDYYGTCFD 4 Phage display (subtractive panning) 4 PMID:22263840 Soluble fibrin fragment DD(E) Not determined. Not determined. Not determined. TN7 phage display library Before each round, libraries were depleted of fibrinogen binders by incubating the phage with biotinylated fibrinogen bound to bead immobilized streptavidin. 1932 LPCDYYGTCLD LPCSYYGTCLH LSCDYYGTCLR LACHYYGTCLH RPCNYYGTCLH DPCSYYGTCLH FACHYYGTCLH 7 Phage display (subtractive panning) 4 PMID:22263840 Fibrin Not determined. Not determined. Not determined. TN7 phage display library Before each round, libraries were depleted of fibrinogen binders by incubating the phage with biotinylated fibrinogen bound to bead immobilized streptavidin. 1933 RFCNPFAWLCFD HFCSPFHLHCFR HFCNPFFFPCLH HFCSPFLLDCPH RFCNPFLLDCAH HFCFWPIGNCPH 6 Phage display (subtractive panning) 4 PMID:22263840 Fibrin Not determined. Not determined. Not determined. TN8 phage display library Before each round, libraries were depleted of fibrinogen binders by incubating the phage with biotinylated fibrinogen bound to bead immobilized streptavidin. 1934 DCDDHWPRWCH FCRHFNFSFCF 2 Phage display (subtractive panning) 4 PMID:22263840 Fibrin Not determined. Not determined. Not determined. TN9 phage display library Before each round, libraries were depleted of fibrinogen binders by incubating the phage with biotinylated fibrinogen bound to bead immobilized streptavidin. 1935 NHGCYNSYGVPYCDYS 1 Phage display (subtractive panning) 4 PMID:22263840 Soluble fibrin fragment DD(E) Not determined. Not determined. Not determined. TN10 phage display library Before each round, libraries were depleted of fibrinogen binders by incubating the phage with biotinylated fibrinogen bound to bead immobilized streptavidin. 1936 GPRPLSNSVSAI GPRPNSPAPAGS 2 Phage display (subtractive panning) 4 PMID:22263840 Soluble fibrin fragment DD(E) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Before each round, libraries were depleted of fibrinogen binders by incubating the phage with biotinylated fibrinogen bound to bead immobilized streptavidin. 1937 GPRPTSS GPRPTYM GPRPPMV GPRPPTD GPRPPYA GPRPPTI GPRPPDW 7 Phage display (subtractive panning) 4 PMID:22263840 Soluble fibrin fragment DD(E) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Before each round, libraries were depleted of fibrinogen binders by incubating the phage with biotinylated fibrinogen bound to bead immobilized streptavidin. 1938 CHPMAPKYC CHPMAPRYC CHPQAGASC 3 Phage display (subtractive panning) 4 PMID:22263840 Soluble fibrin fragment DD(E) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Before each round, libraries were depleted of fibrinogen binders by incubating the phage with biotinylated fibrinogen bound to bead immobilized streptavidin. 1939 HWGNHSKSHPQR(20) KSLSRHDHIHHH(5) 2 Phage display (common panning) 4 PMID:22265946 Anti-RPV-H monoclonal antibody C1 Hemagglutinin glycoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) Four independent peptide library screenings yielded 80 fourth-round phage clones that were selected by the C1 mAb. In twenty phage clones the amino acid sequence HWGNHSKSHPQR (Domain 1 (D1)) was obtained, whilst five phage clones had the amino acid sequence KSLSRHDHIHHH (D2). The alignment analysis with the RPV RBOK H protein revealed that residues within D1 aligned to amino acids 309-320 in H, whereas residues within D2 aligned to amino acids 529-540. 1940 YQVTQSKVMSHR[0.479 ± 0.079] YQVTPIKLISHI[0.526 ± 0.046] NHVTAIKLITHI[0.521 ± 0.048] YQVTQSKEISHR[0.706 ± 0.038] YHVTPIKLISHI[0.613 ± 0.056] 4 Phage display (common panning) 3 PMID:22314514 Phosphocarrier protein HPr Phosphoenolpyruvate-protein phosphotransferase 2XDF,3EZA,3EZB,3EZE, Not determined. Ph.D.-12 phage display library (X12) ELISA The binding of selected clones to phosphocarrier protein HPr was confirmed by ELISA. The experiments were performed triplicate. Absorbance at 410 nm was determined, reproduced from the graph and expressed as the mean ± SD. The original library was served as the control (with A410 of 0.214 ± 0.040). The single-stranded DNAs of the 20 selected phages were isolated, sequenced, and five corresponding peptides were synthesized. Only one of the five peptides, AP1 (YQVTQSKVMSHR) was found to inhibit the growth of Escherichia coli cells efficiently (IC50~50 μM). Molecular modeling reveals that AP1 may block the EI-HPr interaction and phosphotransfer. Interestingly, AP1 was also found to induce cell aggregation in a concentration-dependent manner. 1941 EPLQLKM(3) STAPRPY(3) SPPQSRA(3) TTPTKSA(2) SPLSEHS(2) IPTLPSS(1) HWGMWSY(1) QPLQLQV(1) SPLQMMQ(1) LTIPTTA(1) VVGTANT(1) MYASSSK(1) 12 Phage display (subtractive panning) 2 PMID:22322196 Human bone marrow-derived mesenchymal stem cell, hBMMSCs Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the negative selection procedure, the library was incubated with the synovial cells for 1 h to exclude the synovial cell affinity phage clones. The authors identified an MSC-homing peptide EPLQLKM (E7) through phage display and constructed an MSC-homing device by conjugating E7 with PCL electrospun meshes. The E7-conjugated PCL electrospun meshes have shown an apparent MSC-homing property in vivo. In addition, the E7-conjugated PCL electrospun meshes have shown specificity toward MSCs compared with the RGD-conjugated PCL electrospun meshes. The current study offers a promising way to improve MSC-based TE repair. 1942 EPLQLKM(7) IPTLPSS(6) MYASSSK(2) QPWPTSI(2) SPPQSRA(1) HWGMWSY(1) 6 Phage display (subtractive panning) 3 PMID:22322196 Human bone marrow-derived mesenchymal stem cell, hBMMSCs Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the negative selection procedure, the library was incubated with the synovial cells for 1 h to exclude the synovial cell affinity phage clones. The authors identified an MSC-homing peptide EPLQLKM (E7) through phage display and constructed an MSC-homing device by conjugating E7 with PCL electrospun meshes. The E7-conjugated PCL electrospun meshes have shown an apparent MSC-homing property in vivo. In addition, the E7-conjugated PCL electrospun meshes have shown specificity toward MSCs compared with the RGD-conjugated PCL electrospun meshes. The current study offers a promising way to improve MSC-based TE repair. 1943 EPLQLKM(10) IPTLPSS(4) HWGMWSY(3) LTIPTTA(2) MYASSSK(1) 5 Phage display (subtractive panning) 4 PMID:22322196 Human bone marrow-derived mesenchymal stem cell, hBMMSCs Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the negative selection procedure, the library was incubated with the synovial cells for 1 h to exclude the synovial cell affinity phage clones. The authors identified an MSC-homing peptide EPLQLKM (E7) through phage display and constructed an MSC-homing device by conjugating E7 with PCL electrospun meshes. The E7-conjugated PCL electrospun meshes have shown an apparent MSC-homing property in vivo. In addition, the E7-conjugated PCL electrospun meshes have shown specificity toward MSCs compared with the RGD-conjugated PCL electrospun meshes. The current study offers a promising way to improve MSC-based TE repair. 1944 FHWTWQFPYTST(3) FHWNYYLYSQVS(3) WHWQWWLQTDAT(2) FHWWPPSLANQP(1) WHWNAWNWSSQQ(1) 5 Phage display (common panning) 3 PMID:22322353 Whole Mycobacterium bovis cells, WCA Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1945 SEFPRSWDMETN(7) FEFPRSWDMETN(1) NFRVSIDVVKSR(1) 3 Phage display (common panning) 3 PMID:22322353 Ethanol-extracted Mycobacterium bovis cell surface antigens, EEA Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1946 GNLLHNHETYRH(2) HSLRWDWTARNS(1) HSLRDDIRTMTA(1) HKWGGNTLMAFR(1) HKPPTHIYLSWR(1) KVWPNMFANENI(1) KLWSIPKDLGPP(1) GYVHRELAWNMN(1) NMELHPHSLPRP(1) 9 Phage display (common panning) 3 PMID:22322353 Cell surface lipoprotein MPB83 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1947 QEINSSY[0.65 ± 0.02] SHPRLSA[0.72 ± 0.05] SMPNPMV[0.69] GLQQVLL[0.79 ± 0.04] HELSVLL[0.63 ± 0.04] YAPQRLP[0.58] TPRTLPT[0.51 ± 0.03] APVHSSI[0.45 ± 0.02] APPHALS[0.59] TFSNRFI[0.40 ± 0.02] VVPTPPY[0.76 ± 0.06] ELAPDSP[0.59] 12 Phage display (common panning) 3 PMID:22363498 Anti-LPS monoclonal antibody Lipopolysaccharide, LPS Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA In phage ELISA, absorbance was measured at 490 nm. Each experiment was repeated three times. Data were reproduced from the graph and expressed as means ± SD. Phage 2Ab as negative control showed no ELISA signal. The data demonstrate the identification of synthetic peptides that mimic LPS by interacting with TLR-4. This LPS mimotope-TLR-4 interaction will allow for the development and use of these peptides as a new class of adjuvants, namely TLR-4 agonists. 1948 TLHLHHL(15) 1 Phage display (competitive panning) 5 PMID:22372912 Transcriptional regulatory protein devR (dosR) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the first round, the phage library was incubated with (His)6-DevR immobilized on Ni2+ NTA (nitrilotriacetic acid) magnetic agarose beads. The loosely bound phages were discarded by elution with DevR D54N mutant protein (100μg/mL concentration), which differs from the wild-type protein at the phosphorylation site. A second elution was performed with buffer containing 250 mM imidazole that released the His6-tagged protein along with the tightly bound phages. Three rounds of panning were performed using the twin elutions approach, and each time the phages in the imidazole elution were amplified and used as an input for the next round of panning. The fourth and fifth rounds were performed on plate to eliminate bead binders. The loosely bound phages were\r\nfirst eluted with mutant D54N DevR protein and then with 0.2 M glycine, pH 2.2. In the fifth (final) round of panning, a penultimate elution using phosphorylated DevS was carried out prior to final elution of the bound phages using 0.2 M glycine. DevRS1 (TLHLHHL) inhibited DevR-regulated transcription and survival of nonreplicating tubercle bacilli in a hypoxia model of dormancy. DevRS1 peptide-mediated inhibition demonstrates the efficacy of intercepting DevR function to block hypoxic adaptation of M. tb. 1949 HTPPVTS(5) ASTLPKA(3) QPQVPDA(3) ALTPTPP(2) ILGVGLP(2) SILPYPY(2) LGSTTPP(1) SPIWMHS(1) SPLLSTP(1) AXQISPP(1) TAPTSPS(1) QPLELPN(1) AAQTSTP(1) FSAHAHL(1) NQDVPLF(1) TPRLLVE(1) HAIYPRH(1) IPTLPSS(1) EATTRAY(1) YSIPKSS(1) ATPLWLK(1) FPPNKES(1) LPSYHVP(1) YRAPWPP(1) DTWRPVR(1) THPLLLS(1) SPQMTLS(1) SSHTISF(1) HPFEHFS(1) STATSKT(1) MLQPQAP(1) WGGLPEP(1) EPLVQHP(1) 33 Phage display (subtractive panning) 3 PMID:22404231 Thiamethoxam, TMX Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) One hundred microliters of the diluted phage library was then introduced into the TMX suspension and incubated for 1 h with gentle rocking. The sample was washed several times with TMX-saturated 0.1% TBST buffer and transferred to new centrifuge tubes. This step removed nonspecific binding phage or any phage with a strong affinity to the polypropylene centrifuge tubes. The six most strongly binding phages exhibit at least 1000 times the binding affinity of wild-type M13 and express heptapeptide sequences that are rich in hydrophobic, hydrogen-bonding amino acids and proline. Among the peptide sequences identified, M13 displaying the pIII domain heptapeptide ASTLPKA exhibits the strongest binding to thiamethoxam in competitive binding assays. Electron and confocal microscopy confirm the specific binding affinity of ASTLPKA to thiamethoxam. 1950 FPMFNHWEQWPP(3) 1 Phage display (common panning) 4 PMID:22421431 Epidermal growth factor receptor Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After the fourth round of panning, seven phage clones from the 12-mer peptide displayed phage library were randomly selected and used for ELISA to rhEGFR. Three of the seven clones from the 12-mer peptide library showed a specific affinity to rhEGFR, and all three encoded the same 12 amino acid peptide sequence, FPMFNHWEQWPP. 1951 HYDRHNYHWWHT(7) LSKNLPLTALGN(1) SGMKEPELRSYS(1) 3 Phage display (common panning) 3 PMID:22424885 Anti-SapA monoclonal antibody A2D5 S-layer protein Not determined. Not determined. Ph.D.-12 phage display library (X12) 15 phage clones were randomly chosen and tested for reactivity with mAb A2D5 by indirect ELISA. Single-stranded DNA from positive clones was sequenced and compared with the sequence of SapA to predict the key epitope. ELISA and/or Western blot analyses further validated that synthetic peptides and recombinant peptide mimotopes all interact with mAb A2D5. Nine of ten positive phage clones identified by screening were sequenced successfully. Seven clones shared the same sequence HYDRHNYHWWHT; one had the sequence LSKNLPLTALGN; and the final one had the sequence SGMKEPELRSYS. These three sequences shared high homology with SapA J05577 in the region GNEKDFVTKIYSIALGNTSDVDGINYW, in which the underlined amino acids may serve as key residues in the epitope. 1952 CPIKTLPMC(18) 1 Phage display (common panning) 4 PMID:22426388 Beta-amyloid protein 42 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA To confirm further the binding of XD4 (CPIKTLPMC) to Aβ42, Aβ42 or XD4 at different concentrations was coated onto ELISA plates. Subsequently, the phages expressing XD4 peptide, His tag conjugated XD4, or Aβ42 were added to the wells for ELISA measurement. The results showed that the phages and His tag-conjugated XD4 bound to Aβ42 in a dose-dependent manner. Conversely, Aβ42 also bound to coated XD4 in a concentration-dependent manner. Thirty individual positive clones binding to Aβ42 with higher phage ELISA values were selected and amplified. The DNA from these 30 positive clones was extracted and sequenced. The level of consensus of the sequences was examined. The results indicated that 18 clones possessed the same sequence (PIKTLPM, denoted as XD4). XD4 significantly inhibited Aβ cytotoxicity, increased the microglial phagocytosis of Aβ, decreased the Aβ-induced generation of ROS and NO, and attenuated the disequilibrium of calcium homeostasis in vitro. Remarkably, XD4 also attenuated memory deficits in β-amyloid precursor protein/presenilin 1 (APPswe/PS1dE9) transgenic mice, and reduced amyloid plaque burden and Aβ40/42 levels. 1953 EMPWSTNSFTDD(1) THQNNTSSLLGF(1) ARLHMASTASSL(1) YSDWQASTSSRL(1) HQSISPSTSSKL(1) THQHSTNSYLLL(1) IMNSTSSFSHFQ(1) AESTSSFKSPHH(1) YASMLHSTSSKL(1) SAGPSTSSELSS(1) DYNRLNSTSSAL(1) ERPSAVNTSSWL(1) GYMEDANTTSKL(1) DMSLSSSTSSLL(1) RADMPSTSSALS(1) FTNTTSFLVART(1) TLTYAPSTSSPL(1) STKLNSTSSTLG(1) HKNLSNSTASLL(1) ENHTHSNTSSAL(1) LNSTVSFLQRSP(1) SMYSTSSTLWAD(1) THRYNSTSSLLT(1) QYSTSSSLGISI(1) DSRPSPSTSSHL(1) YPSDWTSTSSML(1) SPTHALSTSSQL(1) VPIEAKSTSSFL(1) FTWNHASTASFA(1) FMTIEASTASSL(1) ESLTNNSTASHL(1) VSPHMMSTASAL(1) QGWSTSSFTTRP(1) SNQVVHQHSTLS(1) TLNQHNTSSSLG(1) SPPARSSTSSQL(1) TSGQSTASHLLL(1) IQPYASTSSALW(1) TSPHGQSTSSFT(1) 39 Phage display (common panning) 3 PMID:22427942 Anti-MSP1a monoclonal antibody 15D2 Major surface protein 1a, MSP1A Not determined. Not determined. Ph.D.-12 phage display library (X12) The consensus sequence SxSSQSEASTSSQLGA was obtained, which is located in C-terminal end of the 28-aa repetitive motif of the MSP1a protein, but the alignment and sequences' variation among mimotopes allowed us to map the critical motif STSSxL within the consensus sequence. 1954 ATKVKIPFEAKV VATPVPPTLTPF ATLRTYPYMDRA QLAPMATHDKHP YALRPGMPQWLE TPPTYSWFTHRM GSATNPTMGQRM AETHVLNKHTPL HSSSHWSWSTPL NMRLLANPAMAG QIPAQNRLVFLT VPGWDSHNARHQ HAESPFPNPTRA NKITLHSNSSIA 14 Phage display (common panning) 3 PMID:22428072 Human corneal epithelial cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Fourteen sequences were obtained and BLASTp analysis showed that most of their homologue counterparts in GenBank were for defined or putative proteins in various pathogens. 1955 RKRIRRMMPRPS(5) TNLKSIRRPQIP(2) LRKRNNPIRTKM(1) SSTRMISIRQST(1) KMPRQSNRRLMM(1) RHLQTSPIMLQI(1) RHTNSTRLNRPT(1) 7 Phage display (common panning) 3 PMID:22459587 Thiacloprid Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1956 RNRHTHLRTRPR(6) RRPKRKHRMHLS(2) ISKHTPLRTRPR(1) RRPKRQHTMQLS(1) SMRMRIHIPSSH(1) RQIIQPSPKQRR(1) 6 Phage display (common panning) 3 PMID:22459587 Imidacloprid Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1957 YNTNHVPLSPKY(20) 1 Phage display (subtractive panning) 7 PMID:21209841 Extracellular domain of human carbonic anhydrase IX Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Immobilized recombinant extracellular domain of the epidermal growth factor receptor (EGFR) was used for negative selection. After 7 selection rounds, single-stranded DNA from isolated bound phages was sequenced. All 20 clones sequenced, displayed the same peptide sequence: YNTNHVPLSPKY (CaIX-P1). In vitro binding experiments of 125I-labeled-CaIX-P1 revealed an increased uptake of the radioligand in the CAIX positive renal cell carcinoma cell line SKRC 52. Binding of the radioligand in the colorectal carcinoma cell line HCT 116 increased with increasing cell density and correlated with the mRNA expression of CAIX. Radioligand uptake was inhibited up to 90% by the unlabeled CaIX-P1 peptide, but not by the negative control peptide octreotide at the same concentration. No binding was demonstrated in CAIX negative CaKi 2 and HUVEC cells. Organ distribution studies revealed a higher accumulation in SKRC 52 tumors than in heart, spleen, liver, muscle, intestinum and brain, but a lower uptake compared to blood and kidney. These data indicate that CaIX-P1 is a promising candidate for the development of new ligands targeting human carbonic anhydrase IX. 1958 VDQNAKSWGQMK(1) QSYSAKMWGQVG(1) STDIISWKHWGS(2) EHWPQWGLALKR(2) TNLETLKQWGFP(1) GNPAPWKDWGSA(3) QSDKHWGMATPP(1) DFYSWKHWGSTT(1) YDNKEWGIARPN(1) TPPPTYKLWGAL(1) SSSKTWGAPLYY(1) SSNDTKAWGFAV(1) DYRSWGAVPLQL(1) NSSSNKHWGSVP(1) KXWG 14 Phage display (common panning) 3 PMID:21233208 Anti-D2NS1 monoclonal antibody DB16-1 Non-structural protein 1, NS1 Not determined. Not determined. Ph.D.-12 phage display library (X12) The B-cell epitope of DB16-1 displayed a consensus motif, Lys-X-Trp-Gly (KXWG), which corresponded to amino acid residues 116-119 of DV NS1 and mimicked amino acid residues 334-337 in LYRIC. Moreover, the binding activity of DB16-1 in NS1 of DV-2 and in LYRIC disappeared after the KXWG epitope was deleted in each. In conclusion, DB16-1 targeted the same epitope in DV NS1 and LYRIC protein on human endothelial cells, suggesting that it might play a role in the pathogenesis of DHF/DSS. 1959 APSPMIW(5) LQNAPRS(3) HAIYPRH(1) 3 Phage display (common panning) 3 PMID:18931708 CD133 extracellular fraction Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the screening, the phage display peptide library was subjected to three rounds of affinity selection. Each affinity selection was done by biopanning on ELISA plate. ELISA plates were blocked with bicarbonate buffer (PH 8.6) containing 50 mg/l streptavidin. 1960 LVGVFH(0.8) 1 Phage display (common panning) 3 PMID:22452335 Porcine ear skin Not determined. Not determined. Not determined. f3-6mer phage display library (X6) Skin was sandwiched between the donor and receptor compartments with the SC facing the donor compartment. Phages, from phage display peptide library (PDL) were loaded into the donor compartment of the Franz diffusion cell. The donor compartment was covered with parafilm to maintain occlusive conditions. Phage particles that crossed the skin were collected from the receptor chamber after 24 h and were amplified. An amplified pool of phages was loaded again in the donor chamber for another round of screening. In this way, screening was performed three times. Eighty percent of the phages that consistently crossed the porcine skin possessed T2 peptide (LVGVFH) on their surface. Pretreating the skin with synthetic T2 peptide at pH 4.5 resulted in significant penetration enhancement of hydrophilic drug 5-fluorouracil (5-FU) across skin. FTIR spectroscopy showed that the T2 peptide interacted with skin lipids to enhance the skin penetration. Pretreating the skin with T2 peptide enhanced the partitioning of small molecules with different lipophilicities (5-FU, fluorescein isothiocyanate, and rhodamine 123 hydrochloride) into skin. Fluorescence studies showed that T2 peptide enhanced the diffusion of these molecules into intercellular lipids of SC and thus enhanced the penetration into the skin. Histidine at the c-terminus of T2 peptide was identified to be critical for the skin penetration enhancement. T2 peptide interacted with skin lipids to cause skin penetration enhancement. 1961 GEVGEQEKARVG(1) EGKGVEAVGDGR(2) AEPDATGWRSLG(1) 3 Phage display (common panning) 3 PMID:22484444 Chitin Not determined. Not determined. Not determined. SUT12 M13 phage display library Based on phage ELISA results, Four phage clones showed binding signals two times higher than the background. The synthetic chitin binding peptide (ChiBP) could bind to chitin beads and disrupt their structure. This selected peptide was successfully used to immobilize alkaline phosphatase on chitin. In addition, the peptide could induce colloidal chitin in water to form a chitin coat on the surface of plastic tubes. Scanning electron microscopy (SEM) revealed that the peptide could induce colloidal chitin and chitohexaose to form networks when the temperature was raised to 42 ℃. 1962 AETVESCLAKSH(1) ALNWASKSSGRY(2) ANYFLPPVLSSS(2) HPVLHLPISTPR(1) ILPVQGIAMRSM(1) LTPTVRSGYLNH(1) MKPGQLRTGSTM(1) MTEGMNRSQLPA(1) NDSSSALPHTLT(1) NHVHRMHATPAY(2) NIYSTELAAPSP(1) NLILTPGWSRLA(1) NMFGSLTSHVTA(1) NWELQSTPSHTA(1) SKHFNPGLATAD(1) SQLFITCPPISL(1) SQLSITSPPISL(3) TAMTQRISAGNH(2) VATVMTTSAFEP(1) WHWNLWAPASPT(1) 26 Phage display (common panning) 3 PMID:22515661 Type IV secretion system protein virB8 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The authors have pursued an unbiased approach using peptide arrays and phage display to identify interaction partners and interaction domains of type IV secretion system assembly factor VirB8. These approaches identified the globular domain from the VirB5 protein to interact with VirB8. This interaction was confirmed in cross-linking, pull-down, and fluorescence resonance energy transfer (FRET)-based interaction assays. In addition, using phage display analysis, the authors identified different regions of VirB6 as potential interaction partners of VirB8. 1963 CLPSTNAMC CLSTLRSTC CNRLILPHC CTTAQLRWC CLHRHSQLC CNPQPVAAC CWQNKALAC CYQTISAAC CTSMKAAIC CSLGSTIVC CKFTLSLSC CRHALNTAC CLSAGGAEC CSLGSTIVC CYQTISAAC CTILSGSRC CLSAGGAEC CKFTLSLSC CTSMKAAIC CYQTISAAC CLSAGGAEC CLSTLRSTC CSLTALHRC CTTAQLRWC CKLLYYVSC CPGLSAGRC CLPSTNAMC CLTAWGWAC CSLGSTIVC 29 Phage display (common panning) 3 PMID:22515661 Type IV secretion system protein virB8 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The authors have pursued an unbiased approach using peptide arrays and phage display to identify interaction partners and interaction domains of type IV secretion system assembly factor VirB8. These approaches identified the globular domain from the VirB5 protein to interact with VirB8. This interaction was confirmed in cross-linking, pull-down, and fluorescence resonance energy transfer (FRET)-based interaction assays. In addition, using phage display analysis, the authors identified different regions of VirB6 as potential interaction partners of VirB8. 1964 WAETWPLAQRPP(1) DFRWATHMHTPA(6) GTPPMSPQVSRV(1) HFRWANHTHFQT(1) TSQYQSPRAVHP(1) HFRWANHTHFQT(1) GLRNSVPYQTFT(1) HFRWANHTHFQT(1) GHGLLQYTDVMF(1) QHTYWPNYTPLL(1) HTRRTTHHILR(1) APIKAPTIRDTA(1) SKFEPISKYLQP(1) 18 Phage display (common panning) 3 PMID:22536790 Major prion protein, PrP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Lyophilized streptavidin 1.5 mg (Ph.D.-12, BioLabs, New England) was dissolved in 1 mL of 10 mM sodium phosphate (pH 7.2) in 100 mM NaCl. For each target three wells of a 96-well plate were coated with the prepared solutions, i.e., 150μL/well overnight at 4°C under gentle agitation. The prion-targeting peptide P9 (WAETWPLAQRPP) shows homology to prion protein aa 16-28, i.e., WSDLGLCKKRPK. Prion-targeting peptide P50 (DFRWATHMHTPA) matches the identified prion consensus sequence as described in the text (X1FRWAX6HX8HX10X11X12), showing homology to prion protein aa 147?155, i.e., DRYYRENMHRYP. 1965 HLNILSTLWKYR(1) 1 Phage display (common panning) 3 PMID:22536790 Ganglioside GM1 Cholera toxin B protein, CTB Not determined. Not determined. Ph.D.-12 phage display library (X12) Lyophilized streptavidin 1.5 mg (Ph.D.-12, BioLabs, New England) was dissolved in 1 mL of 10 mM sodium phosphate (pH 7.2) in 100 mM NaCl. For each target three wells of a 96-well plate were coated with the prepared solutions, i.e., 150μL/well overnight at 4°C under gentle agitation. The authors show that the prion-targeting peptides do not induce efficient transcytosis of polymersomes across the BBB in vitro nor induce accumulation of polymersomes in the brain in vivo. In contrast, the G23 peptide is shown to have transcytotic capacity in brain endothelial cells in vitro, as well as a brain-targeting potential in vivo, as reflected by the accumulation of G23-polymersomes in the brain in vivo at a level comparable to that of RI7217-polymersomes, which are targeted toward the transferrin receptor. Thus the G23 peptide seems to serve both of the requirements that are needed for efficient brain drug delivery of nanocarriers. An unexpected finding was the efficient accumulation of G23-polymersomes in lung. In conclusion, because of its combined brain-targeting and transcytotic capacity, the G23 peptide could be useful in the development of targeted nanocarriers for drug delivery into the brain, but appears especially attractive for specific drug delivery to the lung. 1966 AADNAKTKSFPV(12) IVWPTSPRALDA(2) 2 Phage display (subtractive panning) 3 PMID:22563192 Human gastric cancer specimens Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) For screening, non-specific binding phage was cleared from the library by panning against normal appearing gastric mucosa adjacent to the tumor. Approximately 92.8% of the non-specific phage clones were subtracted from the original phage library after two rounds of biopanning against normal appearing gastric mucosa. The values for AAD peptide were statistically significant (P < 0.01) for gastric cancer as compared with other histological classifications and control peptide. The peptide AAD identified in this study has the potential to guide tissue biopsy and improve the detection of pre-cancerous lesions in gastric mucosa. 1967 CYWELEWAPC 1 Phage display (common panning) 3 PMID:22578098 Cardiac phospholamban, PLB Sarcoplasmic/endoplasmic reticulum calcium ATPase 2, SERCA2, SR Ca(2+)-ATPase 2 Not determined. Not determined. CX8C fUSE5 phage display library Panning steps of the primary library were performed on 10-mm multiwell plates coated with streptavidin essentially. The target was the 36-PLB-C peptide biotinylated on the cysteine residue. Three enrichment cycles (e12 phage particles used in the first round, then 20% of the volume of the amplified eluates were used in the second and third rounds) were carried out. Its functional activity was tested in Ca2+ uptake assays utilizing preparations from cardiac sarcoplasmic reticulum. By synthesizing and testing a series of alanine point-mutated cyclic peptides, the authors identified which amino acid was important for the inhibition of the phospholamban function. The structures of active and inactive alanine-mutated cyclic peptides, and of phospholamban (1-36), were determined by NMR. This structure-activity analysis allowed building a model of phospholamban-cyclic peptide complex. 1968 SLTNLSK(16) 1 Phage display (subtractive panning) 4 PMID:22588916 Human osteosarcoma MG-63 cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Human embryonic kidney 293T cells was used to carry out subtractive panning. The sequence SLTNLSK was confirmed as the most frequent peptide by DNA sequencing and showed strong specificity verified by cell ELISA, fluorescent staining and organ immunohistochemistry. 1969 HLPLFRHVWPNM(1) HLSTWLSHSWLR(1) HLRQTLWNLSTS(1) HWWQWPSSLQLR(1) HNWTRWLLHPDR(1) HWFSAWYQLGSN(1) HWKPWWFSPGSW(1) HWWHFYQPPLST(1) HFSWFLPWNAPL(1) HFTWWYPGYNTP(1) HSWFWQAWPPQL(1) HIRLPTWWGAYP(1) FHENWPSGGGSA(1) FHKNWPIWWYPP(1) WHNPYQLWSWRW(1) ASQVGPAHTSTV(1) SDRIPSLRAHTV(1) ITKSGLTHLYPV(1) GPQVLANIPHFP(1) 19 Phage display (common panning) 3 PMID:22606046 72 kDa type IV collagenase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The authors showed that M204C4 (HWWQWPSSLQLRGGGS) and M205C4 (HNWTRWLLHPDRGGGS) inhibited the activity of MMP-2 in a dose dependent manner in vitro. Two peptides reduced MMP-2 mediated invasion of the pancreatic cancer cell lines PANC-1 and CFPAC-1, but not affected the expression and release of MMP-2. Furthermore, these two peptides could suppress tumor growth in vivo. 1970 CTATSHLFC(4) CGNSWYNSC(3) 2 Phage display (subtractive panning) 4 PMID:22653674 IgGs purified from equine sera Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Phages library were added to the microtiter well adsorbed with negative IgG firstly. 1971 TTSVLLHCIEWTILNQ 1 Phage display (common panning) 4 PMID:22683611 Caspase-6, CASP-6 Not determined. Not determined. Not determined. Linear-lib M13 phage display library In each round, biotinylated zymogen caspase-6 was incubated with the amplified phage from previous round and the caspase-6-phage complex was captured by NeutrAvidin-coated plate that has been previously blocked by block buffer. After four rounds of binding selection, individual phage clones were analyzed in a high-throughput phage spot ELISA using plate-immobilized zymogen caspase-6 as the target. The binding signal of the same phage particle to NeutrAvidin was detected as non-specific binding noise. Clones with phage-binding signal to target over 0.5 and signal/noise ratio > 3 were considered to be positive clones and were subjected to DNA sequence analysis. 1972 VFVVCDWFDFVCALGM VFVVCDWFEFVCTFGL VIVSCDWVYFICELVM 3 Phage display (common panning) 4 PMID:22683611 Caspase-6, CASP-6 Not determined. Not determined. Not determined. Cyclic-lib M13 phage display library In each round, biotinylated zymogen caspase-6 was incubated with the amplified phage from previous round and the caspase-6-phage complex was captured by NeutrAvidin-coated plate that has been previously blocked by block buffer. After four rounds of binding selection, individual phage clones were analyzed in a high-throughput phage spot ELISA using plate-immobilized zymogen caspase-6 as the target. The binding signal of the same phage particle to NeutrAvidin was detected as non-specific binding noise. Clones with phage-binding signal to target over 0.5 and signal/noise ratio > 3 were considered to be positive clones and were subjected to DNA sequence analysis. 1973 FLSPAQRFELFP(4) WPSAAEAFSKSP(3) APSAADQFMALF(2) MPSAVHLFETMR(1) DNSAAQYFSRMP(1) NSAAYSFTLFPG(1) QSAASLFNNSMN(1) SAADRFRTGPMP(1) SAADRFSRYHER(1) GASAAENFLKSL(1) DSAASHFTTQFR(1) KSAAEQFSLTMA(1) SAANKFTVATMP(1) GASAVTRFQSMY(2) SAADHWRLSALI(1) YQSAASKWSTYY(1) SAASKWHNALAQ(1) SAVHQWATQFLM(1) SAVNNWESFYGW(1) SHTAAHHFQVML(1) SMTAATVFWTQM(1) YNTAATKFSLQL(1) STAVQQFQNTLS(1) VSTAVSRFEALT(1) AEPAAARWQASL(1) QPAAVRLFEMMT(1) EPNIADFFYRSM(1) ADQNSFISALFQ(1) TQNMHKFFEQES(1) AKITRTLSLPFS(2) QKITTTFFSPLM(1) AKLTSTFSESPH(1) SKLTSTLTQLAS(1) AHPITQHDTSAR(1) SPLLDLLKLRSP(1) SPQQQTLATFFG(1) APIQDNTLGRWF(1) TFPQNVPLPILP(1) EHEQHLSLQRWF(1) EELFTRNVLNWP(1) SIAEKWTTYLRK(1) KSLSRHDHIHHH(22) KSVSSQDHIHHQ(1) RMNSPLDVLHHR(1) SSLHTHQTPMFM(1) P-S-A-[AV]-X-R-[FW]-[ES]-L-S-P, A-K-[IL]-[TQ]-X-T-L-X-L, K-S-L-S-R-X-D-X-I 45 Phage display (subtractive panning) 4 PMID:22716192 Anti-MgPa polyclonal antibody Adhesin P1 Not determined. Not determined. Ph.D.-12 phage display library (X12) In each round, the phage eluates were added into the microtiter plate coated with normal rabbit antiserum at room temperature and incubated for 1 h. The preincubation procedures were performed 5 times to remove those phages that possibly nonspecifically bind to rabbit antiserum. Results of bioinformatics analysis by MIMOX demonstrated that the key consensus amino acid residues in the aligned mimotopes may be S, A, and F for cluster 1; A, K, I, T, and L for cluster 2; and K, S, L, R, D, and I for cluster 3. Three representative phages could recognize sera from M. genitalium-positive patients to varying degrees, whereas they could not recognize the sera from Mycoplasma pneumoniae-positive patients or the sera from healthy people. These findings will help to clarify the mimic epitopes of MgPa to facilitate diagnosis of the antigen and to understand the antigenic structure of MgPa. 1974 WDSDCKRSCRVH(1) LTWVSDSKSGNT(1) TIAPSWATDSKP(1) TPNNAQKQPQLP(1) SWMPDSKVFASH(1) WETDQKFKQRVA(2) SAYDDVKRFYTN(1) 7 Phage display (subtractive panning) 2 PMID:22720103 Anti-VSG LiTat 1.3 polyclonal antibody Variant surface glycoproteins LiTat 1.3, VSG LiTat 1.3 Not determined. Not determined. Ph.D.-12 phage display library (X12) A negative selection was conducted with endemic negative serum antibodies coated on magnetic particles. Eight of 188 phage clones reacted in the sandwich ELISA. 1975 ETDNMKPLHLRQ(1) VNDASKLFYPRS(1) WPTSWHMWLANR(1) GVPDNHKPARTQ(1) ALPTHMNWVMPV(1) TWPQWWWTNSKG(1) NPPIWGTATKGI(1) FWKPHRTHFWWG(1) YNWETDKPMPVP(1) TWWWHSLAKTPH(1) TTWNFKHWWPYR(1) 11 Phage display (subtractive panning) 3 PMID:22720103 Anti-VSG LiTat 1.3 polyclonal antibody Variant surface glycoproteins LiTat 1.3, VSG LiTat 1.3 Not determined. Not determined. Ph.D.-12 phage display library (X12) A negative selection was conducted with endemic negative serum antibodies coated on magnetic particles. Eleven of 188 phage clones reacted in the sandwich ELISA. 1976 AAIMHQEQESNT(1) 1 Phage display (subtractive panning) 1 PMID:22720103 Anti-VSG LiTat 1.5 polyclonal antibody Variant surface glycoproteins LiTat 1.5, VSG LiTat 1.5 Not determined. Not determined. Ph.D.-12 phage display library (X12) A negative selection was conducted with endemic negative serum antibodies coated on magnetic particles. One of 188 phage clones reacted in the sandwich ELISA. 1977 SAGFENDGTKLA(1) TGLPTTNKQTSS(1) 2 Phage display (subtractive panning) 2 PMID:22720103 Anti-VSG LiTat 1.5 polyclonal antibody Variant surface glycoproteins LiTat 1.5, VSG LiTat 1.5 Not determined. Not determined. Ph.D.-12 phage display library (X12) A negative selection was conducted with endemic negative serum antibodies coated on magnetic particles. Two of 188 phage clones reacted in the sandwich ELISA. 1978 AYSKPTIKLANP(3) LPLATADKNGRT(1) DKLDNPGGPTVG(2) LQMPHNSKTANP(1) INGQFSLKYRNP(1) LMPNKISNFASA(1) DQTCNSPPCPPL(1) STLPPPQGKIIH(1) WYPLHSGLRSYY(1) NKSTTNDFLRSP(1) NGDYLQYKAPNP(1) NTVRPPTLFYHW(1) WHSEYQEPYPLS(1) LDKNVLSPPMPL(1) HHMSWYSRWLPV(1) WWKPWSNFYGST(1) LNTQNHAPLPSI(1) 17 Phage display (subtractive panning) 3 PMID:22720103 Anti-VSG LiTat 1.5 polyclonal antibody Variant surface glycoproteins LiTat 1.5, VSG LiTat 1.5 Not determined. Not determined. Ph.D.-12 phage display library (X12) A negative selection was conducted with endemic negative serum antibodies coated on magnetic particles. Seventeen of 188 phage clones reacted in the sandwich ELISA. 1979 QWGWNMPLVEAQ LLADTTHHRPWT HSHLHIHSGIQA NHVHRMHATPAY HSSHHQPKGTNP HSSPHFSRTWAS HYQHNTHHPSRW HHRTLSPSVSIL HSSPHFSRHGLL HHGHSPTSPQVR NTIHHRHHMPPP SHNHPPRHTAHS SSGLRHSHHQHP HSKLNNRHHALL HTKPHHTPTQRA GHIHSMRHHRPT HS-X(2)-H 16 Phage display (common panning) 3 PMID:22720657 Oxygen-terminated sides of single-crystalline ZnO (000-1) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1980 ERSWTLDSALSM SNNDLSPLQTSH DSSNPIFWRPSS SILSTMSPHGAT SHALPLTWSTAA LLADTTHHRPWT HVSIHRTTHHEM MKPDKAIRLDLL HYPTAKFHAERL TKNMLSLPVGPG FNTGSQMHQKFP HSSHHQPKGTNP HHTHRVDVHQTR FGLTAPRSASIL APRLPQSLLPQL HHGHSPTSPQVR SHNHPPRHTAHS HSKLNNRHHALL HTKPHHTPTQRA 19 Phage display (common panning) 3 PMID:22720657 Zinc-terminated sides of single-crystalline ZnO (0001) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1981 RRRKRPIRRKLR(1) HIRLTLSRNKNT(1) TRTMTMNQNRRS(1) PQTNQTTMKMRM(1) IMPTKKIPPIIM(1) 5 Phage display (common panning) 4 PMID:22743126 Helicase, Hel Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1982 KKTPPIRIRHRP(1) NIPIKPRPRLMK(1) SPHIIRNHRLSK(1) HRILMRIRQMMT(1) RIIRKSQRSLMN(1) RQPRTPMTRLSR(1) IIRHRSMIITIT(1) KTRTMQMRNRMP(1) RMRSKRRKITTR(1) 19 Phage display (common panning) 4 PMID:22743126 RNA-directed RNA polymerase, Pol, RdRp Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) P9 (HRILMRIRQMMT) exerted the highest antiviral activity with an IC50 of 56 mM, and the minimum toxicity to cells. It was proved that p9 inhibited PRRSV replication in infected MARC-145 cells in a dose-dependent manner, and the amino acid sequence of HRILMRIR was important for antiviral activity of p9. Also, p9 could bind to the cell membrane and penetrated into cells. 1983 AGKGTPSLETTP(4) GPLPKTYAIPSS(2) AHQANFPSSSAI(1) DNVHTTLSQPST(1) FPSSSNHYPWAE(1) KLTTNPSSPLNY(1) TPRAELHFGQSS(1) 7 Phage display (in vivo) 3 PMID:20145035 Hepatocellular carcinoma cell lines BEL-7402 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Phage binding assay was performed by using cell-based ELISA. The absorbance at 450 nm was determined. Selectivity is calculated using the following formula: Selectivity = (ODS1-ODC1)/(ODS2-ODC2). Here, ODS1 and ODC1 represent the values of the binding to BEL-7402 cells by the selected phage and control phage, respectively; ODS2 and ODC2 represent the values of the binding to control cell line HL-7702 by the selected phage and control phage, respectively. The phages with selectivity above 2 are considered to be positive clones. Phage clones A54, A67, and B2 seemed to have the most specific binding ability in cell-based ELISA. It was found that the peptide sequences AGKGTPSLETTP, TPRAELHFGQSS and AHQANFPSSSAI were displayed on phage clone A54, B2 and A67, respectively. Tumor was raised by s.c. injection of 1.0e6 BEL-7402 cells per mouse in 200μL culture medium with 10% FCS into the flanks of the nude mice. When tumors reached a size of ∼1 cubic centimeter, phage library containing e11 colony-forming units (CFU) was injected through the tail veins of the mouse. After phage A54 (AGKGTPSLETTP) was injected i.v. into the xenograft-bearing mice for in vivo distribution, phage enrichment was found in tumor tissues compared with control phage C10 and normal liver tissues through phage titering and immunohistochemical staining. Next, the specific binding ability of synthesized peptide A54 was further confirmed by fluorescence microscopy, competition binding, and fluorescence-activated cell sorting assay. A54 and A54M (sequence AGKGTAALETTP) were synthesized and coupled to doxorubicin (DOX) to do the preliminary targeting therapy. After the treatment, the proliferation of liver cancer cells treated with A54-DOX was restrained significantly in vitro when compared with A54M-DOX-treated group. Reduction in tumor size and prolongation of long-term survival were also found in xenograft-bearing models compared with free DOX-treated group. In conclusion, the specific binding peptide A54, which was screened from phage display library, represents a promising approach for the development of novel target therapy strategies against hepatocellular carcinoma. 1984 WMPSDVDINDPQ QPKPAAEAASKK WYSSMSEDKRGW 3 Phage display (common panning) 4 PMID:22123020 Zirconia Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) DNA sequencing of the corresponding portion of Ф#17 (WMPSDVDINDPQGGGSRPNLHQPKPAAEAASKKKSENRKVPFYSHSWYSSMSEDKRGW) unexpectedly revealed that it displayed a 58-mer peptide. The authors found that Ф#17 had a 300-fold, significantly higher binding affinity for zirconia discs than phages displaying no peptide. In quartz crystal microbalance assay, a rapid increase in energy dissipation was observed from Ф#17 but not from the control phages, indicating that Ф#17 binds to the surface of zirconia via its displayed peptide. 1985 YITPYAHLRGGN(14) 1 Phage display (common panning) 5 PMID:22435500 Amorphous spherical silica particles (15 nm diameter) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1986 KSLSRHDHIHHH(20) 1 Phage display (common panning) 5 PMID:22435500 Amorphous spherical silica particles (82 nm diameter) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1987 LDHSLHS(16) 1 Phage display (common panning) 5 PMID:22435500 Amorphous spherical silica particles (82 nm diameter) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 1988 MHRSDLMSAAVR(20) 1 Phage display (common panning) 5 PMID:22435500 Amorphous spherical silica particles (450 nm diameter) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 1989 NSWTNASLSTFH NSRTNNSQWTFQ ESWTNSWAHYFG ESWTNSWAMYFG QSYTNDDVLRIS QNMNNWTLASIM EVMNNWTLASIM ASISNLTLSRFM HSWSNYWGHQHA HRISNYAMELHS HSLTNTQMTQLS HSLSNIQMATLA HRMTNAMHHFMG HRMTNNAMDVFM HRLTNSEQAALP TAVTNSMMERLW GWGNKTPSQDVH DYTNSVSMRYLS HQLSNKDEQTPQ ADPFSPTNRIPL HSWTNSWMATFL 20 Phage display (subtractive panning, competitive panning) 3 PMID:22508944 Anti-MDA-LDL monoclonal antibody LR04 Malondialdehyde-modified LDL (MDA-LDL) Not determined. Not determined. Ph.D.-12 phage display library (X12) Control IgM was used in the negative selection, and then unbound phages were transferred to wells coated with LRO4 (positive selection). In the last biopanning round, LRO4-bound phages were competitively eluted with increasing concentrations (3-150 μg/ml) of MDA-LDL diluted in blocking buffer, followed by elution using elution buffer. Dodecamer peptide sequences were aligned by the Clustal W program to obtain consensus sequences, a dodecamer linear peptide P1 (HSWTNSWMATFL). 1990 CNNWNMPLC CNNRNMPLC CNNYNMPLC CNNQNMPLC CNNWKMPLC CNNSHMPLC CKNSXQPLC CNNSXMPLC CQNSHMPLC CNNSNMPLC CNNSKMRLC CDWAPHFTC CNNSNMPLC 12 Phage display (subtractive panning, competitive panning) 3 PMID:22508944 Anti-MDA-LDL monoclonal antibody LR04 Malondialdehyde-modified LDL (MDA-LDL) Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Control IgM was used in the negative selection, and then unbound phages were transferred to wells coated with LRO4 (positive selection). In the last biopanning round, LRO4-bound phages were competitively eluted with increasing concentrations (3-150 μg/ml) of MDA-LDL diluted in blocking buffer, followed by elution using elution buffer. Heptamer peptide sequences were aligned by the Clustal W program to obtain consensus sequences, a cysteine-constrained heptamer cyclic peptide P2(CNNSNMPLC). 1991 CTSYNEPLC CNLYNEPLC CNLYNEPFC CTVFNEPFC C-x(2)-[YF]-N-E-P-[LF]-C 4 Phage display (common panning) 4 PMID:23006741 Anti-VACV A33 monoclonal antidbody Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) MAb-1G10 binding peptides were isolated from the conformational library screening, none of which contain vaccinia virus A33 sequence.One consensus motifs were identified:C-x(2)-[YF]-N-E-P-[LF]-C. 1992 CQLKWPFEC CKPTWPFEC CSNSWPHEC CCDDWPHEC CQTSYPYEC CWYDSLIFC C-x(3)-W-P-[FH]-E-C 6 Phage display (common panning) 4 PMID:23006741 Anti-VACV A33 monoclonal antidbody Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) MAb-1G10 binding peptides were isolated from the conformational library scree-ning, none of which contain vaccinia virus A33 sequence. One consensus motifs were identified: C-x(3)-W-P-[FH]-E-C. 1993 MKPALCEPLCGM(5) NPYCEPVCQDWA(1) SLMRELCEPRCE(1) QLTAQHRDSLSS(1) C-E-P-L-C 4 Phage display (common panning) 4 PMID:23006741 Anti-VACV A33 monoclonal antidbody Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The conformationally constrained CEPLC sequence was likely to be functionally identical to the minimal core MAb-1G10 epitope. 1994 CNSHNHHTC[0.055] CHHNLTHAC[0.248] CLKQLQRGC[0.143] CLHHYHGSC[0.330] CTLYHNAGC[NG] 5 Phage display (common panning) 4 PMID:22759068 Protein tyrosine phosphatases(PTPRJ-His6) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA In phage ELISA, ELISA absorbance values of single phage clones were expressed as the difference between OD405 nm and OD629 nm. NG denotes not given. 1995 WPPKAMTQLGIKAC CGHGLKVQSTLGAC CKPYPKVYGLTGMC CQKLAWLTGKKEKC CTSKPQRWFGLPC CPPKLEQWYDGC 6 Phage display (common panning) 3 PMID:11179221 G protein β1γ1 subunits Not determined. Not determined. Not determined. f88-4 phage display library pool 1996 CEKRYGIEFC CEKRLGVRSC E-K-R-x-G-x(3) 2 Phage display (common panning) 3 PMID:11179221 G protein β1γ1 subunits Not determined. Not determined. Not determined. f88-4 phage display library pool 1997 YPWLQSY(6) LSTTLPL(1) 2 Phage display (subtractive panning) 3 PMID:22828501 Lens epithelium-derived growth factor (LEDGF)/p75 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) We first depleted the M13-derived linear 7-mer peptide phage librariy by preabsorbing the library on beads coated with wild-type HIV-1 IN.Phages remaining in the supernatant were used for positive selection rounds with LEDGF-coated beads . 1998 CVMGHPLWC(4) CILGHSDWC(3) CVSGHPLWC(2) CVEGHPAWC(1) CVSGHPEFC(1) CVMGHPTWC(1) CVMGHPSWC(1) V-x-G-H-P-x-W 7 Phage display (subtractive panning) 3 PMID:22828501 Lens epithelium-derived growth factor (LEDGF)/p75 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) We first depleted the M13-derived cyclic 7-mer peptide phage librariy by preabsorbing the library on beads coated with wild-type HIV-1 IN. Phages remaining in the supernatant were used for positive selection rounds with LEDGF-coated beads. Several of the isolated cyclic peptides(CPs) have a common motif (VM/xGHPL/xW) without linear or conformational homology to IN. 1999 FPWMQSYGVGIN(30) 1 Phage display (subtractive panning) 3 PMID:22828501 Lens epithelium-derived growth factor (LEDGF)/p75 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) We first depleted the M13-derived 12-mer peptide phage librariy by preabsorbing the library on beads coated with wild-type HIV-1 IN. Phages remaining in the supernatant were used for positive selection rounds with LEDGF-coated beads. 2000 IPLPPPSRPFFK 1 Phage display (subtractive panning) 4 PMID:22791264 Platelet-derived growth factor receptor β (PDGFRβ) Platelet-derived growth factor subunit B 3MJG, Not determined. Ph.D.-12 phage display library (X12) e11 plaque-forming units were added on immobilized negative target (EGFR) in 96-well plates. After incubation for 1 h at room temperature, medium containing unbound phages was transferred on the immobilized positive target (PDGFRβ) and further incubated for 1 h at room temperature. 40% of the clones isolated on immobilized protein displayed the peptide sequence: IPLPPPSRPFFK. 2001 SVSVGRKPSPRP NHRHYAMARNQS SFKPSGLPAQSL LSPHRSPQSPYS QRLNDHENPGHT HQMQSLSSNAST NNSQKPAPVSPF HSSLKLPNLSHR 8 Phage display (in vivo) 2 PMID:22960048 Mouse gut Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Animals underwent injection of the phage library in 200 ul of normal saline via tail vein at 4h following 30% TBSA burn injury and were compared to sham. One hour following injection of the phage library, animals were placed under general anesthesia with isoflurane and a segment of the distal ileum was harvested. 2002 SGHQLLLNKMPN(2) QQMHLMSYAPGP(1) QRLNDHENPVHT(1) NNHTKSQNITIS(1) SGHQLLLNKMPK(1) SHPWNAQRELSV(1) SIPSVAQSRSAT(1) 7 Phage display (in vivo) 3 PMID:22960048 Mouse gut Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Animals underwent injection of the phage library in 200 ul of normal saline via tail vein at 4h following 30% TBSA burn injury and were compared to sham. One hour following injection of the phage library, animals were placed under general anesthesia with isoflurane and a segment of the distal ileum was harvested. 2003 AKGHAETFSFSG(1) GDFNSGHHTTTR(1) SFKPSGLPAQSL(1) SGHQLLLNKMPN(1) DQRPLLTAGNPL(1) ILANDLTAPGPR(2) ISRPAPISVDMK(1) SDNVHTWQAMFK(1) 8 Phage display (in vivo) 4 PMID:22960048 Mouse gut Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Animals underwent injection of the phage library in 200 ul of normal saline via tail vein at 4h following 30% TBSA burn injury and were compared to sham. One hour following injection of the phage library, animals were placed under general anesthesia with isoflurane and a segment of the distal ileum was harvested. 2004 CSKAYLLLGQTC CKRTKAQILLAPC CTLRGKPYSLLGIC SIRKALNILGYPDYD CRKTTWILGEPLKC CEQTKTDRLLGNAC CEKASKILGVC K-A-x(2)-[IL]-L-G 7 Phage display (common panning) 3 PMID:11179221 G protein β1γ1 subunits Not determined. Not determined. Not determined. f88-4 phage display library pool The libraries were incubated with b-βγ subunites that had been bound to immobilized streptavidin on a microtiter plate. 2005 HPTTNTAHFFIT(1) SPVEALLTHRLH(1) NPIEQMLDYAKS(1) DHYAPERRPFFQ(1) SPVESRLHSNSS(1) NMVERMIERPSR(1) DHYAPERRPFFQ(1) HPTTNTADFFIT(1) NPVEMWSEMQHI(1) DHYAPERRPFFQ(1) TAHPTYKPPSPN(2) YDNGFSNSHMPL(1) QPLEHASHLTDV(1) HIDGTSLTLKKT(1) KAFAGTHTDTIT(1) 15 Phage display (subtractive panning) 5 PMID:22796092 Human immunoglobulin G (hIgG) Immunoglobulin G-binding protein A Not determined. Not determined. Ph.D.-12 phage display library (X12) To screen the positive or specifically binding phages,phage solution were added to the hIgG-coated beads.Before proceeding to the next round of biopanning,non-specific phages that were included in the selected phages were removed by a 'subtraction' step by using magnetic particles without a hIgG coating.The supernatant from this subtraction step was introduced to the next round. 2006 HSACDLPMHPMC(6) HSACDLLMHPMC(1) HSACDLPKAPWC(1) 3 Phage display (common panning) 6 PMID:22821302 CD59 T-cell surface antigen CD2 Not determined. Not determined. Ph.D.-12 phage display library (X12) All of these three screened amino acid sequences had nine hydrophobic residues. 2007 LEYTWGNHPOSR LEYTWRNHPOSR FRYTWGNHPHRG LRYTWGNWPLTA HNLEYTWGNWPM QHDSTYPWRNHP DMWMNHESYSNN PRLPFLHLTLEY TSLDKYTELQDY HDMRHRHMYOIA [LF]-[ER]-Y-T-W-[GR]-N-[HW]-P 10 Phage display (common panning) PMID:22824180 Anti-Bla g 2 monoclonal antibody 7C11 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2008 RIRALFGRSPVPCCV(2) MRRPAPWLASRLMRP(1) GRLRYTSHSARIQRV(2) RSALRCLARVESCRQ(1) ARHQRFLSSIQRAPF(1) ALVRHPGARLRSFTA(1) ARFRHSTKSAQFVPL(1) GRLRPAKNHSVTVRR(1) PRRHGFSPSVRAVLP(1) RVPPRYHAKISPMVK(13) RRPHSSHSHVSRFTS(1) R-x(2)-A-x-[FW]-x(2)-S-x-[VL] 11 Phage display (common panning) 5 PMID:23000297 Trisialogangliosides (GT1b) Tetanus toxin Not determined. Not determined. X15 fUSE5 phage display library The consensus motif(R-x(2)-A-x-[FW]-x(2)-S-x-[VL]) of the GT1b-binding peptides had one cationic amino acid,one hydrophilic amino acid, and multiple hydrophobic amino acids. Carbohydrate recognition in which the amino acids in the motif were considered to be involved was suggested by the combi-nation of hydrophobic interaction and hydrogen bonds as well as glycan-binding proteins. 2009 GGHRPRFSGSFVASRA(1) GVNRALPARWELWYPR(1) GHGLNAHLRPRPFLAR(1) GRNWNWPLRARVLSDA(1) GGRFHARPRTSSVWSF(1) GGTYYKRFGHSIPLVG(4) GPRRHRFSPSVRAVLP(2) GPRRGHFDESRFVHAV(1) GSSFSSGFVNWHRFAA(1) GYWPIDHKRVPLSRLV(1) GASRSRFTSHWKRRTI(1) GHRTQLNRLRSHRSVV(1) GGGRFPHARDRVSFSR(2) G-x(3)-[RK]-x(2)-G-x(2)-[VI], N-x(3)-P-x-R-x(2)-[LV] 13 Phage display (common panning) 5 PMID:23000297 Ganglioside GM2 Not determined. Not determined. Not determined. X15 fUSE5 phage display library 2010 DFRRLPGAFWQLRQP(4) GWWYKGRARPVSAVA(15) RWGALLRGGAAALFQ(1) W-x(4)-R-x(4)-[SA] 3 Phage display (common panning) 5 PMID:23000297 Ganglioside GD1a Not determined. Not determined. Not determined. X15 fUSE5 phage display library 2011 FAPQTTFTRPPY FTTPTTFARPQY SGHQPMLNKMPN FRSFESCLAKSH FTTPTTFARPPY YGHQRMLNKLPY FTTPNDFCSSSV FTTSLFDRDFCS GGYDLKRPLARP APEKSLIRNVHD SLSQHMLSSPSF TSSLVTARSILS LLPPDKSPRVFA ESKLPLPGAPEP 14 Phage display (common panning) 3 PMID:23000375 Interleukin-10 receptor subunit alpha (IL-10RA) Interleukin-10, IL-10 1J7V,1Y6K, Not determined. Ph.D.-12 phage display library (X12) Fourteen different peptides from low affinity to high affinity for functional characterization are selected. All the peptides were further ascertained for binding to shIL-10Ra by ELISA,of which peptide-04 (FRSFESCLAKSH) showed slowest dissociation. 2012 VPHNPGLISLQG(9) DPHNGRFPSTQL(3) VPKNLRFPSTSL(1) QHGKPGSIPLQG(1) LHVNPGLQTLQG(1) VPTNVQLHVLQG(1) VQHHPGIPDLQG(1) ARLHMYSTDVQS(1) VPHNPGLISLQG 8 Phage display (subtractive panning) 8 PMID:23017232 Staphylococcus aureus, S.aureus Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage library was twice depleted of clones binding to E.coli ATCC 700928 and S.epidermidis ATCC 35983,prior to affinity selection of clones binding to S.aureus to ensure specificty. 2013 HSACDLPMHPMC(6)[0.688 ± 0.033] HSACDLLMHPMC(1)[0.659 ± 0.033] HSACDLPKAPWC(1)[0.702 ± 0.035] 3 Phage display (common panning) 6 PMID:22945508 CD59 T-cell surface antigen CD2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA In phage ELISA, the absorbance at 490 nm was determined. Data were reproduced from the graph and expressed as means ± SDs. The absorbances of phages with the same sequence were calculated as mean ± SD. The absorbance of the wild-type M13 phage was 0.121 ± 0.006. The affinities of the 13 phage clones to human CD59 were measured by ELISA.Eight clones showed a high binding to CD59.All sequences exhibited 9 hydrophobic residues. 2014 GPGTWRA GPGTWRT GPGTYRA GHGTFRA GEGTWRA GPLTWRA GPSTWRA GPNTWRS GPYTWRS 9 Phage display (common panning) 3 PMID:22869370 cAb29 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2015 LLADTTHHRPWT APLQPRSDNPFR SATTPLIFPQTT YPAPQPLVTKTS ERSWTLDSALSM QHSAAHYSTRLS VDAQSKSYTLHD SFHQLPARSPLP 8 Phage display (common panning) 5 PMID:22941279 Boron nitride nanospheres,NNS Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Of these clones, LLADTTHHRPWT had the highest frequency .The BNNS-binding peptides were significantly rich in aromatic residues, including histidine (H) and tryptophan (W). 2016 CHSNYNPRC CHYHEQLRC CKSMEADTC CQKQPHSVC CQKQDSTWC CQTMLSLNC CPAQLLNAC CTVDKPPGC CQPPFPRPC CRFNTPLPC CRHSANQTC CYQDPRQPC CEGLSEIEC CPQLSDAGC CLPLRSIQC CLPNIDAKC CPNSRVAAC CRPSHATPC CSAGFTWLC CSLEHTWKC CRSPHTTYC CIESPTYWC CHQSSPKLC CQSSPPSLC CLITPSRLC CNPDRYYTC CNSTHKMFC 27 Phage display (subtractive panning) 1 PMID:22661481 Mouse retina Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) RA retinas was used in the negative selection.The OIR retinas was used in positive selection. 2017 CSTEALRHC(5) CSHEALRHC(2) CSSEAYRHC(1) CNMDTARLC(1) CSTTPSRMC(1) CDSTTPRSC(1) CSQPTLQAC(1) CTPTTSPAC(1) CMHSGPMAC(1) CANLHHQQC(1) CSEYLWSTC(1) CNFTWTSLC(1) CNKLEPFRC(1) CYPTHPYSC(1) CKWSPPQSC(1) CMSPKNVSC(1) CPTAKSASC(1) CLAPTANSC(1) CNQKTTSSC(1) CVDKSFNIC(1) CGLNVMRYC(1) CPDNKLRQC(1) 22 Phage display (subtractive panning) 2 PMID:22661481 Mouse retina Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) RA retinas was used in the negative selection.The OIR retinas was used in positive selection. 2018 CSTEALRHC(15) CDTTNSHLC(4) CNQKTTSSC(2) CNSYTNAAC(1) CQQPTSGHC(1) CTERQTRFC(1) CRDMGNHIC(1) CKTPFQHTC(1) CGLNVMRYC(1) 9 Phage display (subtractive panning) 3 PMID:22661481 Mouse retina Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) RA retinas was used in the negative selection.The OIR retinas was used in positive selection. 2019 CSTEALRHC(15) CLQMTKNSC(2) CNQKTTSSC(1) CTERQTRFC(1) CANLHHQQC(1) CSEYLWSTC(1) 6 Phage display (subtractive panning) 4 PMID:22661481 Mouse retina Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) RA retinas was used in the negative selection.The OIR retinas was used in positive selection. 2020 CSTEALRHC(23) CNQKTTSSC(1) CTERQTRFC(1) CSEYLWSTC(1) CKTPFQHTC(1) 5 Phage display (subtractive panning) 5 PMID:22661481 Mouse retina Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) RA retinas was used in the negative selection.The OIR retinas was used in positive selection. The phage harboring a peptide insert consisting of STEALRH distinguished between abnormal neovessels and normal vasculature. 2021 CSGGGPGVC CRLGGGLAC CWWGGGVSC CGSARGGGC CARGGGIRC CRAAGGGGC CGSSAGGGC CLGEAGGGC CGGLEGGGC CGNGGGESC CSTGGGCSC CLGGGEEWC G-G-G 12 Phage display (in vivo) PMID:11821895 Human bone Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2022 CHGFSHHGC CRRGFSLGC CGGFSPWLC CGRLVGFSC CTTGVGFSC G-F-S 5 Phage display (in vivo) PMID:11821895 Human marrow Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2023 CAEEGGTSC CEGGSFNWC CIEGGQVGC CEGGSVESC CEGGIFWHC CEGGLSGCC CCAEGGASC CAEGGVRGC CAEGGRVYC CVVEGGVKC CVLVGEGGC CTKKLEGGC E-G-G 12 Phage display (in vivo) PMID:11821895 Human fat Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2024 CGGLSPNWC CTGHLSPGC CVLSPGLGC CLSPGVKGC CLSPWKKRC CAWLSPARC CAWRRLSPC CLSPDDALC L-S-P 8 Phage display (in vivo) PMID:11821895 Human fat Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2025 CLVSGGMAC CLVSGCNTC CDLVSGYGC CLVSTSATC CTALVSQTC CWLVSGIGC CLVSSVFPC CPSLVSSVC CGVSLVSTC CQLVSGEPC CNLVSRRLC CLVSWRGSC CDHFLVSPC CGRGLVSLC CFPVALVSC CRWSSLVSC CWSKSLVSC CPGRSLVSC L-V-S 18 Phage display (in vivo) PMID:11821895 Human skeletal muscle Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2026 CGRRAGGSC CTRRAGGGC CSRAGGLGC CSYAGGLGC CDVAGGLGC CGAGGLGAC CGAGGWGVC CAGGTFKPC CLGEVAGGC CGSNDAGGC CYRGIAGGC CAGGVAGGC CGGLAGGFC CLLAGGVLC CLVVSAGGC CRTQAGGVC CAGGFGEQC CAGGLIDVC CAGGSTWTC CAGGDWWWC CAGGGLLMC CVAAGGGLC CLYGAGGSC CCALAGGCC CIGAGGVHC A-G-G 25 Phage display (in vivo) PMID:11821895 Human prostate Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2027 CGRRGSAGC(2) CRPGRRGSC(1) CSGRRGPRC(1) CGLGRRNGC(1) CGGRRSQTC(1) CLWDGRRHC(1) CGRRSVLTC(1) CFGRRNLFC(1) CGAGRRYWC(1) CGRRLWATC(1) CGVGRRFGC(1) CLEMVGRRC(1) CLSSIGRRC(1) CGRRWIDVC(1) CGRREEGLC(1) CGRRVLGRC(1) CRGLMGRRC(1) CRFLLGRRC(1) CPGVGRRLC(1) CGVIDGRRC(1) CADGRRLGC(1) CAGRRAQIC(1) CYGRRAREC(1) CPGRRLRMC(1) CGGRRVTLC(1) CEQGGRRLC(1) CSGRRLHPC(1) CFDHSGRRC(1) CGRRDVAIC(1) G-R-R 29 Phage display (in vivo) PMID:11821895 Human skin Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2028 CGGHPRLAC CGGHWRVNC CGGHILEVC CGGHRAQSC CGDGGHRPC CSCVGGHSC CGSGVGGHC CVRGWGGHC CWRGWGGHC G-G-H 9 Phage display (in vivo) PMID:11821895 Human skin Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2029 CWGSKGTVC CTGSLGTVC CWGTVSDAC CATGTVGPC CVVGTVAWC CWVVGTVTC CRVVHGTVC CGTVRFFSC G-T-V 8 Phage display (in vivo) PMID:11821895 Human skin Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2030 CPKHGVLWC CSGVLWYHC CGVLWAFGC CQARGVLWC CGVLVSRMC CGTVGVLVC CVGVLLPAC CGGVLLLSC CSGVLIHDC CPYFGVLAC CFFVSGVLC CLLAGGVLC CGEMGGVLC CGRAYGVLC CSGVLDG CWWGGVLGC CVWSRGVLC CGVLRGVSC CSFGVLRGC CKGSVGVLC CGGHFGVLC CWMDVGVLC CAFRVGVLC CGVGVLRKC G-V-L 35 Phage display (in vivo) PMID:11821895 Human multiple organs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2031 CMEGRGAGC CSEGRGFMC CVEGRNSKC CVEGRYTPC CFNEGRQMC CFEGRSRSC CDHVVEGRC CWDGTEGRC CLDWREGRC CRGCEGRVC CMTPEGRVC CRLFEGRVC CREGRRMCC CTQFEGRRC CSMEGRMFC CPGSAEGRC CGEGRILAC CEGRFSAWC CEGRSDIWC CEGRARWLC CEGRERWRC E-G-R 21 Phage display (in vivo) PMID:11821895 Human multiple organs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2032 CCQCGFGVC(1) CRGGFGVRC(1) CAVGFGVIC(2) CIVGFGVAC(1) CGNFGVVWC(1) CDEPFGVAC(1) CVWFGVGSC(1) CWFGVSLSC(1) CFGVGQWAC(1) CSMRFGVSC(1) CRFGVWTGC(1) CRFGVGRVC(1) CSGLFGVYC(1) CMKGVFGVC(1) CAFGVVSDC(1) CLYAFGVVC(1) CKVFGVVEC(1) CFGVRTDLC(1) CTIFGVRRC(1) F-G-V 19 Phage display (in vivo) PMID:11821895 Human multiple organs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2033 CVWPRFGGC(1) CSRFGGRVC(1) CMKFGGRLC(1) CRFGGALRC(1) CERFGGDEC(1) CFGGSVAPC(1) CWFGGSVQC(1) CFGGSWSLC(1) CLLFGGSAC(1) CMRLFGGTC(1) CFGGFFMYC(2) CEFGGQMNC(1) CTFGGLILC(1) CGNSFGGWC(1) CRTFGGAGC(1) CWVFGGKSC(1) CRGFGGLSC(1) CLWPSFGGC(1) F-G-G 18 Phage display (in vivo) PMID:11821895 Human multiple organs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2034 CGERISGPC CGERLSSRC CTEGERAGC CWWLGERVC CWAWAGERC CGVISGERC CGPGGERGC CLGGGERDC CDIAGERVC CSRSKGERC CKRKGERVC CSRPGERQC CCMRRGERC CTLRGERNC CFGERNRIC CRGERWDLC CGERTALLC G-E-R 17 Phage display (in vivo) PMID:11821895 Human multiple organs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2035 CPSGTSSWC CSMSGTGMC CLFDVSGTC CVTGLSGTC CNMVISGTC CGVSGTLGC CRSGTPGKC CGRSGTSGC CIYSGTLWC CCSGTLFCC CRSGTLQTC CLGSGTWSC CESGTATGC CFTERSGTC CRYLRSGTC CPLGSSGTC S-G-T 16 Phage display (in vivo) PMID:11821895 Human multiple organs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A patient received an intravenous infusion of the unselected random phage library, and 15 min after infusion tissue biopsies were obtained to provide histopathological diagnosis and to recover phage from various organs. 2036 CTARSPWIC(12) 1 Phage display (common panning) 3 PMID:22797828 Nd2O3 nanocrystals Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) This phage displayed sequence TARSPWI flanked by two cysteines exhibited about 24,000 times greater affinity towards Nd2O3 nanocrystals than a random phage in the same-tube competitive binding assay. 2037 AYQRFDDVASRF(10) VNLRMDDHDWSR(6) KWNMNEDQIFFR(3) AYKQTYHETTWF(3) INLQTSTLMSHT(1) IHLDHGTVAPIW(1) [ED]-D 6 Phage display (competitive panning) 5 PMID:22868630 Human mammary carcinoma cell line SK-BR-3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After the first selection, the acid eluted buffer was changed with Herceptin?. The shown sequences of peptides revealed that 40% peptides shared a consensus sequence (AYQRFDDVASRF).Consensus amino acid motifs (DD/ED) appeared at high frequencies. 2038 AEHYTFPKHSVWDIPYI AEQTPKHSLWTPYTYPA AEAHYKHSCRQDFCPTL K-H-S 3 Phage display (common panning) 4 DOI:10.1002/ddr.430330203 Human Platele Not determined. Not determined. Not determined. X15 M13 phage display library 2039 AEKNIVTYIPRGDMPTM AEYKHPRGDSPTHHQFG AEARIMRGDMPSSNHFT 3 Phage display (common panning) 4 DOI:10.1002/ddr.430330203 Human Platele Not determined. Not determined. Not determined. X15 M13 phage display library 2040 AEPGLAISCKRDVCITS 1 Phage display (common panning) 4 DOI:10.1002/ddr.430330203 Human Platele Not determined. Not determined. Not determined. X15 M13 phage display library 2041 AETQYPHPVCRGDCNMW AEPRGDWSTFRTHDTLF R-G-D 2 Phage display (common panning) 4 DOI:10.1002/ddr.430330203 Human Platele Not determined. Not determined. Not determined. X15 M13 phage display library Peptides contained KGD sequences suggesting that these fusion peptides bound to the glycoprotein IIb-IIIa(GPIIb-llla). 2042 AETLLDQGGHAVQLGDV 1 Phage display (common panning) 4 DOI:10.1002/ddr.430330203 Human Platele Not determined. Not determined. Not determined. X15 M13 phage display library 2043 AEFSKKTSEAFPVWRLM AEASVRSYLHKRLEHPL AEFYRPSLLTAWYSNMP AESASFSCRSDYCITST AELFSTHYLAFKEDYSQ AEEKHWLYVPDWTTAP AEEKHWLYSPCLVHCP 7 Phage display (common panning) 4 DOI:10.1002/ddr.430330203 Human Platele Not determined. Not determined. Not determined. X15 M13 phage display library 2044 YRHSVI LRHSVI WRHSVV LRHSVV R-H-S-V 4 Phage display (common panning) 3 PMID:1376364 Anti-P53 monoclonal antibody PAb240 p53 protein Not determined. Not determined. f3-6mer phage display library (X6) ELISA A wild-type phage control gave a background signal of 0.080. Phage clones giving a signal above 0.230 (~65%) were considered positive (data not shown). The average signal obtained from the positive clones was 0.452; the maximal signal 0.757. All four sharing a common RHSV tetrapeptide core sequence.The C-terminal residue of the encoded hexapeptide as either valinr or isoleucine. 2045 ADGGAQGTA(21) PGPSRAHFL(19) LSSREPQAR(11) RLTRELYAQ(9) YTQKHKHQA(4) 5 Phage display (subtractive panning) 3 PMID:7504991 Sera from RA patients Not determined. Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA,Competition experiment The binding of the sera from RA patients and normal individual (both not included in the pool) to the phage displaying pepl (ADGGAQGTA) was investigated by ELISA. The percentage of the sera positive for pepl was significantly raised in RA (44%) compared to the controls (13%). Similar results were obtained with the other selected phages. Then the reactivity of the phage displaying pepl was investigated in detail. In competitive ELISA, the binding of the sera to phage-coated plates was significantly inhibited by the phage itself, whereas no inhibition was obtained with an unrelevant phage displaying a different peptide. In competitive inhibition a synthetic peptide corresponding to the peptide displayed by the phage (ADGGAQGTA), inhibited the binding of the sera to the phage by 56%. Phages isolated during the second round of panning on RA sera were incubated with 100 pg of ammonium-precipitated immunoglobulins obtained from a pool of 40 normal individuals. After panning for 10 min on a plate coated with streptavidin, the non adherent phages (depletion step) were collected. RA sera was used in the positive selection. Ammonium-precipitated immunoglobulins from a pool of 40 normal individuals were used in the negtive selection. 2046 APWLYGPA 1 Phage display (common panning) 3 PMID:7685300 Anti-LeY antigen monoclonal antibody B3 Not determined. Not determined. Not determined. X8 and CX8C phage display library pool Phage that bind to the biotinylated mAb B3 were enriched by several cycles of biopanning on streptavidin-coated petri dishes. Alanine-scanning mutagenesis of the sequence coding for this peptide indicates that four residues,PWLY,were critical for binding to the mAb. 2047 HGRFILPWWYAFSPS RFRGLISLSQVYLSP ARVSFWRYSSFAPTY W-Y-A-[WF]-S-P 3 Phage display (common panning) 3 PMID:9237904 Thomsen-Friedenreich antigen (T antigen, TF antigen) Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Wells of the microtiter plates were first coated with streptavidin, washed with TPBS (phosphate buffered saline, pH 7.4, 0.5% (v/v) Tween-20), and blocked with 3% (w/v) BSA prior to addition of biotinylated antigens. Phages were incubated with antigen prior to washing and elution. In the last two rounds of some procedures, phages were pre-incubated with the biotinylated antigens before the streptavidin capture. A putative consensus sequence of W-Y-A-[WF]-S-P was present in the most frequently occurring clone, P30(HGRFILPWWYAFSPS). 2048 GSWYAWSPLVPSAQI W-Y-A-[WF]-S-P 1 Phage display (common panning) 3 PMID:9237904 Thomsen-Friedenreich antigen (T antigen, TF antigen) Not determined. Not determined. Not determined. f88-15mer phage display library (X15) Wells of the microtiter plates were first coated with streptavidin, washed with TPBS (phosphate buffered saline, pH 7.4, 0.5% (v/v) Tween-20), and blocked with 3% (w/v) BSA prior to addition of biotinylated antigens. Phages were incubated with antigen prior to washing and elution. In the last two rounds of some procedures, phages were pre-incubated with the biotinylated antigens before the streptavidin capture. A putative consensus sequence of W-Y-A-[WF]-S-P was present in the most frequently occurring clone, P10(GSWYAWSPLVPSAQI). 2049 CGFECVRQCPERC 1 Phage display (in vivo) 3 PMID:9664085 Mouse lung Not determined. Not determined. Not determined. CX3CX3CX3C phage display library Mice were first anesthetized with Avertin and then injected intravenously (tail vein) with phage peptide library.Organs were then weighed, homogenized, and the phage were rescued by infection with K91kan bacteria. 2050 CGFELETC CTLRDRNC CIGEVEVC 3 Phage display (in vivo) 3 PMID:9664085 Mouse lung Not determined. Not determined. Not determined. fdMED1-based CX6C M13 phage display library Mice were first anesthetized with Avertin and then injected intravenously (tail vein) with phage peptide library.Organs were then weighed, homogenized, and the phage were rescued by infection with K91kan bacteria. 2051 CVALCREACGEGC 1 Phage display (in vivo) 3 PMID:9664085 Mouse skin Not determined. Not determined. Not determined. CX3CX3CX3C phage display library Mice were first anesthetized with Avertin and then injected intravenously (tail vein) with phage peptide library.Organs were then weighed, homogenized, and the phage were rescued by infection with K91kan bacteria. 2052 SWCEPGWCR 1 Phage display (in vivo) 3 PMID:9664085 Mouse pancreas Not determined. Not determined. Not determined. X2CX4CX FUSE5-based phage display library Mice were first anesthetized with Avertin and then injected intravenously (tail vein) with phage peptide library.Organs were then weighed, homogenized, and the phage were rescued by infection with K91kan bacteria. 2053 YSGKWGW 1 Phage display (in vivo) 3 PMID:9664085 Mouse intestine Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Mice were first anesthetized with Avertin and then injected intravenously (tail vein) with phage peptide library.Organs were then weighed, homogenized, and the phage were rescued by infection with K91kan bacteria. 2054 YSGKWGW 1 Phage display (in vivo) 3 PMID:9664085 Mouse intestine Not determined. Not determined. Not determined. X7 FUSE5-based phage display library Mice were first anesthetized with Avertin and then injected intravenously (tail vein) with phage peptide library.Organs were then weighed, homogenized, and the phage were rescued by infection with K91kan bacteria. 2055 GLSGGRS 1 Phage display (in vivo) 3 PMID:9664085 Mouse uterus Not determined. Not determined. Not determined. X7 FUSE5-based phage display library Mice were first anesthetized with Avertin and then injected intravenously (tail vein) with phage peptide library.Organs were then weighed, homogenized, and the phage were rescued by infection with K91kan bacteria. 2056 LMLPRAD 1 Phage display (in vivo) 3 PMID:9664085 Mouse adrenal gland Not determined. Not determined. Not determined. X7 FUSE5-based phage display library Mice were first anesthetized with Avertin and then injected intravenously (tail vein) with phage peptide library.Organs were then weighed, homogenized, and the phage were rescued by infection with K91kan bacteria. 2057 CSCFRDVCC CRDVVSVIC R-D-V 2 Phage display (in vivo) 3 PMID:9664085 Mouse retina Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Mice were first anesthetized with Avertin and then injected intravenously (tail vein) with phage peptide library.Organs were then weighed, homogenized, and the phage were rescued by infection with K91kan bacteria. 2058 CVWLWEQC CVWLWENC 2 Phage display (common panning) 3 PMID:9834225 Anti-vWF monoclonal antibody 82D6A3 von Willebrand factor (vWF) 2ADF, Not determined. CX6C phage display library The two phage clones carried an almost identical sequence,CVWLWEQC and CVWLWENC, with a substitution of an N for a Q at position 6 of the hexapeptide. 2059 TAASGVRSMH(16) LTLRWVGLMS(10) TAASGVRSMH(8) LTLRWVGLMS(7) GGGTRAGMKY(2) TAASGVRSMH(1) LTLRWVGLMS(1) WGKIEDPLRA(1) AGQTLTASGD(1) DLLAVSWLRA(1) SAERGVVAMS(1) AIHSELMWVS(1) FWTERAGWAY(1) MVWSKGPLFL(1) AGTRMSWEVL(1) VSRSSRWGSI(1) DAHVLVPRTP(1) AQGIVLQLAL(1) LSPLLSPATA(1) 19 Phage display (common panning) 5 PMID:10383148 NG2 Not determined. Not determined. Not determined. X10 phage display library 2060 CLRSGRGC(26) CLRSGKGC(5) CLRSGHGC(3) CLRSGRGC(2) CLRNGKGC(1) CLRSGTGC(1) CLRSGLGC(1) C-L-R-S-G-x-G-C 7 Phage display (common panning) 3 PMID:10429241 Matrix metalloproteinase-9, MMP-9 Not determined. Not determined. Not determined. CX5C+CX6C+CX7C+CX9 phage display library pool Most of the clones selected and analyzed by sequencing consisted of a highly conserved CLRSGXGC motif. 2061 CRRHWGFEFC CTTHWGFTLC CSLHWGFWWC C-x(2)-H-W-G-F-x(2)-C 3 Phage display (common panning) 3 PMID:10429241 Matrix metalloproteinase-9, MMP-9 Not determined. Not determined. Not determined. CX5C+CX6C+CX7C+CX9 phage display library pool Each of these peptides contained a second cysteine and occurred as a cyclic motif C-x(2)-H-W-G-F-x(2)-C. 2062 CQAWCMYGFCGDC(17) CVGVCTYGFCIEC(3) CRPVCLWGFCITC(3) CDSVCRWGFCLFC(2) CSDICVWGFCVEC(2) CDSFCVYGFCYEC(1) CATFCRFGFCYSC(1) [FYW]-G-F 7 Phage display (common panning) 3 PMID:10429241 Matrix metalloproteinase-9, MMP-9 Not determined. Not determined. Not determined. CX3CX4CX2C phage display library Panning of this library on MMP-9 yielded YGF, WGF, and FGF motifs, which are similar to the HWGF consensus. 2063 CISMCIPDCISWC(9) CLALCTPDCWWYC(1) CVWLCGPDCWQYC(1) CFWLCAPMCSLVC(1) CFWQCAPMCSLVC(1) CVQMCMPACFHLC(1) CWVACTPACFLIC(1) CYNLCSPSCWEYC(1) CFSWCSPSCWSVC(1) CYIACLPGCVMLC(1) 10 Phage display (common panning) 3 PMID:10429241 Matrix metalloproteinase-9, MMP-9 Not determined. Not determined. Not determined. CX3CX3CX3C phage display library The sequences that bound to MMP-9 from this library did not show a clear consensus except that a proline residue was present at the central position of the peptide. 2064 SSLVDLILF(2)[2.4][55.1 ± 1.1] SWTLGERWW(2)[3.6][NT] SWTLGDSWW(2)[3.2][NT] SVPWDIVWG(1)[3.5][NT] SIEYWLDWL(1)[3.1][NT] SDWRWETLW(1)[3.0][NT] SFGVWDFIL(1)[2.7][NT] 7 Phage display (subtractive panning) 1 PMID:11017047 KIX domain of p300/CBP Not determined. Not determined. Not determined. SX8 phage display library (SX8) ELISA,Fluorescence polarization In phage ELISA, apparent affinity (μM) is defined as the affinity of the KIX domain binding peptide when expressed as a pIII fusion on M13 bacteriophage. Besides, fluorescence polarization was used to measure the actual peptide affinity (μM). NT denotes not tested. Phage specific for GST were removed by immunodepletion with 100 μg of agarose-bound GST after cycles 1 and 2 of affinity purification. 2065 TSPLNIHNGQKL 1 Phage display (subtractive panning) 5 PMID:11118031 HNSCC cell line MDA167Tu Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The isolated phages were subtracted using NHFs. Recovered phages were subjected to five rounds of MDA167Tu-selection successively. These selections were then followed by three rounds of NHF subtraction in succession. 2066 THALWHT[0.914 ± 0.058] 1 Phage display (common panning) 3 PMID:11165262 The human cell lines 16HBE14o- Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Either THALWHT- or IPACPSC-displaying phages were added to 16HBE14o- cells. Measurement of the absorbances at 405 nm were performed. Data were reproduced from the graph and expressed as means of three independent experiments ± standard deviations. The value of the IPACPSC-displaying phage was 0.163 ± 0.012. 2067 MAPYPLQ LTSPNSL MGSPLKW MQARGNW MGTMPLN MTLAPLG LQPPWHL MTIPLSP LTLPPQV MSLNNLP MAVPRPL LTRVPVG MATSPLR MTMPPLH MTPMRTT LASPLRP APPDHAG LAHPPQT MADPRYW MTSVPPQ SAPFSPA FASPYLT MSQAPSV LATPLAY MATHPLS LSRAPGW MSPLPWW MTPLWHA MTLPTIT MQTFPVS MSLTAPR MTVSRLG FSAVPPS LSRAPTA LAGTSIL LQIPPLL MSTPAFP FAKLPSG LSNMPRY 39 Phage display (subtractive panning) 5 PMID:11230317 LOX-1 Oxidized low-density lipoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) A clearing step on LZRS-b-gal-transfected hepG2 cells was performed before incubation and recovery from LZRS-LOX-1-transfected cells. 2068 LATPPKP MATPKPL x-A-T-P-x(3) 2 Phage display (subtractive panning) 5 PMID:11230317 LOX-1 Oxidized low-density lipoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) A clearing step on LZRS-b-gal-transfected hepG2 cells was performed before incubation and recovery from LZRS-LOX-1-transfected cells. 2069 LSKMPVA LSKTPVS L-S-K-x-P-V-x 2 Phage display (subtractive panning) 5 PMID:11230317 LOX-1 Oxidized low-density lipoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) A clearing step on LZRS-b-gal-transfected hepG2 cells was performed before incubation and recovery from LZRS-LOX-1-transfected cells. 2070 MSWPPRT MSRPTTT M-S-x-P-x(2)-T 2 Phage display (subtractive panning) 5 PMID:11230317 LOX-1 Oxidized low-density lipoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) A clearing step on LZRS-b-gal-transfected hepG2 cells was performed before incubation and recovery from LZRS-LOX-1-transfected cells. 2071 FQTPPQL FQPFPRL F-Q-x(2)-P-x-L 2 Phage display (subtractive panning) 5 PMID:11230317 LOX-1 Oxidized low-density lipoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) A clearing step on LZRS-b-gal-transfected hepG2 cells was performed before incubation and recovery from LZRS-LOX-1-transfected cells. 2072 MTAPPIQ MTARPIK M-T-A-x-P-I-x 2 Phage display (subtractive panning) 5 PMID:11230317 LOX-1 Oxidized low-density lipoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) A clearing step on LZRS-b-gal-transfected hepG2 cells was performed before incubation and recovery from LZRS-LOX-1-transfected cells. 2073 MQPWSHP MQPPPTT M-Q-P-x(4) 2 Phage display (subtractive panning) 5 PMID:11230317 LOX-1 Oxidized low-density lipoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) A clearing step on LZRS-b-gal-transfected hepG2 cells was performed before incubation and recovery from LZRS-LOX-1-transfected cells. 2074 LTRPPYT LTRPPYH LTRPLTV L-T-R-P-[PL]-[YT]-x 3 Phage display (subtractive panning) 5 PMID:11230317 LOX-1 Oxidized low-density lipoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) A clearing step on LZRS-b-gal-transfected hepG2 cells was performed before incubation and recovery from LZRS-LOX-1-transfected cells. 2075 FTTPPGV LTTPPKV x-T-T-P-P-x-V 2 Phage display (subtractive panning) 5 PMID:11230317 LOX-1 Oxidized low-density lipoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) A clearing step on LZRS-b-gal-transfected hepG2 cells was performed before incubation and recovery from LZRS-LOX-1-transfected cells. 2076 LSIPPKA LSRPPMP L-S-x-P-P-x(2) 2 Phage display (subtractive panning) 5 PMID:11230317 LOX-1 Oxidized low-density lipoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) A clearing step on LZRS-b-gal-transfected hepG2 cells was performed before incubation and recovery from LZRS-LOX-1-transfected cells. 2077 LTPATAI LTPARAT L-T-P-A-x-A-x 2 Phage display (subtractive panning) 5 PMID:11230317 LOX-1 Oxidized low-density lipoprotein Not determined. Not determined. Ph.D.-7 phage display library (X7) A clearing step on LZRS-b-gal-transfected hepG2 cells was performed before incubation and recovery from LZRS-LOX-1-transfected cells. 2078 VSNSNWPSFPSS(6) 1 Phage display (common panning) 8 PMID:11245619 PMCA398 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2079 MCPKHPLGC(3) LCPKHPLGC(2) x-C-P-K-H-P-L-G-C 2 Phage display (common panning) 4 PMID:11498772 U87-MG glioma cells Not determined. Not determined. Not determined. CMTI phage display library 2080 ECHKSPKEC PCTKPHMGC WCITYPEGC QCFKNDSGC SCVKFPEEC PCPTDHQEC VCATSGLKC 7 Phage display (common panning) 4 PMID:11498772 U87-MG glioma cells Not determined. Not determined. Not determined. CMTI phage display library 2081 RVVCEYVFGRGAVCS(8) GCCFLVGGLCYSTCA(2) 2 Phage display (competitive panning) 3 PMID:11522014 von Willebrand factor, vWF Type I collagen Not determined. Not determined. LX15 phage display library The bound phages were specifically eluted with fibrillar human collagen type I (100 ????g/ml). After amplification of the phages, two additional rounds of panning were performed. 2082 HTSTYWWLDGAP[1.602 ± 0.039] HQLPQYWWLSPG[1.094 ± 0.033] YWWL 2 Phage display (competitive panning) 3 PMID:11581662 [PA63]7 complex Protective antigen, PA Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA ELISA was done to assess the specificity of phages displaying different peptides, or the unselected PhD12 library as a negative control. Absorbance at 450 nm was determined, reproduced from the graph and shown. The absorbance of the negative control was 0.200 ± 0.018. Phage encoding peptides were eluted in the each round of biopanning using PBS and intact PA. The P1(HTSTYWWLDGAP) and P2(HQLPQYWWLSPG) peptides share the hydrophobic sequence YWWL; this commonality suggests that this tetrapeptide may play a role in binding to PA63. 2083 CGNKRTRGC 1 Phage display (in vivo) 4 PMID:12053175 Mice carrying human MDA-MB-435 breast carcinoma xenografts Not determined. Not determined. Not determined. CX7C T7 phage display library For the ex vivo screening, MDA-MB-435 human breast carcinoma xenograft tumors cell suspension was incubated with the phage library. The phage selection process was repeated 3 times. Then the ex vivo pre-selected phage pool was then subjected to an in vivo selection round by injecting it into the tail vein of a nude mouse bearing an MDA-MB-435 tumor. 2084 QIDRWFDAVQWL(24) 1 Phage display (subtractive panning) 5 PMID:12082461 Human melanoma cell line MeWo Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Three subtractions with HeLa cells before the first selection on WiDr cells. No further subtractions were done before the four next selections. 2085 HEWSYLAPYPWF(13) 1 Phage display (subtractive panning) PMID:12082461 Human melanoma cell line MeWo Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Three subtractions with HeLa cells before the first selection on WiDr cells, and three subtractions with 293 cells before each of the subsequent four selections on the WiDr cells. Twenty-four phages were isolated from the library and thirteen of them harbored the same peptide sequence (HEWSYLAPYPWF). 2086 YQATPARFYTNT(4) 1 Phage display (subtractive panning) PMID:12082461 Human breast cancer cell line MDA-MB-231 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Three subtractions with HeLa cells before the first selection on MDA-MB-231 cells, and three subtractions with 293 cells before each of the subsequent four selections on the MDA-MB-231 cells. Six phages were isolated from the library and four of them habored the same peptide sequence YQATPARFYTNT. 2087 CGWMGLELC(11) 1 Phage display (subtractive panning) 5 PMID:12082461 Human breast cancer cell line MDA-MB-231 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Three subtractions with 293 before each of the four subsequent selections on MDA-MB-231 cells. Fourteen phages were isolated from the library and eleven of them possessed the petide sequence CGWMGLELC. 2088 CISPDAHSC(9) CTLSHTRTC(1) 2 Phage display (common panning) 5 PMID:23128437 Anti-CK 8/18 complex monoclonal antibody Cytokeratin (CK) 8/18 complex Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The reactivity of selected phages against K94 antibody was examined by phage ELISA. Empty phage (Eph) without an insert peptide sequence and XC20p1 phage were used as control antigens and other HCC-derived auto antibodies (XC20 and XC90) were used as control antibodies. Results showed that K94p1 mimotope phage (displaying CISPDAHSC) showed high reactivity to K94. Other phages expressing peptide sequences of CTLSHTRTC (K94p7), CLSIGMPGC (XC20p1), or phage without the insert peptide sequence (Eph) showed no binding to K94. 2089 SLHYSRDQLVAL 1 Phage display (common panning) 3 PMID:23094039 Eukaryotic initiation factor (eIF)4E Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Biotinylated full length eIF4E was loaded onto 10 ml of steptavdin M280 magnetic Dynabeads (Invitrogen). Each selection was performed on the beads. 2090 TMGANLGLESPE(1) TMGANLGLKWPV(5) TTGANLGPKWPV(1) TMGANLGLKWPV 3 Phage display (common panning) 3 PMID:23193370 Zinc oxide, ZnO Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2091 FWLPSPT WWRPPVL WWKYPPQ YWFPAPP SWFPLRS YWLPYFS WWLPPGS W-x-P 7 Phage display (common panning) 4 PMID:23119051 Karilysin catalytic domain, Kly18 Not determined. Not determined. Not determined. Ph.D.-7 and Ph.D.-C7C phage display library pool 2092 HRHRRRE HKHRRRD HRHRRRE HKHRRRD H-[RK]-H-R-R-R-[ED] 4 Phage display (common panning) 3 PMID:23033479 Calretinin, CR Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) A consensus CR-binding sequence, H(R/K)HRRR(E/D), consisting of five or six basic residues (His/Arg) flanked by an acidic (Glu/Asp) residue was identified. 2093 STFCLLGQKDQSYCFTI(2) HMRCLHYKGRRVCFLL(5) KTMCLRYNHDKVCFRI(2) LVLCLVHRTSKHRKCFVI(1) 4 Phage display (common panning) 5 PMID:23076147 Human IgA, hIgA Not determined. Not determined. Not determined. X3CX7-10CX3 T7 phage display library 2094 FVSRSNSLLGLLLDGAINP VNPVSSRLIALLYDDVNDG HRLLVNSLDVLLSVDGPNL SFTSTGLLNILLSATNADS FRNGCFNLYRLLDSNFAAN PYRHRALLGFLLNDFSDHH CRYHSIILSHLLSSGDYSH RNFSRNILYNLLHCPLNDN DIFSSSNLSRLLSSPYYPG RDSRHCSLSSLLSGGCSVI FASGHSTLRDLLCNGGSYS SSCGTSRLCSLLNYSVVYS ADFPSATLNRLLLCYSNNL VGRSHFALGTLLSRDFALC VRRYSFHLAALLSSASGGH GDGADRFLFNLLVSGGSIH ACDISHSLRYLLLHSCGSS LPPASSLLFNLLHSSNINY FGIGTSALGHLLSASGCRS GSYNSPNLRCLLVGRDSCR CSDGGSILTLLLCSNVSRA GTDGRCTLRCLLTRPYSNP PCRSSSVLYHLLSSRGYGF SAISGSFLYNLLSSRVYSN GRVSGSRLRDLLRSYPAHT VLGVHHSLSNLLRGSLYTS AVGNNSTLFNLLRNTNHYV INRPGTDLARLLRYSISVS VVVYTNILHNLLSRNNTNF RTSNVSRLFSLLSRSSLAF LDRVRGILFNLLGTPFRSV L-x-x-L-L 31 Phage display (common panning) 3-4 PMID:22930062 Unphosphorylated ERα LBD Estrogen Not determined. Not determined. X7-LXXLL-X7 T7 phage display library 2095 DHHTNGSLYFLLSNNGDNN HRLLVNSLDVLLSVDGPNL FSVAIGNLGTLLVLDLDRN AFNRPSFLFNLLTSDTHSL PYRHRALLGFLLNDFSDHH FSVNSFRLYHLLATSHGTD APSPHVNLISLLTSSSSSG PIHGSSILHNLLSGNFTAF FASGHSTLRDLLCNGGSYS SSCGTSRLCSLLNYSVVYS TDFHFCTLSPLLDVTRYSY IGCRFHYLSGLLPFGGPTF VYCVGSRLYYLLSSNCHPS VRRYSFHLAALLSSASGGH RDARPSALYRLLSRHHDGC SYTSHSTLACLLIGRDVHN YTVHFGDLRCLLDRPFCHS CSDGGSILTLLLCSNVSRA GTDGRCTLRCLLTRPYSNP PCRSSSVLYHLLSSRGYGF AVTAHSRLYALLGVGYPYR GRVSGSRLRDLLRSYPAHT FHDPHGTLFGLLSGRSSFG NIPANSVLFNLLSAGVNHR GSYNNGALFDLLIPSTRST GSYNNGALFDLLIPSTRST DTVCFSRLRRLLISSHHNR 27 Phage display (common panning) 3-4 PMID:22930062 Phosphorylated ERα LBD, ERα LBD-P Estrogen Not determined. Not determined. X7-LXXLL-X7 T7 phage display library 2096 AISRGRVLISLLHDHYDSL(1) CSDGGSILLLLLCSNVSRA(1) CSDGGSILTLLLCSNVSRA(1) DCVGARTLVALLNDCDAVV(1) DDGSHIALHPLLFGHLVPF(1) DNVTGIFLHSLLLGNDPVG(1) FASGHSTLRDLLCNGGSYS(2) FPASYTTLRSLLVGYADSF(1) FVSASSVLFTLLNARSSGV(1) GRVSGSRLRDLLRSYPAHT(1) GTDGRCTLRCLLTRPYSNP(2) IIDDHGSLLSLLPLIGTSF(1) INRPGTDLARLLRYSISVS(1) LDDHHGLLTHLLGDYSYSF(1) LTGRRNVLTALLFDGSLVG(1) NAFLRGFLADLLAFPSNHR(1) NDYSHGRLRHLLNDFCTGF(1) PCRSSSVLYHLLSSRGYGF(1) PYRHRALLGFLLNDFSDHH(1) RAVCFRSLAYLLTDGRSVN(1) RAVCFRSLXYLLTDGRSVN(1) SGRIGSALICLLSGPTDVS(1) SHDNRNDLCYLLRSTNVCF(1) SNHPGVNLRYLLHNVHSNF(1) SRIHGNFLFHLLSGNVGSN(1) SSCGTSRLCSLLNYSVVYS(1) VALHGNNLSFLLGHSTGHS(1) VFARVPSLDILLDFGCGLP(1) VNPVSSRLIALLYDDVNDG(1) VRRYSFHLAALLSSASGGH(1) VSSTNNTLRHLLSAAGYYG(1) VSTARLRLSSLLGNSYGLG(1) YFSYSSILDRLLSIDGDYH(1) YGRLSYNLVSLLGSVGSHN(1) YLHTRHFLISLLSDDGPNN(1) YPDTHLLLYSLLLPDSANA(1) 38 Phage display (common panning) 3-4 PMID:22930062 Unphosphorylated ERβ LBD Estrogen Not determined. Not determined. X7-LXXLL-X7 T7 phage display library 2097 VNPVSSRLIALLYDDVNDG YSSGSCILHCLLDSNLCDC HTCGSSILLGLLNVDCSYD VPLFDCYLSVLLDPGLPVN CSPFLINLSRLLLIPDFNL SDGFGTLLIGLLYDFVGNP RGAVGSSLVCLLTYSDNVC ALNSNVGLVRLLLRDDFSS PYRHRALLGFLLNDFSDHH TYNYHPLLVSLLGANFDGH PIHGSSILHNLLSGNFTAF RVPNLGILGRLLFGGATIP APGRVFSLGRLLNSSSFVS FASGHSTLRDLLCNGGSYS YLSNYHNLIRLLGRASYDP YRPCHLSLISLLNHDSHLN RNHISPGLFYLLPYRNIID VRRYSFHLAALLSSASGGH RLHDISFLEYLLSGSGHVV CFALVSRLFCLLVCTISHA CGHVDGLLSALLGYPVPCR CSDGGSILTLLLCSNVSRA GYAYRSNLNFLLGSNCRNF SSFCFGSLYSLLATHRCSS GTDGRCTLRCLLTRPYSNP PCRSSSVLYHLLSSRGYGF SAISGSFLYNLLSSRVYSN ICSNSSRLSDLLVSSSYLR GGNRFSPLYRLLVRYGSNS GRVSGSRLRDLLRSYPAHT FDVTSSGLASLLTRVFSFS SDFNRAILNRLLVIVRSGT FVSASSVLFTLLNARSSGV NAFLRGFLADLLAFPSNHR 34 Phage display (common panning) 3-4 PMID:22930062 Phosphorylated ERβ LBD, ERβ LBD-P Estrogen Not determined. Not determined. X7-LXXLL-X7 T7 phage display library 2098 SWFHPQRRHSHQ SWMPHPRWSPQH MHRAPSTHKLLP ASWHQHYMKHKP SEFHRHGDKEHK CEFPRSWDMETN SLTRHKPEPHRK SLTWHKHELHRK SPPWHKHELHRK GGWHKHISRSDP YHKHPHTYHNFK HPKHPHTHTNDQ HMHMHQHVAWTQ HMGMTKINYSAL SNYSDVKRLPTV SVNWQKQTISNL 16 Phage display (common panning) 3 PMID:23053512 Anti-PI-LPS monoclonal antibody 1E4 Phase I LPS, PI-LPS Not determined. Not determined. Ph.D.-12 phage display library (X12) 2099 ETYAGNG(3) QLPNRTV(3) HLAALEY(2) AATVISR(2) VTSPLLI(2) QPRSPSA(2) TQPRYPL(2) SDTLPKM(2) LPRLAMN(2) TTWNPLD(2) RAPFVSY(2) GKPAHPT(2) ASSLLNG(1) HPKWHPS(1) KSPLAPL(1) VNLGTRQ(1) SSHSNLQ(1) TSFSVLA(1) NFTSRPT(1) NYTSQLT(1) VMATIAP(1) YATHLPA(1) YATDPHK(1) YIHAPAP(1) LVTSPSP(1) VLRPNLP(1) DPTLPLN(1) HPSLPKL(1) NTRLPMP(1) HNTTWSS(1) RVPTWPS(1) VSEVARV(1) MSENPRT(1) APLVRLP(1) KGALPIY(1) HIPYDPP(1) NPPSLWA(1) YSGKHSS(1) HMSKAFH(1) 39 Phage display (common panning) PMID:23104667 Bone marrow-derived dendritic cells, BMDCs Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2100 CQPAPGKQC(4) CNPASSQLC(3) CSKSSDYQC(2) CTQKSGPVC(2) CTADQQRQC(2) CTHSSLHGC(2) CNVQNTRNC(2) CSQLSAHSC(2) CLTNFHRTC(2) CTPASARSC(2) CQLDHLKAC(2) CTKYQATMC(2) CYEHGKPLC(2) CHGKPSANC(2) CSTMRSPTC(1) CSTKNYREC(1) CHERQVHAC(1) CSNSNPKSC(1) CLPSAQMTC(1) CPKNGSDPC(1) CNDAAWQQC(1) CTLPLSPAC(1) CQPHKAKSC(1) CIPNAAARC(1) CLGQPYHTC(1) CVPGHRGTC(1) CQSPKNTTC(1) CPLQDPLRC(1) CNHGSFMHC(1) CGNSAHRSC(1) CLEATQKHC(1) CTTPGNLQC(1) CNPTATPLC(1) CITQKQAAC(1) CTSTVRPSC(1) CPGLHTTQC(1) CKTPGPATC(1) CSHESHRQC(1) CTSRPMAAC(1) CVGHKPQSC(1) CPANGAMHC(1) CNQGPVPSC(1) CSQTHGQPC(1) CPHQGSPLC(1) CHATAQATC(1) CWSVNQPFC(1) CLMTPTHTC(1) CLNSKLSHC(1) CTRLAQNTC(1) CSTSSTSLC(1) CPKSLKNTC(1) CHQAANSTC(1) CDTPITPAC(1) CQPMNSLTC(1) CSFSQNGEC(1) CTTTKIALC(1) CNQAILYKC(1) CSSSPSGGC(1) CQARLTAQC(1) CHPNTADYC(1) CPTNSINGC(1) CHSSPTRQC(1) CQAKSQGSC(1) CERWHNKHC(1) CTGASHQKC(1) CTMQNAMTC(1) CATPHHRVC(1) CDPKSMPKC(1) CNTQTTKIC(1) CAPSSKSLC(1) CSTKMLPRC(1) CKEHASVHC(1) CNDQARQIC(1) CMAGTPTNC(1) CDKGRTQLC(1) CRTPNHNTC(1) CQGPSLMAC(1) CTTSKSLMC(1) CTNSDTRHC(1) CSSAEAFSC(1) CHAPQLRHC(1) CSPHTPSLC(1) CPPTTMHAC(1) CDGLNSKNC(1) CATSYGLPC(1) CQPHHSPVC(1) CKSPVHGQC(1) CMNPSQSSC(1) CGLAVNSTC(1) CTGPHSFHC(1) CNLTNKALC(1) CPTGALDFC(1) CGTNSTPTC(1) CTWEQKKIC(1) CDLKHPASC(1) CKPSKLDIC(1) CDSHKDLKC(1) CPNNLASSC(1) CYAPGKSHC(1) CSYSPLHLC(1) CSLTSSRTC(1) CMNTMAAAC(1) CFSSTPTRC(1) CSYPMPKMC(1) CGYLAHSAC(1) CTPSPGEVC(1) CSTTSHRLC(1) CSEGYTSHC(1) CTSHSHPLC(1) CDMSDRSLC(1) CNGERTGQC(1) CKSTAPHLC(1) CPYQAQKPC(1) CSINRDRLC(1) CLPNNRASC(1) CPALQESKC(1) CMTTPTAHC(1) CDNPQFSSC(1) CELGPSRSC(1) CSEKWSKVC(1) CMGAKAPTC(1) CTNPATTQC(1) CDLSRTALC(1) CPTLKSSAC(1) CSHQGYNTC(1) CTSSQHTNC(1) CESPNSLKC(1) CPKPGATEC(1) CPKTPSNLC(1) CSSTAMGLC(1) CSPLSPRQC(1) CRQGSLPNC(1) CEMPGAMRC(1) CTDKAGPNC(1) CTNPKGVLC(1) CRVPSGSQC(1) CPTPKISAC(1) CSSSNPYFC(1) CSPRHANTC(1) CDATSAQLC(1) CKDHQTPTC(1) CVHMSRNYC(1) CHAYHKSNC(1) CLHHSLHMC(1) CPPQKGGRC(1) CQTNHTPSC(1) CNQTSLSAC(1) CGTLHKYPC(1) CTDMFRSLC(1) CSRTSDYQC(1) CMPNTKPLC(1) CDQDQGHTC(1) CDMKQGPSC(1) CDFRMQNQC(1) CTSTNSQHC(1) CPNNLASLC(1) CHNINTKQC(1) CMPDSTKQC(1) CNHSLPKNC(1) CPLSQPRPC(1) CPLSPSTLC(1) CTSGAERSC(1) CNKSLNPQC(1) CLVRQVAQC(1) CPLGKLQTC(1) CQANPLAMC(1) CPTAPNHSC(1) CSDPSRMRC(1) CSARHTAHC(1) CPLNDPSIC(1) CSPTTPKQC(1) CTPESKAMC(1) CVSASVRQC(1) CHPLLKAPC(1) CQVMSSKQC(1) CTQTKTFTC(1) CPFTFAPTC(1) CNASDNGLC(1) CNAESPMFC(1) CTSHQRPHC(1) CPTESYMLC(1) CIKTEGQHC(1) CLKSPRPDC(1) CTDTLRPAC(1) CYASTRLQC(1) CTAQAIYPC(1) CKASTPQAC(1) CHIPGPHRC(1) CETDSNAVC(1) CRNAPYPNC(1) 190 Phage display (in vivo) PMID:23104667 Mouse kidney Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The library prepared in 500 μl PBS was orally administered into starved male SD rats using an oral zonde, before they were lethally anesthetised after 1 h of retention and again perfused via the left heart ventricle with DMEM supplemented with 1% heparin to remove circulating blood from the organs. 2101 CQPAPGKQC(3) CSKSSDYQC(3) CYSRAAPQC(2) CPKVNSAIC(2) CTGPHSFHC(2) CSNERHAMC(2) CSSHKSTYC(2) CERWHNKHC(2) CSPSSNQMC(2) CLAQAMGSC(2) CPQNTAKAC(2) CDDGRRGHC(2) CPTPKISAC(2) CTENVHMLC(1) CNPASSQLC(1) CPSPLTSNC(1) CISSQPSHC(1) CNVRHQSAC(1) CSAKPALAC(1) CVASHNGHC(1) CDKNLQLHC(1) CDSLTYMHC(1) CRPNESLHC(1) CPLSGPALC(1) CSSSNPYFC(1) CTKYQATMC(1) CSLPLGADC(1) CTSISDKQC(1) CNPTRAASC(1) CMASHLDTC(1) CSSALIHQC(1) CKINERNLC(1) CHDWRLPHC(1) CPSALLNSC(1) CKPQFKPFC(1) CTGGIKATC(1) CVASPLLQC(1) CKPLSNWSC(1) CPRESPNLC(1) CPGTQHSLC(1) CSGTTPRYC(1) CDSTINRLC(1) CVLERHSSC(1) CPSTPTREC(1) CLHYFGPPC(1) CYAPGKSHC(1) CHNQDFHEC(1) CWGSHPAHC(1) CQSTNLKRC(1) CNGAPDHFC(1) CAPKMNQHC(1) CPGYSRATC(1) CTSKYNAQC(1) CKANTQELC(1) CTADYRSSC(1) CLQALPPVC(1) CQNKFPGAC(1) CMAHNGKSC(1) CQAKSQGSC(1) CNPGTLSAC(1) CLKISLQAC(1) CKQGQPHSC(1) CTYQQPKLC(1) CTPQTAGMC(1) CLPKPQQVC(1) CLQSPHQHC(1) CGTTIKNIC(1) CGVRTNLLC(1) CQEGLNHRC(1) CGNQPLAHC(1) CRTPPLLSC(1) CLRITSHQC(1) CENRSDKVC(1) CLSSTSNTC(1) CTQTKTFTC(1) CPASHKASC(1) CSQLSAHSC(1) CPDRFHVSC(1) CLPSQPSQC(1) CTQSFIRQC(1) CHKGPFQSC(1) CQPMNSLTC(1) CDPMSNRSC(1) CRIDKLLSC(1) CTHQFSRIC(1) CRAIHLDGC(1) CPNKDVAHC(1) CSITRPASC(1) CQGSLRTQC(1) CYQFQGRSC(1) CTAPQHRQC(1) CRYSTADQC(1) CQSSFHPTC(1) CKLPSKFHC(1) CHSLHSRLC(1) CQQFTYSSC(1) CNPASSQGC(1) CLPHWMQPC(1) CNSELNMQC(1) CTMTPRLWC(1) CDTRTLQSC(1) CEPRSPQQC(1) CNSMLTTQC(1) CPPHMKAFC(1) CTPAGHQHC(1) CHMHPYSRC(1) CKPQDSRTC(1) CLERSAASC(1) CTMTSGLPC(1) CHSAPSTEC(1) CHSSNPASC(1) CKLTGAQHC(1) CTQKSGPVC(1) CTSPSPLLC(1) CTSSQWPLC(1) CHHLRGPTC(1) CPATPDYSC(1) CNANARLQC(1) CPNEKEPDC(1) CEQVGGPRC(1) CMEAHSLSC(1) CPMFPNTSC(1) CVNRSDASC(1) CNHSTRLLC(1) CLFNNTNEC(1) CLPGAPHRC(1) CEQNLERAC(1) CDEHRPAIC(1) CNHGSFMHC(1) CTHSRLDHC(1) CTEKTTTVC(1) CPNPSTMKC(1) CHEEARPHC(1) CSQTATRMC(1) CPLSDKAHC(1) CHRHLTTSC(1) CHAYHKSNC(1) CNKTNMPTC(1) CSHVTQSLC(1) CGVLFTQDC(1) CSKHDFPGC(1) CSQIHPRAC(1) CFSPKSMTC(1) CNPKSAILC(1) CLDGSTPIC(1) CMEKTRPSC(1) CSSSERLPC(1) CSHSTYNTC(1) CNGRIPFQC(1) CQSHGAAHC(1) CNVSKASQC(1) CHRSMLGTC(1) CDSPTPMHC(1) CNQMRDHEC(1) CQRITDTLC(1) CGQMQLIRC(1) CQTINYGHC(1) CPSITRHHC(1) CPSPISQTC(1) CKDHESRQC(1) CLSLAAHFC(1) CNNNSNVEC(1) CDPQVNPAC(1) CDPNEMSQC(1) CSNKTEHAC(1) CMDDKAESC(1) CKNYNTNQC(1) CTAGSSLRC(1) CGVNPMSAC(1) CFGAHSPLC(1) CETQQPRTC(1) CTYLRGPNC(1) CHKGPFRSC(1) CVPASMRTC(1) CPEHTHQMC(1) CTADQQRQC(1) CPNNLASLC(1) CNDPQTRNC(1) CRTTTDQHC(1) CSVIGAGSC(1) CDATQMPHC(1) CHSLEIAFC(1) CTTTSHLSC(1) CPKILGSLC(1) CTPQNLRNC(1) CTWEQKKIC(1) CAVVEAQMC(1) CTDHTAPAC(1) CSTALSAGC(1) CPKNTHQPC(1) CDLASRHTC(1) CEPGQRLLC(1) CPLPMTHAC(1) CPQPSNASC(1) CGHDNNSAC(1) CPKSLKNTC(1) CSAPEILSC(1) CLGQHPDHC(1) CDPLSHNLC(1) CPASEFSRC(1) CPERLRHYC(1) CPLDKSQQC(1) CSTTGSTKC(1) 203 Phage display (in vivo) PMID:23104667 Mouse liver Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The library prepared in 500 μl PBS was orally administered into starved male SD rats using an oral zonde, before they were lethally anesthetised after 1 h of retention and again perfused via the left heart ventricle with DMEM supplemented with 1% heparin to remove circulating blood from the organs. 2102 CQPAPGKQC(4) CQPMNSLTC(3) CKPNDTHQC(3) CSSHKSTYC(3) CTADQQRQC(3) CTSHQRPHC(2) CSKSSDYQC(2) CPKSLKNTC(2) CTATRLHLC(2) CLERNYSAC(2) CRMPTIQYC(2) CTGPHSFHC(2) CNPPTAPQC(2) CTQKSGPVC(2) CYGNLLNQC(2) CNPRNTHWC(2) CTSTVRPSC(1) CVASLNWTC(1) CSPRHANTC(1) CTTPTKPEC(1) CSMPHLTSC(1) CLPYDKRIC(1) CSPAMEQTC(1) CALPSSQLC(1) CPVKPQSPC(1) CLQNTLPVC(1) CMKTTFPYC(1) CDLKNPTAC(1) CTTGVGFMC(1) CPRAVPALC(1) CKAEATRSC(1) CGHYANQSC(1) CPNNLASLC(1) CGSGPFPMC(1) CHKSAPALC(1) CMHANPKTC(1) CTTLLHRAC(1) CPHQRNVSC(1) CLSDLHPMC(1) CLRELNPQC(1) CSARHTAHC(1) CLHTRTPLC(1) CHQSDRASC(1) CKTEGQPFC(1) CPGTTATHC(1) CHPLNRNSC(1) CSTHTPEIC(1) CDGTSHPHC(1) CPSSLFPLC(1) CLQLLSPQC(1) CLPAFQPRC(1) CVSNGQPHC(1) CTLYNRPTC(1) CPKYNVPHC(1) CPLNSPHTC(1) CVNAKSVTC(1) CTHPGQLSC(1) CTPSSRGAC(1) CERWHNKHC(1) CPLLAPLHC(1) CMDRYSPRC(1) CFKNTHFSC(1) CDLESKALC(1) CPTLSHAAC(1) CQQTSQRMC(1) CNSSPWTAC(1) CPMTHLNMC(1) CAPATRPTC(1) CHNPGLQHC(1) CLMQQAANC(1) CSTALSAGC(1) CHTMSVAAC(1) CSILTHTRC(1) CERTSHHSC(1) CTSPHSPLC(1) CKPQSTMMC(1) CELNPSRSC(1) CLKHSSNLC(1) CPTPKISAC(1) CTPSQSSKC(1) CRVSDHIAC(1) CSLTKLSFC(1) CQDSKSQYC(1) CTSGPQRQC(1) CTDRPEKSC(1) CLPHWQSHC(1) CLDTNLTKC(1) CTSMSPTHC(1) CTAQNNHAC(1) CMALEGKVC(1) CHQHLLHKC(1) CLSPNKSHC(1) CTQLSEAQC(1) CYPSKLNNC(1) CDDGRRGHC(1) CTKHPFTHC(1) CPNTKHPLC(1) CLSSAWRAC(1) CLTPAQPQC(1) CPADRLETC(1) CNPFQRPSC(1) CSNTSNSVC(1) CLPPPFPRC(1) CPTAPLTQC(1) CLPGQSSIC(1) CLSTHPGRC(1) CSTPGPSQC(1) CSHKTPGLC(1) CHTGKLPYC(1) CSRATPEVC(1) CTSGIEKAC(1) CLYPWDTRC(1) CLHGFMDRC(1) CHKGPFQSC(1) CPATSHQVC(1) CSSDKTPVC(1) CSWRHQESC(1) CITKMSRSC(1) CSTPSGTTC(1) CGPTKHSEC(1) CNPASSQLC(1) CTQMGYTMC(1) CTTGMTPHC(1) CPSTSKGYC(1) CMAHNGKSC(1) CNAPHHKTC(1) CPTPYNLQC(1) CLENANRKC(1) CHWSRQDSC(1) CTTHVPSFC(1) CSSAKAIYC(1) CRPNESLHC(1) CPPNIRYQC(1) CTKSTVNAC(1) CVPVTRMVC(1) CPPTHTLMC(1) CTVNLAGTC(1) CKTTDFQQC(1) CSVPSTNRC(1) CYGLNTNAC(1) CDISGPLAC(1) CDRTHLAMC(1) CPKVNSAIC(1) CESTDRPIC(1) CSALARSNC(1) CDSTHPRSC(1) CWTTDPLHC(1) CDGRNLQLC(1) CTHESRTSC(1) CQPNDLQLC(1) CTQHIHTQC(1) CVSSHSTQC(1) CTADYRSSC(1) CPTLSNSYC(1) CHNSLKMVC(1) CLFNNTNEC(1) CHPSSRGIC(1) CGDRPSPSC(1) CQPGAHRLC(1) CTSTSYKMC(1) CNPPVAPQC(1) CKNSESPNC(1) CPNQFRQLC(1) CQTPILGRC(1) CTTVNLRNC(1) CQSTNLKRC(1) CIGLAKQLC(1) CNKMPGASC(1) CTKPDTYVC(1) CPNSIITHC(1) CSASPPKTC(1) CQGTPNPAC(1) CVHQKNPFC(1) CYAPGKSHC(1) CIMEQFTTC(1) CPLAQGSKC(1) CQTSKMPHC(1) CSTTSHRLC(1) CDHTHYSHC(1) CPLQSSSTC(1) CHTPKMPQC(1) CGSADRGRC(1) CTTLARSYC(1) CSSALIHQC(1) CPSPISQTC(1) CPNDRSWSC(1) CDTPYPHNC(1) CWPTTQKTC(1) CDQSHTPNC(1) CAPSSKSLC(1) CTPPLSLRC(1) CPGYAPAQC(1) CPAGAASRC(1) CDTNPLWSC(1) CSPRHPTMC(1) CPIPHVPHC(1) 196 Phage display (in vivo) PMID:23104667 Mouse lung Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The library prepared in 500 μl PBS was orally administered into starved male SD rats using an oral zonde, before they were lethally anesthetised after 1 h of retention and again perfused via the left heart ventricle with DMEM supplemented with 1% heparin to remove circulating blood from the organs. 2103 CQMKPHTTC(3) CSHKTPGLC(3) CTADQQRQC(3) CTGPHSFHC(2) CRTLPINVC(2) CHTGKLPYC(2) CHKGPFQSC(2) CYAPGKSHC(2) CSPHTPSLC(2) CSKSSDYQC(2) CNTNDVRRC(2) CMEKKMSNC(2) CDGNSKPFC(2) CSPSSNQMC(2) CTWEQKKIC(2) CQAPGESSC(2) CTDTTHPYC(2) CNPASSQLC(2) CQSTNLKRC(2) CPKSLKNTC(2) CSQLSAHSC(2) CPRESPNLC(1) CTTHQTRQC(1) CDPQVNPAC(1) CMDMIRTTC(1) CPKSPAIMC(1) CTTTPHNSC(1) CETGNRGHC(1) CPPSSPRHC(1) CYGTSRLTC(1) CTLPMSDRC(1) CPDHQRSTC(1) CPHPLSANC(1) CERWHNKHC(1) CPSPISQTC(1) CSDYPRTTC(1) CQWTMMREC(1) CNQPLLHTC(1) CHHSRQAQC(1) CLGQPYHTC(1) CTPASARSC(1) CSTDSQMLC(1) CQSRMSPTC(1) CTSTVRPSC(1) CEPTTAQWC(1) CGDTPHKAC(1) CPSSTSPRC(1) CRDPVLSTC(1) CNQTLQRSC(1) CEHNSALVC(1) CHPREPLSC(1) CNSNHNATC(1) CKTTQPLLC(1) CSAGQDTLC(1) CTPRSVGAC(1) CQTINLMTC(1) CQLQSNRAC(1) CTPESKAMC(1) CTSMIPYQC(1) CDHTSAAKC(1) CNSPATRFC(1) CSQVPARSC(1) CPPTPHNMC(1) CVSARHPHC(1) CTTQRPPNC(1) CSHQGYNTC(1) CNPAWNPRC(1) CQGTPLTQC(1) CNAVGIYSC(1) CENEKLAYC(1) CTTASPLLC(1) CLQSPHQHC(1) CPISTDQQC(1) CPHDRSLQC(1) CSHESHRQC(1) CSSTNQPSC(1) CMNSPRPMC(1) CYQSHKAEC(1) CNKTPYTLC(1) CSSAEAFSC(1) CWSVNQPFC(1) CHFHSLNQC(1) CKSPTTTYC(1) CQSGTMLMC(1) CRAELQSLC(1) CTLDRPSAC(1) CQASISPAC(1) CALPSSQLC(1) CSDPTHAKC(1) CTPTHDNAC(1) CTANDIAPC(1) CAFPSVNSC(1) CTPSPTYSC(1) CPNNIARLC(1) CTPHTALAC(1) CSPNRETFC(1) CMGPSKHFC(1) CFNKSTAKC(1) CPTPTSLVC(1) CNQSSHPQC(1) CGPTKHSEC(1) CLPSAGKAC(1) CLEATQKHC(1) CQDSQVTHC(1) CSPLSKLAC(1) CQAKSQGSC(1) CPSSKSLLC(1) CTEHFPLQC(1) CSNLRLPHC(1) CTPSHPSRC(1) CNHQSQKNC(1) CTPNRNLLC(1) CPYTLKSVC(1) CPDSSSKSC(1) CPMVSAHLC(1) CPTSNNGTC(1) CDGVAKPTC(1) CSPAKYNSC(1) CPHVPALAC(1) CKEPSQNMC(1) CQSNPSNMC(1) CPLSNPQHC(1) CMSLSIPPC(1) CTKTQADSC(1) CNSTASRQC(1) CSSHKSTYC(1) CTQHIHTQC(1) CNPPSKLLC(1) CPKSSLLQC(1) CQPMNSLTC(1) CRSTFPLEC(1) CDFSSNKTC(1) CTLPTSNFC(1) CSLTKLSFC(1) CELGPSRSC(1) CSPTRGDDC(1) CTTHITKTC(1) CNDWTQNSC(1) CDLLQPNMC(1) CPVTPHGSC(1) CTASGTPQC(1) CTLSFANTC(1) CTQKSGPVC(1) CPSGALTRC(1) CPPQASLHC(1) CTYPQAKMC(1) CQPTPQRWC(1) CPRALYEHC(1) CPASEFSRC(1) CVVPRGSSC(1) CFGLTPSTC(1) CAPSRTSLC(1) CTTSKSLMC(1) CTHKSHTHC(1) CPNKDVAHC(1) CPSDHRSDC(1) CPNLPRANC(1) CVLHPSAKC(1) CKVDNPNHC(1) CPTHLKTHC(1) CTNPKGVLC(1) CWPTTQKTC(1) CRPYTPNTC(1) CQPAPGKQC(1) CTNTLTRLC(1) CAPKPALTC(1) CSTMRSPTC(1) CNTMDKRQC(1) CGQKAHPAC(1) CNHSTRLLC(1) CTPSWMRTC(1) CQVASPNRC(1) CPLTDPPMC(1) CTGMTPRQC(1) CHSEQSKSC(1) CPGTQHSLC(1) CQREPSNSC(1) CQATNFQAC(1) CPLDKSQQC(1) CQKNLLPQC(1) 180 Phage display (in vivo) PMID:23104667 Mouse spleen Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The library prepared in 500 μl PBS was orally administered into starved male SD rats using an oral zonde, before they were lethally anesthetised after 1 h of retention and again perfused via the left heart ventricle with DMEM supplemented with 1% heparin to remove circulating blood from the organs. 2104 CGLHPAFQC(26) CSMNQPTAC(15) CPHNTTASC(10) CSSHLSSSC(9) CGPIVHSNC(6) CNQLTPKMC(5) CQWSVYNNC(5) CNSLYPPQC(4) CHQVFGAYC(4) CHLPPTIHC(4) CKDPAQRHC(3) CPMMPSQAC(3) CEKTTPLRC(3) CLTNFHRTC(3) CKEASANYC(3) CTEMHPMRC(2) CLSHPPTMC(2) CKSETLLSC(2) CTGPHSFHC(2) CNQSHAQLC(2) CTPQSRLTC(2) CTHQYAKHC(2) CPKNPMSTC(2) CSHPNLAKC(2) CMNKSHSAC(2) CSLNDMRYC(2) CHLRAPLLC(2) CDQASRLVC(2) CLMLHAKSC(1) CPSRASMAC(1) CLDRPMSTC(1) CYSWHHSPC(1) CMQQTTASC(1) CDQLLHNMC(1) CDRQPPQPC(1) CLLSEPNSC(1) CKEPYTTEC(1) CTLHTIYQC(1) CTLNTIYQC(1) CSKYSERQC(1) CFSLPPRLC(1) CTQPKLSMC(1) CHTTPFGHC(1) CYATQPQRC(1) CATYSPDRC(1) CKLYTNPHC(1) CLDLAYANC(1) CHSTTSRSC(1) CHTTQPSVC(1) CARGAPPTC(1) CEESPKRYC(1) CSLMMPRLC(1) CNLSAHTTC(1) CGVFIDHSC(1) CDGKTLLHC(1) CGDNYTRQC(1) CSPRTPLSC(1) CQTHSYLGC(1) CNRTSPLMC(1) CADPLPLYC(1) CSTGPSPSC(1) CSNAMPDSC(1) CSAPRLPPC(1) CNMRSDNVC(1) CHSPFGNRC(1) CATPSQTDC(1) CGPSAPRVC(1) CTPMQWHLC(1) CSLPMTRLC(1) CNAQNHSQC(1) CQAGDTRQC(1) CYSYHEMSC(1) CTIANRAYC(1) CQEWSKKEC(1) CNSPRTAGC(1) CMKQLSSMC(1) CLPFVDNSC(1) CQGFDFPHC(1) CITKFTRSC(1) CPMNAPLLC(1) CTSLSPWSC(1) CPPSRHELC(1) CISAEPAAC(1) CSNPSLEGC(1) CNPAKPTYC(1) CHPKNNLQC(1) CKHNDSLAC(1) CPASHQPLC(1) CSPSALWPC(1) CTNPSHSQC(1) CIPSTNTRC(1) CTTGLRTHC(1) CSHTPQTWC(1) CTRPLHHSC(1) CSLPKVRAC(1) CQTWTLISC(1) CLPNNIMTC(1) CHPTSLQNC(1) CLHSGHGSC(1) CMETNLRNC(1) CRPHHSLQC(1) CPQNTAKAC(1) CTNSHTKPC(1) CLQLLSSIC(1) CHPRIHHSC(1) CLLNGTKLC(1) CTRTQGFSC(1) CTAFQSRAC(1) CTLADTLHC(1) CALASKTDC(1) CQLNYPHDC(1) CLGEVAPVC(1) CRPPLDRMC(1) CPDQSGHSC(1) CSGALTGRC(1) CIESMDSHC(1) CNALGHPTC(1) CRQLPPSMC(1) CKPTEGLTC(1) CASSHYNNC(1) CLKPGPASC(1) CPSTSLPRC(1) CILERSRDC(1) CARTMTQDC(1) CTVPKATTC(1) CPHLNPQMC(1) CSPLTPNIC(1) CNHKIHIRC(1) CGDGWTPSC(1) CTGMSPRVC(1) CSISYNSFC(1) CNWSQQSTC(1) CGSSHSFYC(1) CTPGRTAVC(1) CESPSHATC(1) CRGGASPTC(1) CVPGWNLEC(1) CLMSQWHLC(1) CMLPSRLTC(1) CTSTWSFRC(1) CWAFTGAAC(1) CTISVQPMC(1) CPLHPNNPC(1) CPIDKTRFC(1) CSGPLKKTC(1) CPTLAMITC(1) CEVALSKPC(1) CPTSHDMQC(1) CLHSFPNHC(1) CDQMPAWTC(1) CLGPIATGC(1) CLPSPWSQC(1) CTSSQLDSC(1) CVMQLRPTC(1) CVVSNSARC(1) CENKLSPYC(1) CQWSVYTNC(1) CYTWHTSTC(1) CNTSDSSVC(1) CSSMSMTSC(1) CSPLEQTHC(1) CQTVPTQTC(1) CTKMQPTTC(1) CMTQHTSPC(1) CASAGELAC(1) CPLNHALKC(1) CMLFNISEC(1) CALSLPLYC(1) 168 Phage display (in vivo) PMID:23104667 Mouse visceral adipose tissues Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) To screen peptides targeting visceral adipose tissue via transdermal route, male Wistar rats were anesthetized and the hair on the abdominal skin surface carefully removed with an electric clipper. Phage-peptide library prepared in PBS was spread onto the abdominal skin surface. 2105 AASGRLVDVYNSCVS CFCSCYGLIGQGDGV CGYRQLSLFLTCPSE CWCDGNLSNIDADLY DAFTPLLGVRQMRRL FLLFVPQSIDHLNGG FSCLTHAVSLPITGS FWCALRHVGSDHPLW FYCFCIIAYLCNMGL GTICVCTFFCMVRCA ICTFQDLLVVQTVSD IHVLSLQYNAAYVGT IYTCYIVVAWTFTVP LCYIRVRDSRCLSYM LSFWTFAFASVRMLN LVYLCSSAALHNSDS PFIVHGFDCERFGLG RSCVTLGCESWAVPR SCAVITYPSQDFNIS SNLLWCVGRFSMYCD VCCRLLWGYVLLFLL VCFVGVFPGMVNCDV VFSGLVALRLPGCTS VSLGDCVSYCGLSIG WLSGDFFGAVDLTVT YDLLGVYRLSSRFAW YGHFIFLIVLFLFVC 27 Phage display (common panning) 5 PMID:23026082 DOX-SAM Fanconi anemia group F protein, FANCF Not determined. Not determined. X15 T7 phage display library 2106 AHSSYGDRTKPT IENYGDRTKPTS NAPVDAFGLRTK DFGERTKHNLWV GYSDHFFGSRQK APNQEYFGPRHK DYGPREKLYQML YMESQYGPRSK DYGPREKLYQML FDNATWGPRMKA FGPRTK 10 Phage display (common panning) 3 PMID:23123213 Monoclonal antibody 6D10 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2107 AGTVSGPGPHHP HGQGPFWDPTLP 2 Phage display (common panning) 1 PMID:23270686 IgG from human Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2108 QTLYLGLGHMML(1) HSLRLGPGVLAS(1) KSISMGPGYTLY(1) HPAYLGPGDMRR(2) THLPVTLGPGTL(2) 5 Phage display (common panning) 2 PMID:23270686 IgG from human Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2109 CPRILGPGC(1) CPRILGPAC(2) CPRQLGPGC(1) CPRMLGPGC(2) CPRLLGPGC(1) CPRVLGPGC(1) CPRIMGPGC(2) CPTRMGPGC(1) CPTRLGPGC(1) CPLRMGPGC(1) CPKLLGPGC(4) G-P-G 11 Phage display (common panning) 3 PMID:23270686 IgG from human Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2110 HDSSKFPYVPSY(1) FNLHDSSKLPDT(1) HAATVDKIENMR(1) GLDSSKWPNWSL(2) DTADSSKYEPLM(1) SVSEGMKPSPRP(1) FSSDSSKVHYPN(2) 7 Phage display (common panning) 3 PMID:23178262 Anti-OmpU polyclonal antibody Outer membrane protein U, OmpU Not determined. Not determined. Ph.D.-12 phage display library (X12) 7 of 9 positive clones shared the core motif DSSK-P. 2111 CSYTSSTMC CPDVPHPAC CTISFDRYC CSNTSSTTC CTIENPRAC CLLSANTHC CPTDHRFPC CSTASTQAC CHTLHGSAC CKQAASQQC CNARTSLFC CEPEKPRTC CPSPSSTHC CSTSMSSTC CSNPTSSTC CKFHQSSTC CPTSMSSTC CMTQQSSTC CETSMSSTC CLHGPSSTC 20 Phage display (in vivo) 5 PMID:22955033 Male Sprague-Dawley (SD) mouse brain Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In the first-round male SD rats were anaesthetized and injected intravenously over 30s with 100L of saline containing plaque forming units of the library. The brain-associated phages from this fourth-round output were then injected into another animal for a final fifth round of in vivo selection. 2112 CIKSFNNAC(2) CPKSSGALC(1) CPYTRLNHC(1) CPGFAPFPC(4) CPFHTTLAC(1) CPAHPPKAC(1) CHLATPYTC(1) CLSAGHSAC(1) CTLSTHRSC(1) CPLSQNRSC(1) CEVPTHRQC(1) CTNHETPSC(1) CHNNHSRPC(1) CWGHPLGTC(1) CMDGPIQFC(1) 15 Phage display (competitive panning) 3 PMID:23254898 cGP antibodies α3(IV)NC1 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Bound phages were competitively eluted with α3(IV)NC1. 2113 TALATSSTYDPH(7) APWHLSSQYSRT(3) IEWSDNNNFDPR(1) DVFAKQFTLMLN(1) ALTLHPQPLDHP(1) KLWNLHPTQALW(1) GPKGIVQSLQVR(1) 7 Phage display (competitive panning) 3 PMID:23254898 cGP antibodies α3(IV)NC1 Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were competitively eluted with α3(IV)NC1. 2114 LAHKSRLYERHM(1) RLNNRARIILRA(3) NRLIMSQRGPFK(1) VDLNQPWYKTSY(1) IKLRAQSYSPSS(1) TQQRPNSPAYKI(1) DLMPSLPNYAPL(1) LKTWGTTKATAD(1) KLLFAIPLVVPF(1) AHPNTRLIQPSD(1) 10 Phage display (common panning) 3 PMID:23238955 Mink enteritis virus, MEV Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2115 KCCYSL(0.75) WYSWLL(0.10) WYAWML(0.08) 3 Phage display (common panning) 4 PMID:12168699 Extracellular domain of human ErbB-2, ErbB-2-ECD Not determined. Not determined. Not determined. f3-6mer phage display library (X6) Wells of the microtiter plates were first coated with streptavidin. Biotinylated ErbB-2-ECD was incubated with streptavidin-coated plates. Each selection was performed on the plates. 2116 SDLWEMMMVSLACQY GEAHIPTSEMREKGW 2 Phage display (common panning) 5 PMID:12413859 B-lymphocyte WI-L2 cells Not determined. Not determined. Not determined. X15 phage display library Panning against B-lymphocyte WI-L2 cells yielded one unique peptide-phage, denoted CHL8, that penetrated the cells. CHL8 contained two different peptides, SDLWEMMMVSLACQY and GEAHIPTSEMREKGW. 2117 TLTVLPW(5) LTVEPWL(1) LTVSPWY(6) LTVSPLW(1) LTVTPWL(1) LTVQPWP(3) LTVSPWT(1) VLTVQPW(3) LTVSLWT(2) PGVIPWN(2) LTYQTWP(1) ELYVSRL(2) NLYVASW(1) L-T-V-x-P-W 13 Phage display (subtractive panning) 5 PMID:12490548 The breast cancer cell line SKBR3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Prior biopanning on adherent SKBR3 cells, the libraries were pre-absorbed on HMEC and PBMC. These peptides share a major peptide core motif LTVXPW, where X is generally a polar uncharged amino acid. 2118 LTVSPWY(14) 1 Phage display (common panning) 5 PMID:12490548 The breast cancer cell line SKBR3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2119 WNLPWYYSVSPT(31) 1 Phage display (common panning) 5 PMID:12490548 The breast cancer cell line SKBR3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2120 CGYAMNWSDSC[1.920 ± 0.086][+] CSLNWSDRTIC[1.805 ± 0.003][+++] CLGPYSLNWC[1.702 ± 0.008][+] CSSLNFADVYC[1.918 ± 0.041][+++] CLYSLNWQDLC[1.643 ± 0.095][+++] CLFSLNAPGLC[1.850 ± 0.036][++] CGIADLNDASC[1.639 ± 0.016][+] CNGITSLNHTC[1.768 ± 0.051][+] CSLNREDGLLC[1.809 ± 0.004][++] L-x-S-L-N-W-E-D 9 Phage display (common panning) 2 PMID:12674507 Anti-MRP1 monoclonal antibody MRPr1 The multidrug resistance-associated protein 1, MRP1 Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA,Dot blot In phage ELISA, absorbance at 405 nm was determined. The pC88 control phage (expressing wild-type pVIII) gave a weak ELISA signal and its OD value was 0.002 ± 0.003. ELISA results were shown as average A405 of three independent experiments ± SD. It is considered that reactivities are as statistically significant when differing more than 3-fold from the observed standard deviation from the background signal observed with pC88 (wild-type pVIII). Besides, semi-quantitative evaluation of the intensity of immunostaining of phage spots on a nitrocellulose membrane was performed by Dot blot. Results were expressed as symbols like +, ++ and +++. Specific complexes originated by incubating mAb with library and secondary antibody at room temperature (RT) for 3 h were immobilized on streptavidin-coated dishes. 2121 LTLNWPGDM[1.772 ± 0.036][+] QSLNWPRDV[1.721 ± 0.019][++] LTLNYADNV[1.667 ± 0.039][+] LSLNWGSSA[1.752 ± 0.051][+++] HTSLGHEDT[1.921 ± 0.019][+] LSLNYAGAE[1.752 ± 0.041][++] ISLNFENYV[1.652 ± 0.023][+] PYSLNWPDE[1.695 ± 0.023][++] LTLNWSKTL[1.826 ± 0.013][+] AGLLELNGW[1.562 ± 0.068][+] ISLNIENEA[1.684 ± 0.027][+] WSLNWRSPQ[1.580 ± 0.042][++] YDLNWPQNA[1.643 ± 0.025][+] TRLNFEDAG[1.636 ± 0.021][+] LSLNHARSP[1.671 ± 0.027][++] LTLNMDSGQ[1.625 ± 0.039][+] LTLNHASSG[1.565 ± 0.054][+] ITLNREDHT[1.054 ± 0.004][+] LSLNHHNNW[1.676 ± 0.002][+] L-x-S-L-N-W-E-D 19 Phage display (common panning) 2 PMID:12674507 Anti-MRP1 monoclonal antibody MRPr1 The multidrug resistance-associated protein 1, MRP1 Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA,Dot blot In phage ELISA, absorbance at 405 nm was determined. The pC88 control phage (expressing wild-type pVIII) gave a weak ELISA signal and its OD value was 0.002 ± 0.003. ELISA results were shown as average A405 of three independent experiments ± SD. It is considered that reactivities are as statistically significant when differing more than 3-fold from the observed standard deviation from the background signal observed with pC88 (wild-type pVIII). Besides, semi-quantitative evaluation of the intensity of immunostaining of phage spots on a nitrocellulose membrane was performed by Dot blot. Results were expressed as symbols like +, ++ and +++. Specific complexes originated by incubating mAb with library and secondary antibody at room temperature (RT) for 3 h were immobilized on streptavidin-coated dishes. 2122 VSQTMRQTAVPLLWFWTGSL 1 Phage display (common panning) 4 PMID:14643452 H1299 cell line Not determined. Not determined. Not determined. IR20 phage display library (X20) 2123 MTVCNASQRQAHAQATAVSL(8/10) 1 Phage display (common panning) 6 PMID:14643452 Human adenocarcinoma lung cancer cell line A549 Not determined. Not determined. Not determined. IR20 phage display library (X20) 2124 CHDSLYRAC CLLPLAAPC CGTATLHWC CTTRGPSTC CIDPSLGLC CKHMHQHIC CTKSLLLAC CQTTPPFLC CLPHSQAHC CPLTPTTVC CQWAALRPC CPQELHPNC CAYPYWLYC CPFGMVHTC CTSPLPSQC CKTPIPKIC CDLREHTLC CPSYSTSYC CLSPISLQC CVTTLNLTC CKATMTATC CTSSTPKAC CHNPPRPQC CHQTNPNEC CKPTLPLSC CLSASTLMC 26 Phage display (subtractive panning) 1 PMID:14670181 Human colon cancer cell line HT29 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage pool isolated after one round of selection against HT29 cells was subtracted by five rounds of successive incubation with HCT116 cells. 2125 CKQHHVTEC CTHLMPLTC CPSNETTQC CNASLMSVC CVPEQRPMC CKDKDNLPC CELSRRPMC CKAESPMEC CPEKFRPMC CLTPEPQYC CPPHTLGLC CKLVPTHQC CSMSSHRWC CPSTAELAC CPMHQRPMC CAQPLKQNC CHNVRFPNC CPVSNLLQC CNGSYVWRC CVSNQIANC CHSSHTHQC CNWQPATHC CSRLDSPFC CSYDYAKHC CKNERAYLC RPM 25 Phage display (subtractive panning) 2 PMID:14670181 Human colon cancer cell line HT29 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage pool isolated after one round of selection against HT29 cells was subtracted by five rounds of successive incubation with HCT116 cells. 2126 CPIEDRPMC(8) CPIHDRPMC(3) CAIRDRPMC(1) CALRDRPMC(2) CAPQLRPMC(1) CPEWARPMC(1) CPLDKRPMC(1) CSLERRPMC(1) CELWQRPMC(1) CDLPMHPMC(1) CQPPMFYSC(2) CLPSKFSHC(1) CDHPVPWRC(1) CSLRTAAAC(1) RPM 14 Phage display (subtractive panning) 3 PMID:14670181 Human colon cancer cell line HT29 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage pool isolated after one round of selection against HT29 cells was subtracted by five rounds of successive incubation with HCT116 cells. 2127 CPIEDRPMC(6) CPIDERPMC(3) CPIHDRPMC(1) CALRDRPMC(6) CPMHQRPMC(1) CPLASRPMC(1) CPEKFRPMC(1) CVPEQRPMC(1) CDLPMHPMC(1) CQPQSQPMC(1) CQPPMFYSC(3) CFESQSRLC(1) CIHPVPWRC(1) RPM 13 Phage display (subtractive panning) 4 PMID:14670181 Human colon cancer cell line HT29 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage pool isolated after one round of selection against HT29 cells was subtracted by five rounds of successive incubation with HCT116 cells. 2128 PPWYSKTDL(2)[1.31 ± 0.02][+++] PPFYLKEVR(1)[0.70 ± 0.06][++] 2 Phage display (subtractive panning) 2 PMID:11169438 Anti-CD18 monoclonal antibody MHM23 Integrin beta-2 Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA,Dot blot In phage ELISA, average values (A405 nm) were obtained after two independent experiments of ELISA assays. The standard deviation of each pair of measures is reported. Data were expressed as means ± standard deviations. The absorbance of the wild type (wt) phage was 0.05 ± 0.00. Besides, positive clones were further tested in dot-blot. Results were expressed as symbols like ++ and +++. Biotinconjugated anti-mouse IgG Fc-specific secondary antibody was pre-adsorbed overnight with M13K07 UV-killed phage particles. Then specific complexes originated by incubating mAb MHM23 with library and secondary antibody at room temperature (RT) were immobilized on streptavidin-coated dishes. 2129 CPPPFYRRGEC(8)[1.04 ± 0.08][+++] CIPNAPIWIKC(5)[0.50 ± 0.01][++] CEPPFFRKLPC(3)[0.61 ± 0.07][++] CRLPPFSRKLC(1)[0.33 ± 0.04][+/-] CNHPIFYARWC(1)[0.24 ± 0.00][+] P-P-F-Y-R-K 5 Phage display (subtractive panning) 2 PMID:11169438 Anti-CD18 monoclonal antibody MHM23 Integrin beta-2 Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA,Dot blot In phage ELISA, average values (A405 nm) were obtained after two independent experiments of ELISA assays. The standard deviation of each pair of measures is reported. Data were expressed as means ± standard deviations. The absorbance of the wild type (wt) phage was 0.05 ± 0.00. Besides, positive clones were further tested in dot-blot. Results were expressed as symbols like +, +/-, ++ and +++. Biotinconjugated anti-mouse IgG Fc-specific secondary antibody was pre-adsorbed overnight with M13K07 UV-killed phage particles. Then specific complexes originated by incubating mAb MHM23 with library and secondary antibody at room temperature (RT) were immobilized on streptavidin-coated dishes. 2130 YCPRYVRRKLENELLVL(2)[0.089] PCLMYSKYNLLYPYPAY(1)[0.092] GCMMYRYPAPGNHHIQM(1)[0.034] RCCTRKRFLRWYGHSLL(1)[0.237] SCHFEALRRQTYMYFYV(1)[0.205] 5 Phage display (subtractive panning) 3 PMID:15182617 The ECs injured by ox-LDL Not determined. Not determined. Not determined. XCX15 phage display library ELISA Identification of positive phage clones was performed by ELISA. Absorbance at 450 nm was determined, reproduced from the graph and shown. If A values of phages are twice as likely to that of the negative control, these phages are positive ones. ELISA results demonstrated that all six phages were positive. Before incubation with the endothelial cells (ECs) injured by oxidized low density lipoprotein (ox-LDL), the library was pre-absorbed with normal Ecs. 2131 CVPELGHEC(73) CELGFELGC(1) CFFLRDWFC(2) 3 Phage display (subtractive panning) 3 PMID:15480432 Tumor-related antibodies Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The CX7C random peptide library were incubated with antibodies immobilized on protein G from serum. The pre-cleared phage library was then selected on affinity-purifed antibodies from ascites of the index patient. 2132 CHTFEPGVC(35) CAPSQTYHC(2) CPPEAMHLC(2) CKAMSWYAC(2) CTSPTNRSC(2) CHGKYFVSC(2) CNTHMTAFC(2) CPSTLTSSC(1) CPSTLTSSC(1) CKIPSAFAC(1) CSRESPHPC(1) CQSRLSLGC(1) CLDHFAPMC(1) CLDKKTTSC(1) CNMSPQLDC(1) CSQRQTLDC(1) CSTKLLHEC(1) CPHSPTSLC(1) CPQRHVNYC(1) CMMSQLAHC(1) CPMAHLEFC(1) CELIKESRC(1) CQPENLPTC(1) CPFKLSKHC(1) CASSLHTIC(1) CHPLRLPAC(1) CHQSVNKEC(1) CLQNPTPEC(1) CPTEAQLQC(1) CLFAQLGPC(1) CNQPTRALC(1) CTPRTQKAC(1) CIHFPSASC(1) CPLRIAQHC(1) PST, AM, SRS, LD, SPT, HY, SQ, MS, SS, AQL, CHTFEPGVC 34 Phage display (common panning) 3 PMID:15508130 Human MTC-derived TT cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2133 CHTFEPGVC(35) CHTFEPGVC 1 Phage display (in vivo) 3 PMID:15508130 Human MTC xenografts Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Female nude mice carrying pre-established human MTC xenografts following subcutaneous injection of TT cells were anaesthetized with avertin and intravenously injected with input phage library . 2134 FPSKTGAFVPFS NNSQKPAPVSPF QLSPVLARHNIS TKNMLSLPVGPG RHLPTLFAPTPT PRGVWTTMSLPH LPLTSLMPLGLH SVSLPYANLATH RLLDTNRPLLPY LPY 9 Phage display (subtractive panning) 5 PMID:15520208 Nasopharyngeal carcinoma (NPC) cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA,Competition experiment Of 44 phage clones reacted with NPC and NNM cells, 16 NPC-bound phage clones were selected by ELISA assay. When a peptide-competitive inhibition assay was performed to discover whether the synthetic peptide and the selected phage clone competed for the same binding site, the results showed that the binding activity of NPC cells with the L-phage (displaying RLLDTNRPLLPY) was inhibited by synthetic peptide (L-peptide, RLLDTNRPLLPY) in a dose-dependent manner. After being subtracted with NNM cells three times, the unbound phages were used to react with NPC-TW 04 cells. 2135 SVWSATFLSSSP 1 Phage display (common panning) 6 PMID:15670871 PED3 domain of PMCA1b Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2136 FRPNRAQDYNTN(24) 1 Phage display (subtractive panning) 6 PMID:15671538 Prostate Carcinoma Cell Line DU-145 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The library was added to 293 cells for a negative selection. The medium was collected and centrifuged, and the supernatant was transferred to DU-145 cells. 2137 CPWWSPHAC CPIWQPHYC 2 Phage display (common panning) 6 PMID:15680248 Extracellular domain 1 of plasma membrane Ca2+-pump isoform 4 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2138 YCPRYVRRKLENELLVL(2)[2.217/3.521] PCLMYSKYNLLYPYPAY(1)[2.029] GCMMYRYPAPGNHHIQM(1)[5.101] RCCTRKRFLRWYGHSLL(1)[2.107] SCHFEALRRQTYMYFYV(1)[2.117] 5 Phage display (common panning) 3 PMID:15806288 The ECV304 cells injured by ox-LDL Not determined. Not determined. Not determined. XCX15 phage display library ELISA Fourteen clones randomly selected from the third screening eluates were subjected to ELISA analysis. Absorbance at 450 nm was measured by using an automatic spectrometer, and the absorbance ratio of phage binding to injured ECV304 cells versus control ECV304 cells was calculated and reproduced from the graph. Positive phage clones were designated as those with an absorbance ratio of equal to or higher than 2. 2139 CPSLRLFNC(2) CTTLSTQIC(1) CEQTQTLQC(1) CTLTPKNIC(1) CTAPTVNLC(1) CSDLYLNFC(1) CMDTAYPHC(1) CQLVSYKEC(1) CFAVDEQEC(1) CFKQVRHAC(1) CLATRPPDC(1) CSDSLTPQC(1) 12 Phage display (in vivo) 1 PMID:16259560 Medullary thyroid carcinoma, MTC Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A Ph.D.-C7C phage display peptide library was injected via the tail vein into tumor-bearing RET-transgenic mice. 2140 CLGEPRAHC(1) CSRESPHPC(13) 2 Phage display (in vivo) 2 PMID:16259560 Medullary thyroid carcinoma, MTC Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A Ph.D.-C7C phage display peptide library was injected via the tail vein into tumor-bearing RET-transgenic mice. 2141 TAWSEVLHLLSR 1 Phage display (common panning) 7 PMID:16452157 PED1 of PMCA4 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2142 SAKTAVSQRVWLPSHRGGEP(22) KSREHVNNSACPSKRITAAL(1) WLSEAGPVVTVRALRGTGSW(2) 3 Phage display (common panning) 3 PMID:16569591 B lymphocyte A20 cell line Not determined. Not determined. Not determined. ON543 phage display library (X20) 2143 SAKTAVSQRVWLPSHRGGEP(8) KSREHVNNSACPSKRITAAL(1) WLSEAGPVVTVRALRGTGSW(4) 3 Phage display (common panning) 4 PMID:16569591 B lymphocyte A20 cell line Not determined. Not determined. Not determined. ON543 phage display library (X20) 2144 CLSDGKRKC CLDGGRPKC 2 Phage display (subtractive panning, in vivo) 2-3 PMID:16740707 C8161 xenograft tumors Not determined. Not determined. Not determined. CX7C T7 phage display library Normal prostate cells were incubated with plaque-forming units (pfu) of T7 phage displaying a CX7C peptide library.After being centrifuged, the supernatant (the normal prostate-subtracted phage library) was recovered and then incubated with cells derived from premalignant prostate tissue or prostate tumor. After subtractive panning, the library was subjected to two to three rounds of in vivo selections. 2145 GSPQCPGGFNCPRCDCGAGY(56) GTGSCGYGKLHTGYWCSYFP(4) VHMNCSWMRVSEGHPCESAD(3) NSSSCDTSVVRSTWACILQP(3) KYGLCRDETVFPSHSCTFTG(3) VYAQCGVNVRTGRGGCSRLM(2) NRLKCRAQATHSAAPCIRGY(2) RQNSCTYSDARRWALCWSGE(2) QLNSCIHSGDRAIRGCMDWV(1) VRAVCTTLKSRGHEECWSLQ(1) GRQGCYEHLWRLIAWCAIFL(1) LRMTCAFGVAQRSADCALSS(1) SIVNCSAALTDLPTRCGGNI(1) CGTRCVRCQNGPEASCEQPL(1) TPLFCGNHGRQPSPLCMKWD(1) FTTVCRQPRGHEAIVCGSGK(1) APSFCGTAMLGASRYCYSGP(1) GARECESGGPGMRKLCTQIN(1) NNRACFRTSKGNPAECPYLG(1) GSLACQNIVVCVKKQCNALC(1) KRASCQNPLFSNFFVCGLSE(1) LPNFCMDTSGRAGPLCMGSE(1) RHTVCRVSLSSVQGSCSHEY(1) H-S-[AG]-x(3)-I-R-G, SAA, S-C-T-[YF]-[ST], R-G-H-E-[EA], [TS]-G-R-[GA]-G-x(3)-[LM], R-V-S-x(5)-[LM], P-x-F-C-x(8)-P-L-C-M-x(2)-[DE], TVCR, DTS 23 Phage display (in vivo) 2 PMID:16885375 Human breast cancer tumor Not determined. Not determined. Not determined. X4CX10CX10 FUSE5 phage display The phage aliquot was added to physiologic saline, i.v. administered and then infused into the patient. 2146 RRKRMTILKSRM TRTKLPRLHLQS 2 Phage display (in vivo) 4 PMID:16967280 B16-F10 tumors Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) C57BL/6 mice bearing single, subcutaneous B16-F10 tumors were injected i.p. with the Ph.D.-12TM filamentous bacteriophage phage display peptide library. 2147 GTRQGHTMRLGVSDG[15.00, 0.89] GDVWLFKTSTSHFAR[12.50, 0.14] LWVFPAGGHLGRERS[6.92, NT] VMVPYSVRDSLFGSF[2.08, NT] SVGGWFRQHLVGTRM[5.00, 0.63] LASIVRWEQVPDALS[1.75, 0.06] SGVWAPTAGYDAGFH[2.00, 0.65] IAGLATPGWSHWLAL[1.67, 2.14] SHSDYRPAPWSGWML[4.00, 0.89] 9 Phage display (subtractive panning, in vivo) 4 PMID:16984734 PC-3 human prostate cancer cell xenografts Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Micropanning assay Phage collected from in vivo phage display selections were screened for specificity of binding to both PC-3 –derived tumor tissues and cultured PC-3 human carcinoma cells. In the first round of micropanning, phage were incubated with either excised normal muscle tissues or PC-3 tumor tissues. Bound phage were then eluted from tissues. The amount of eluted phage was then determined by phage titer, and the tumor-tissue-to-normal-tissue ratio was then calculated. All phage clones with a ratio of >1.5 were then used in a second round of micropanning with cultured cell lines. Phage were incubated with cultured HEK293 cells (nonrelevant control) or cultured PC-3 human carcinoma cells. Bound phage were again eluted and titered, and the cultured PC-3 cells/HEK293 cells ratio was calculated. NT denotes not tested. The 15-amino-acid fUSE5 library was injected into the tail vein of normal non-tumor-bearing CF-1 mice to preclear phage that bound to the normal vasculature and other nontumor antigens. The mice were then anesthetized, and blood was obtained and phages were isolated from the blood. Next, the precleared phage library was injected into SCID mice bearing PC-3 human prostate cancer cell xenografts. 2148 KTLLPTP SGVEFLH SKKDTHH TMAPSIK TQHQVTA VNDRNVK 6 Phage display (subtractive panning) 4 PMID:18416599 Mouse PDAC cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Thirty individual phage plaques were isolated and an ELISA was performed to identify the most selective phage for PDAC cells. Of the 30 phage clones analyzed, 16 phage clones (53%) had specificity for PDAC cells. We sequenced the top seven clones on the basis of ELISA and multidimensional analysis. Clones 27 and 5 share identical peptide sequences (KTLLPTP) and demonstrated ideal affinity and specificity for the target PDAC cells. Besides, the extent of phage clone binding and specificity for mouse PDAC and normal ductal cells was quantified via flow cytometry. Clone 27 sharing peptide sequences KTLLPTP was highly specific for mouse PDAC cells having a 112-fold specificity over normal ductal cells. Phage clone 15 sharing peptide sequences TMAPSIK was second in affinity with the rest having nearly identical specificity. The counterselection was done by incubating the internalized phage pool with normal pancreatic cells to subtract all clones that bind to both normal pancreatic ductal cells and PDAC. Then the internalized phage were ampli?ed in Escherichia coli, titered, and subjected to three additional rounds for a total of four rounds of positive selection on the PDAC cells. 2149 NSLTPCGRTRVTSC(2) NSLTPCRNPKKATC(2) NSLTPCGRTRDN(2) NSLTPCAREVTLLC(1) NSLTPCSREVTLLC(1) NSLTPCDTTIANCC(1) NSLTPCDRILSPSC(1) NSLTPCTPKKSGRC(1) NSLTPCTTSSLTDC(1) NSLTPCSTKRKPNC(1) NSLTPCGSNSLTPC(1) NSLTPCCKSLRPHC(1) NSLTPCTKPKRNNC(1) NSLTPCSTKRKPNC(1) 14 Phage display (common panning) 4 PMID:18564683 Anti-LPS monoclonal antibody 9D5 Lipopolysaccharide, LPS Not determined. Not determined. NSLTPCX7C T7 phage display library 2150 NSLTPCGRTRVTSC(4) NSLTPCAPRSSNRC(3) NSLTPCAREVTLLC(2) NSLTPCLLLAQTDC(1) NSLTPCNSKKIPTC(1) NSLTPCLGRISPPC(1) NSLTPCASNSLTPC(1) NSLTPCGRTRDN(1) NSLTPCAPQ(1) 9 Phage display (common panning) 4 PMID:18564683 Anti-EPS monoclonal antibody 4B11 Exopolysaccharide, EPS Not determined. Not determined. NSLTPCX7C T7 phage display library 2151 NSLTPCLGRVLANC(3) NSLTPCGRTRVTSC(2) NSLTPCPYPRKGSC(2) NSLTPCRFLRRTVC(1) NSLTPCVPKKNRTC(1) NSLTPCRGRTHPLC(1) NSLTPCVPKKNRTC(1) NSLTPCNICARQYC(1) NSLTPCVRNSLTPC(1) NSLTPCRGRTLHLC(1) NSLTPCRGRTHPRC(1) SLTPCSYVGKGSC(1) SLTPCDTKKNHCC(1) NSLTPCGRTRDN(1) 14 Phage display (common panning) 4 PMID:18564683 Anti-protein monoclonal antibody BPM Not determined. Not determined. Not determined. NSLTPCX7C T7 phage display library 2152 NSLTPCGRTRVTSC(2) NSLTPCAREVTLLC(2) NSLTPCGRTRDN(2) NSLTPCGPKRKATC(1) NSLTPCSGLLVANC(1) NSLTPCSKKNPGNC(1) NSLTPCGAESLTPC(1) NSLTPCGSESLTPC(1) NSLTPCCKSLRPHC(1) NSLTPCAKTRTAKC(1) NSLTPCRTKKSGTC(1) NSLTPCFTVARACC(1) NSLTPCKTRKSGSC(1) NSLTPCFTVARACC(1) NSLTPCGAESLTPC(1) 15 Phage display (common panning) 4 PMID:18564683 Anti-protein monoclonal antibody BPA Not determined. Not determined. Not determined. NSLTPCX7C T7 phage display library ELISA Peptide-displaying phage types were designated according to the selection procedure with each type of phage and antibody; T7/9D5, T7/4B11, T7/BPA, T7/BPM, and M13/4B11. Bound phage was considered specific if the absorbance at 492 nm was at least 0.05. The selected bound phages, T7/BPA were ELISA-positive for B. pseudomallei MAbs. 2153 SHSSSNSEQLNFVMKVVSRP(4) SHSSSGYVGPRLGSGIGSRP(1) SHSSSTVVMGRVWQYEQSRP(1) SHSSSGYVGPRLEVGDWV(1) 4 Phage display (common panning) 4 PMID:18564683 Anti-EPS monoclonal antibody 4B11 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Bound phage was considered specific if the absorbance at 492 nm was at least 0.05. Using a random 12 peptide M13 phage library, 7 from 10 selected bound phages (70%) gave positive ELISA results. 2154 PMSYRY TSSYRL VESYRV SYR 3 Phage display (common panning) 2 PMID:19377945 Anti-uPAR monoclonal antibody IIIF10 Human urokinase receptor, uPAR Not determined. Not determined. f3-6mer phage display library (X6) 2155 TARGLSARTAFAGLV PISGRHLSSSFRFGG GEPSVVISRSSRLSP 3 Phage display (common panning) 2 PMID:19377945 Anti-uPAR monoclonal antibody IIIF10 Human urokinase receptor, uPAR Not determined. Not determined. f3-15mer phage display library (X15) 2156 DLLWLL DLLWEL DLLWII DLLFEL DLLYLL DLLTIL DLLYDL DLLWSL DLVWLY WDFPAY WEFPAY PDWHEM PSWEDD EPPWVW GPWMLW D-L-L-[WY]-x-L, W-[DE]-F-P-A-Y, P-x(2)-W-x(3), x-P-x(3)-W 15 Phage display (common panning) 2 PMID:19377945 Anti-huPENK monoclonal antibody Not determined. Not determined. Not determined. f3-6mer phage display library (X6) The phage-antibody reaction suspension was transfer to the streptavidin plate. 2157 YPLQPILAFPLA TLQSILPPRLWS VMHPSHFPVDAL WAITKPAPAAHP TPNLLVHMGVKL FMGHHMVEPWDV GFSGHRLFDLSS NPPLYPAAQTYE 8 Phage display (common panning) 3 PMID:20423858 Aminopeptidase N Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA,Fluorescence Ten selected phages were verified by ELISA, and the absorbance at 450 nm was determined. Only one clone had an higheELISA OD value. Synthesized peptide P7 (GFSGHRLFDLSS) bound to the cell membrane of THP-1 cells as shown by immunofluorescence assay. The binding of the peptide P7 to THP-1 cells was blocked by CD13-specific monoclonal antibody WM15 at different levels. 2158 MWPTTTHSSPYH(9) FDWKYNLLAHPQ(1) 2 Phage display (common panning) 4 PMID:20423858 Aminopeptidase N Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA,Fluorescence Ten selelcted phages were verified by ELISA, and the absorbance at 450 nm was determined. Nine phage clones had higher OD values. Synthesized peptide P9 (MWPTTTHSSPYH) bound to the cell membrane of THP-1 cells as shown by immunofluorescence assay. The binding of the peptide P9 to THP-1 cells was blocked by CD13-specific monoclonal antibody WM15 at different levels. 2159 CRTIGPSVC CRDGISVIC CWKHSGLGC CYGGGGRSC CYLGRSRWC CWGRSDGAC CDRSAGDSC CGGARGFLC CKVPSRSMC CGSAWGFLC CRVGPRMTC CRPRGDALC 12 Phage display (in vivo) 3 PMID:21183793 CNS endothelial cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage library i.v. was administered into normal mice and recovered the enriched population of brain-homing phage after 3 rounds of selection 2160 QFPPKLTNNSML(18) SYDILKPNPQRL(5) SHGKPPSFSPYT(4) 3 Phage display (subtractive panning, in vivo) 6 PMID:21258297 Lewis lung carcinoma (LLC) tumor cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A random 12-mer peptide library was pre-selected in vitro on HUVEC (one round of negative selection) and on HBOEC (three rounds of positive selection) which resulted in an enriched phage pool, designated as mini-library. The mini-library was used for in vivo panning of tumor-associated phage (three rounds of in vivo panning). 2161 ASSGMCFTKKTVLC[1.334] 3 Phage display (subtractive panning) 3 PMID:21339361 Human factor H Not determined. Not determined. Not determined. ANL4 phage display library (X10C) ELISA Each clone was tested for binding to factor H by ELISA. The absorbance at 405 nm was determined, reproduced from the graph and shown. The absorbance of phage CK, unrelated control phage from library ANL4, was 0.094. The libraries were prescreened by incubating the library in BSA-coated wells for 1 h at room temperature; supernatants were then transferred to factor H-coated wells for binding. 2162 ASSYDVGYSHDCRF[1.235] ASSSRCTYDHWCSH[1.395] ASPSWCSYSHWCRH[1.332] ASSFKCDYSHWCLH[1.311] ASSNVCSYSYWCAH[1.102] ASSCMYSYWCTH[1.232] A-S-S-x(2)-C-X-Y-S-H-W-C-x-H 6 Phage display (common panning) 3 PMID:21339361 Human factor H Not determined. Not determined. Not determined. ANL5 phage display library (X8CX2) ELISA Each clone was tested for binding to factor H by ELISA. The absorbance at 405 nm was determined, reproduced from the graph and shown. The absorbance of phage CK, unrelated control phage from library ANL4, was 0.094. 2163 CKRGNSGSC CQRLVGFAC CLQASPNFC CNGSVRSFC CPGFGLAYC CFLFTFEAC CRGVLMRYC CRAFVVASC CQSHSAFVC 9 Phage display (subtractive panning, in vivo) 4 PMID:21829599 Human CD4+CD25+ thymic cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) CD4+CD252 cells were used for preclearing and CD4+CD25+ thymocytes were used for biopanning and selection of bound phage. 2164 YPHYSLPGSSTL 1 Phage display (subtractive panning) 3 http://www.zgylxtb.cn/oa/DArticle.aspx?type=view&id=200903007 NCI-H1299 non-small cell lung cancer cell line Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The lung cancer NCI—H1299 cell line was used as the antigen and MRC-5 was used as capture cell for subtraction biopanning from a phage display peptide library. 2165 CSPLSQSAC 1 Phage display (subtractive panning) 4 http://www.cqvip.com/QK/92593A/200712/26118140.html THP-1 cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) THP-1 cells stimulated by lipopolysaccharide were used as target cells and untreated THP-1 cells as absorbing cells. 2166 CRAKSKVAC(15) CSRPRRSEC(3) CNRRTKAGC(1) CSRPRRSVC(1) CSRPRRSWC(1) 5 Phage display (in vivo) 3 PMID:14667505 Dysplastic skin Not determined. Not determined. Not determined. CX7C T7 phage display library Two rounds of selection ex vivo and one round in vivo were performed. Forty-eight clones were isolated from the vivo round. 2167 RLDPTSYLRTFW HDSQLEALIKFM 2 Phage display (common panning) 3 PMID:23440926 Primary cultured chondrocytes Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 23 single phage plaques were randomly picked and sequenced. Alignment analysis revealed that the sequences of phage clones RLDPTSYLRTFW and HDSQLEALIKFM exhibited some amino acid sequence similarity, while the remaining sequences showed no clear similarity to each other. 2168 SWTYSYPNQNMD(3) DWTYSLPGLVEE(1) NWTWSMPTGNPA(2) GMTLRVLTNYTE(1) TLHVSENSWTYN(1) DWLWSFAPNVDT(2) TLSSQNPYMHKK(1) IDKQMMTSHKAI(3) QGMETQKLRMLK(1) GWYWETPLDMFN(1) GWVIDYDYYPMR(1) VTAENYQSFSVS(1) NNKMSSEMMSIV(1) STGTDLHSNARI(1) YEFDNLLNRTLW(2) EWTVNERTMWDL(1) 16 Phage display (subtractive panning) 2-3 PMID:23483995 The embryonic progenitor cell line W10 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Cell targeting peptide phages were selected from a 12-mer linear peptide display library by 3 rounds of selection against undifferentiated W10 progenitor cells which included a negative selection against adult human dermal fibroblast (HDF) cells at each round to remove peptides binding common cell surface markers. The specificity of peptide phage cell binding for the displayed peptide was determined by performing competition experiments with synthetic peptides to indirectly measure the ability of the free peptide to bind the surface of W10 cells. It is indicated that cell surface binding of the 4 selected peptides(YEFDNLLNRTLW, GWVIDYDYYPMR, GWYWETPLDMFN and DWLWSFAPNVDT) was sequence specific and independent of display on the phage particle. 2169 EMYRKPMHAQLR(1) SMKPAPPVQHQL(1) EMKPMAPITRYT(1) KPMPPMIRLVTS(1) GIAKKPSAPLQR(1) ASNMKPSAPMQR(1) K-P-x(2)-P-x(2)-R 6 Phage display (common panning) 4 PMID:23613856 Anti-topo-I antibodies, pt4 anti-Ap1-17 Abs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Panning of a phage display peptide library with anti-Ap1-17 antibodies from 2 patients identified novel, partially overlapping motifs KPxxPxR as the result of the alignment of specific phage clone insert sequences. 2170 IPIPRESIKPTW(18) DPFTRMSIKPTG(1) DLSPRLTIKPQR(4) DPMSRITMKPHI(1) LPWIRTTEKPQF(1) DHPQTRTAPKPV(1) DPHYRNSPKPDS(1) NYRETPKPTWPT(1) EFRSSVKPQHPL(2) AHDHRSSLKPTR(4) QGHMRQTAKPFV(1) R-x-[ST]-x-K-P 11 Phage display (common panning) 4 PMID:23613856 Anti-topo-I antibodies, pt4 anti-Ap1-17 Abs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Panning of a phage display peptide library with anti-Ap1-17 antibodies from 2 patients identified novel, partially overlapping motifs KPxxPxR as the result of the alignment of specific phage clone insert sequences. 2171 SGHQLLLNKMPN(1) RLLFRKIRRLKR(5) MDMRTTDIRDTS(1) RNHPATLTGTGG(1) GILSELGKALGG(1) GAPALSTPPLSR(1) 6 Phage display (subtractive panning) 6 PMID:23409125 The cell surface of E. coli Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In order to eliminate non-specific binding phages and to select peptides that bind specifically to E. coli, a subtractive phage-display approach where the library was first pre-adsorbed against S. aureus ATCC 25923 was used to eliminate phages binding to the Gram-positive cell surface. The remaining phage library was then used to affinity select for peptides binding to whole cell surface of E. coli ATCC 700928. 2172 CGSKTQAPC CGLPWLSTC CSTPHNLGC CLATGNQIC CPATLTSLC CHGAPNRLC CRPPIGAFC CTSQSQHMC CGPAVYMKC CQGDLPGYC 10 Phage display (subtractive panning) 3 PMID:23533448 Tumor necrosis factor alpha, TNF-α Anti-TNF-α polyclonal antibody Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) After negative selection on a BSA-coated well, the phage library was then incubated with TNF-α. 2173 CSPHTTIAC CEPFAGRSC CTSPLPGTC CSYEAHQTC CNNPLKSLC 5 Phage display (subtractive panning) 4 PMID:23533448 Tumor necrosis factor alpha, TNF-α Anti-TNF-α polyclonal antibody Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) After negative selection on a BSA-coated well, the phage library was then incubated with TNF-α. 2174 TVKYSTLVEWPY(8) GFRFGALHEYNS(16) NSIALINDTHKR(2) TWKFEPLGTFID(3) DTFHSPLVALVS(5) YTSHFPLETWPQ(5) ETVRQAEELFYV(1) SFRFFPLDMWPH(5) AYTTVPYMATLP(1) GGSIAASELEYY(2) HSTLKLGALTNY(1) GSFHSPLLAYVS(1) DTGHSPEPGKVP(1) GTFHSPLLDHKS(1) TVTFAPLRMWHP(1) GWLRSPSLLFSN(2) TYTFRPLYEPPL(1) GYTFQPLNEWAI(1) 18 Phage display (subtractive panning) 4 PMID:23365656 VPAC1 receptor VIP peptides Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage library was first added to Chinese hamster ovary cells (CHO-K1 cells). After the incubation,the supernatant was collected, and the CHO-K1 cells (and the phages bound to them) were removed by centrifugation. The supernatant containing phages was incubated with the blocked CHO-K1/VPAC1 cells. Among these 18 phage clones, GFRFGALHEYNS appeared to have the highest OD450nm and selectivity value, and it bound more effectively than any of the other clones. 2175 FTHEAFG(6) HLQHYQV(2) KWDATYT(2) VRWDATT(2) IGHGTWG(2) HASPGQG(1) NPHEQAG(1) KVWTLPR(1) RWDATRE(1) W-D-A-T 9 Phage display (competitive panning) 4 PMID:23308124 Anti-CFP10-ESAT-6 (CE) pAbs CFP10-ESAT-6, CE Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phage was eluted with 100ml solution of the free anti-CE antibodies (100 ????g/ml in TBS) to compete the bound phage away from the immobilized antibodies on the plate. 2176 NHQQQNPHQPPM 1 Phage display (common panning) 4 PMID:23350661 U251 glioma cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2177 YEVCGSDTVGC 1 Phage display (common panning) 3 PMID:23278311 Serine/threonine-protein kinase StkP Not determined. Not determined. Not determined. ANL4 phage display library (X10C) 2178 YTFGLKTSFNVQ 1 Phage display (subtractive panning) 3 PMID:23521800 Prostate adenocarcinoma cells, PC-3 cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection: A 10 ml solution amplified from the positive selection was added to a T25 culture flask and incubated with CO2 environment. Rounds of positive and negative selection were performed three times. 2179 PRSLHRP AKTPPSR GKPNQGR RRPADRP QSSNFDC KPAAHAV PGASVEW STAQSLK RISGLRV QLTLDAA NAPRKTP LRKESHA IRAGRGV EAKAILT RNQPRAL HQRSQTK 16 Phage display (common panning) 2 PMID:23503763 Recombinant serotype D NTNHA, rNTNHA-D Botulinum neurotoxin type A, BoNT/A Not determined. Not determined. X7 T7 phage display library The random heptapeptide-displaying phages that display the affinity to the NTNHA-D protein were screened by using the QCM-assisted method. 2180 KGHSLMP IPTLPSS 2 Phage display (common panning) 4 PMID:23313229 Anti-human KGF mAb Keratinocyte growth factor, KGF Not determined. Not determined. Ph.D.-7 phage display library (X7) 2181 LEPKGRYDDPWT ATRYDDIWASTA R-Y-D-D-W-T 2 Phage display (subtractive panning) 3 PMID:23499997 Anti-rabies virus IgG Glycoprotein G Not determined. Not determined. Ph.D.-12 phage display library (X12) Before screening, phages were incubated with non-immunized dog IgG to exclude non-specific adsorption of the anti-rabies virus IgG. Then the incubated phages were allowed to react with the positive IgG. 2182 DYPPYLDRYAHW TATTDIRIPYTE YFDNTAKNAPLL GSHSRIGSVSMY TPLHRYFHWDSP TSKALYTHYQFL HYQFSDVRHAWK LPKNNLGLDALL IDTFYMSTMSHS FMGPQESTLQRL MNQTTKFSIHGA DNDEPGSRYNWW SDKMWHPMYAAP SMTQASDIGRLW KQLHPIYVQNGT APWHLSSQYSRT TWKYDGHRFYQA AKTLYPLYSSPI ARYDAIWNSEAL AGTGDDIARMWR TDVFLPRYDRLW NVLFFRIMMRHW SKNLFTGYQSSS SKYASQNYFTPD SYIDRYHWWSTQ VIVPDRHSHWLT GNVDVRWIWGNF DNHEHRFWHWDS YFDNTAKLAPLL TEQSVSRYDHLW 30 Phage display (subtractive panning) 3 PMID:23499997 Anti-rabies virus IgG Glycoprotein G Not determined. Not determined. Ph.D.-12 phage display library (X12) Before screening, phages were incubated with non-immunized dog IgG to exclude non-specific adsorption of the anti-rabies virus IgG. Then the incubated phages were allowed to react with the positive IgG. 2183 YSDEDLARVWRR KMSWDIGRVWNI S-D-R-V-W 2 Phage display (subtractive panning) 3 PMID:23499997 Anti-rabies virus IgG Glycoprotein G Not determined. Not determined. Ph.D.-12 phage display library (X12) Before screening, phages were incubated with non-immunized dog IgG to exclude non-specific adsorption of the anti-rabies virus IgG. Then the incubated phages were allowed to react with the positive IgG. 2184 FETLPSR NPLLSIQ LQGAHLR IPPLYFS QWPLMTT WRPPMLV HSQAAVP 7 Phage display (common panning) 3 PMID:23466786 Fibroblast growth factor 8, FGF-8 Fibroblast growth factor receptor 3, FGFR-3 Not determined. Not determined. Ph.D.-7 phage display library (X7) 2185 CTSQLLTHC CQHSKLSTC CIDLLHHKC CTGSLAISC CLGPAPHSC CQPAQSWSC CTPHRLAAC CRLHHTLSC CDPLKPSAC CHNPRANLC CTPHRLAAC CTQNPSSSC 12 Phage display (common panning) 4 PMID:23352850 NEP1–35 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2186 RTGNIVTAWK*X RRQTLSHQMRRP RRQSSRRMMRRI TNRKKRMISRMS MNRIPLPSITRR QHRLKPIMTRRQ RMRLNRRPKRMT MXLXRTRXXPKM RMLRMSRSSHRK RPLPRSKRNISR RRRKLRTQRLII KSHRRKRIRLLS SKRRNLQSRRKN TQRSKHRRRKNR HHRTRLRSMSRQ NSIHLMRLMRSI KSMQRRIKIINR R-R-x(7)-R-R-x 17 Phage display (common panning) 4 PMID:23352850 NEP1–35 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The displayed peptide with strongest affinity was synthesized as PepIV (RRQTLSHQMRRP) and its effect on axonal regrowth was tested in neurite outgrowth assay. PepIV could neutralize myelin inhibition and promote neurite regrowth efficiently. It was indicated that application of small peptides to block targeted myelin inhibitors may improve CNS regeneration. 2187 SPSIDTRYSRLG(3) CVSVGMKPSPRP(3) SVSVGMKPSPRP(3) MVSMDSSPRDRL(1) x(2)-S-x(7)-R-x 4 Phage display (common panning) 4 PMID:23338703 The human colon carcinoma Caco-2 cell Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After four rounds of selection, 30 individual phage clones were randomly selected and individually amplified. Of the 30 phage clones, 10 exhibited relatively high binding capabilities to Caco-2 cells. The novel SP-2 (CVSVGMKPSPRP) peptide, which could specifically bind to CRC cells, was selected in this study. This peptide has much potential to be developed as a more effective probe or marker for the early diagnosis of CRC. 2188 HLNILSTLWKYR(6) KLLTLSNWPNIR(4) HLPTSSLFDTTH(2) HLRLQDALPTRP(2) HLSWRDLQVQSY(1) HLPQNFTKLSP(1) HLPQNFTKLS(1) HLPQNFTKL(1) GSSHRLSWSQ(1) GHSQFASPSTAL(1) WH*GKISPLQVF(1) 11 Phage display (common panning) 3 PMID:23296614 Protein S100-B Calcium Not determined. Not determined. Ph.D.-12 phage display library (X12) 2189 MTVATPV NDVFRSG SISLEPL FYNVTVP FYNVTVP TLQFHTQ MHIAHSK MHIAHSK THWWRLW 9 Phage display (common panning) 4 PMID:23054431 BetaC1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2190 VLLAVWSVFHERSWL CLIVCLFIVRCGGCF GCNVWMCPDFCLPAR LGVVGFFPFDPSVLL SLYLPCVLPDLRFRM CGFAFYYVWLMVVMW QFFFSDDGSPPFEVG CSQGLLHSAVFALCF MLVCIFFFGMLLIMF FCVCFCGCTIMVLLL SSCNLVLAAHQFCGD LSLGSGFVVSYGVSG ASPLIVAAACSSYFL AFVSGADVNVCLHCA LFISIFVLAFCFALV SCLAVVLNCSGGLWS FCVSNGTTCVCIGVV TDVDFLFPNIHVSFG SVFCVSGSSSHWLIE FCVNNGTTCSVWRCD RVFFVIFLSDLIWPF SNAPCIVDMSSPLVH YSCDAYVGVRGVLCI GCVFILWGISSSHDA IFDALSLVCLSVQPS SASCFTGGMVRAHVR LHLIGLAASSGLSDL CVPSFGFVLPNRLAA LCCSVNLELSPSMGG VRPHPGLARSLPVAR ICNSVSLFVFGPYGA CDFLCVRSAVGTMVA CVYPAFTPFGQYTLL FLPGIGYASVLCCSG 34 Phage display (common panning) 1 PMID:23514038 Brz2001 DWARF4 Not determined. Not determined. X15 T7 phage display library Panning was performed using the QCM biosensor-based T7 phage display. The QCM sensor was immersed into the cuvette containing buffer and then allowed to fully stabilize. A T7 phage library was injected into the cuvette. Frequency changes, caused by binding to the Brz2001 immobilized on the gold electrode surface, were then monitored. 2191 TLHPAAD(7) NERALTL(5) SHSGYFS(5) APTEHWA(2) HATRPPM(1) TFNFPPV(1) AETHINQ(1) TMHGYDT(1) 8 Phage display (common panning) 3 PMID:23623136 Epoxy Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The results show that phage displayed peptide sequences exhibit high hydrophobicity, with low ratios of hydrophilic to total number of residues. The fluorescent characterization for the binding of TLHPAAD to a flat epoxy surface was investigated. Significantly, the surface with peptide-displaying phages shows much higher fluorescent intensity relative to the one incubated with M13 phages (without phage displayed peptides on the coat). Further, we investigated the localized binding of phage displayed peptide (TLHPAAD) to epoxy microstructures. The epoxy microstructures on glass were generated by using stencil masks to cover epoxy, followed by plasma oxidation. The phage displayed peptides were then allowed to bind to these epoxy microstructures, and the bindings were characterized with fluorescent characterization. Significantly, stronger fluorescent signals were observed localized on the epoxy patterns, relative to the glass substrate. This result clearly indicates that the microstructured epoxy in parts enhanced binding affinity toward phage displayed peptides over the host glass substrate. 2192 RDLHSNDVV(2) DKSSNDSIA(2) SNDGVWAIP(1) LFDRESNDW(1) YTDSNERTT(1) YRDSNEHQP(5) SND, SNE 6 Phage display (subtractive panning) 3 PMID:1720463 Anti-IL1β(163-171) monoclonal antibody Interleukin 1β, IL1β Not determined. Not determined. X9 PC89-based phage display library The library was incubated with the mAb and the mix was transferred to a streptavidin-coated petri plate. The 2nd round of biopanning was preceded by a pre-absorption step of the panned library on anti-mouse IgC antibody. Then the library was biopanned against anti-IL1β(163-171) mAb. 2193 YPVDRSTFW LASAGGQDN YPVDRSTFW NPIDHSARS NPIDATARS GPPVDPTSG DQWTAVGSK 7 Phage display (subtractive panning) PMID:11179365 Anti-TcTS Abs Trans-sialidase, TcTS Not determined. Not determined. X9 PC89-based phage display library The library was diluted to 100μl in phosphate buffered saline (PBS) and preadsorbed for 2 h with protein A-Sepharose 4B. Supernatants from four washings with 100μl of PBS were pooled, and 5 μg of\r\npurified antibodies was added. 2194 CPRVRSKPVVC CRRRLRGTWVC CSTDPAATDC CGPPVAPTSAC CVPLPLLRSKC CPRVRSKPVVC CPLSLRSKC CPRVRSKPVVC 8 Phage display (subtractive panning) PMID:11179365 Anti-TcTS Abs Trans-sialidase, TcTS Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) The library was diluted to 100μl in phosphate buffered saline (PBS) and preadsorbed for 2 h with protein A-Sepharose 4B. Supernatants from four washings with 100μl of PBS were pooled, and 5 μg of purified antibodies was added. 2195 HPLKQYWWRPSI(22) PIWWKHSGGPIL(1) YWWRDAPVSQGR(1) SYPTDKWWIKPG(1) W-W 4 Phage display (common panning) 3 PMID:11849705 E. acerulina sporozoites Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) PW2 (HPLKQYWWRPSI) disrupts the sporozoite pellicle, resembling the effect caused by most natural antimicrobial peptides. PW2 peptide was also effective against fungi and showed low activity against Toxoplasma gondii tachyzoites, but no activity against Trypanosoma cruzi, Crithidia fasciculata epimastigotes, and bacteria. Additionally, the parasiticidal concentrations of PW2 produced a very low lytic effect on mammalian and avian cells. The effectiveness against Eimeria sporozoites and the absence of adverse effects to host cells indicates that PW2 may be used as a model to generate new drugs for the control of avian coccidiosis. 2196 VQWWRPT(7) NWWRPLP(1) GKWWVFD(1) VPTKPWW(1) W-W 4 Phage display (common panning) 3 PMID:11849705 E. acerulina sporozoites Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2197 CPWWKTSKC(6) CPWWKASSC(1) CTPTWWRTC(1) CAPTWWKSC(1) CWWTSASRC(1) CSARWWQPC(1) W-W 6 Phage display (common panning) 3 PMID:11849705 E. acerulina sporozoites Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2198 SAGPYIDMMSIW KLQGNPSGPGYW ITGDRLAYFAYR DYLSNPKGPSHR DRAHEQEWFYKT TYRIGPYQGYHH FLPGIGDHRHSD GMRAGPMQNWTA FGPGSADFRHMN FSPGSADQKPHH GLRIGTGQSYWL 11 Phage display (subtractive panning) 3 PMID:23800339 Polyclonal RRi-11 plasma Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Three rounds of alternating positive/negative selection were performed. Positive selection used week 7 plasma from a protected animal, RRi-11. Positively selected recombinant phages were counter-selected with plasma from the same vaccinee but collected at week 0 (containing vaccine-induced Abs only). 2199 ESRQLYPGFQAF VIIGPPYQHWSM TLLRIPPGGVFM 3 Phage display (subtractive panning) 3 PMID:23800339 Polyclonal RTr-11 plasma Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Three rounds of alternating positive/negative selection were performed. Positive selection used week 7 plasma from a protected animal, RTr-11. Positively selected recombinant phages were counter-selected with plasma from the same vaccinee but collected at week 0 (containing vaccine-induced Abs only). 2200 TPFDLRPSSDTR(26) TDIRLRPLDKRI(4) RLKPRSDIKPIR(1) SDTPLIKRLDFP(1) RRSDLKPIRPLK(1) RRKIKPRRRIRP(1) TRIRPLRRRKPI(1) RRIPKLIKRRLR(1) 8 Phage display (subtractive panning) 3 PMID:23675691 Glyphosate Glyphosate receptors Not determined. Not determined. Ph.D.-12 phage display library (X12) Glyphosate-decorated glass beads were employed as binding targets and APES-coated glass beads as the negative screens during the phage library screening process. Phage colony containing TPFDLRPSSDTR(P1) has the highest binding ratio (19.2%). The second highest is TDIRLRPLDKRI(P2), which has a binding ratio of 5.2%. Both P1 and P2 contain a homology tripeptide sequence DXR (X = L,T, I, or K). 2201 WHWTYYW(9) WHWRAWY(3) DAFAWVP(2) ITYGFNP(2) WHWRAFF(1) AYWPSPH(1) TPLAQYF(1) LSFPYPR(1) SSMHTWR(2) WKSHQPP(1) THSLRFP(1) MPPQQYP(1) YPQPKHL(1) AHHSNLY(1) GPSVYSI(1) VWLKSAP(2) KYPPVHL(1) SWAQLPI(1) NPHKGMA(1) MRPPSPL(1) TTPTSRV(1) TLTNQLS(1) EPPPAQR(1) HHRIQTL(1) QASPQLS(1) GGASPIH(1) AHHMSDR(1) LPPNPTK(1) LPALQTV(1) WHWTYYW 29 Phage display (common panning) 3 PMID:23577599 Crystalline cellulose nanowhiskers, CNWs Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) From the single phage biopanning and ELISA analyses described, the binding specificity of the phage-displayed peptide WHWTYYW suggests that this segment might be a major determinant for interactions with crystalline cellulose substrates. 2202 YPHFHKHTLRGH(9) YPHFHKHSLRGQ(1) DHKPFKPTHRTL(1) FHKPFKPTHRTL(1) QSSCHKHSVRGR(1) QSSFSNHSVRRR(1) DFDVSFLSARMR(6) 7 Phage display (common panning) 3 PMID:23861727 Metuximab CD147 Not determined. Not determined. Ph.D.-12 phage display library (X12) 2203 KHSPVHI(4)[3.013 ± 0.186] LPLTPLP(1)[0.648 ± 0.081] IHSPVHI(4)[NT] YHSVVHI(4)[NT] HHTGVHL(2)[NT] NPLRVSL(1)[NT] NQDVPLF(1)[NT] HDTPAPP(1)[NT] MPSNTVR(1)[NT] SYSSFHR(1)[NT] SPTHGHD(1)[NT] THSIVHP(1)[NT] LHSPVHL(1)[NT] KHSVVHV(1)[NT] KHSPVHI 14 Phage display (common panning) 3 PMID:12413654 Peptidyl-prolyl cis-trans isomerase NIMA-interacting 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The ELISA plate was read at 405 nm and mean values and standard deviations were taken for the ten coated wells. Data were reproduced from the bar graph. The OD for control peptide NRPDSAQFWLHH was 0.599 ± 0.057. ELISA results demonstrate the phage borne peptide KHSPVHI, representing the consensus sequence, binds Par17 with the highest affinity and specificity when compared with other phage display derived peptides. All other peptides are non-specific and weak binders as the measured OD is comparable with the non-specific control peptide NRPDSAQFWLHH. The peptide consensus sequence XHSXVHΦ was enriched , where X can be any amino acid, and Φ is any hydrophobic residue. 2204 THSPVHVLAEHI(5)[3.273 ± 0.225] YHSDVHCANTCN(5)[3.351 ± 0.120] SHSVVHVTKNQY(3)[2.410 ± 0.172] WHTGVHISAKPG(2)[3.783 ± 0.120] ELTNAQIKLLWE(1)[2.090 ± 0.172] 5 Phage display (common panning) 3 PMID:12413654 Peptidyl-prolyl cis-trans isomerase NIMA-interacting 4 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Phage ELISA was utilized to measure the ability of the phage-borne 12-mer peptides to bind Par17. ELISA 96-well plate was coated with Par17 (10–20 μg/ml) and ten wells were coated. Absorbance at 405 nm was measured. Mean values and standard deviations were taken for the ten coated wells and shown. Data were reproduced from the bar graph. The peptide consensus sequence XHSXVHΦ was enriched , where X can be any amino acid, and Φ is any hydrophobic residue. 2205 SSFPPLL(8)[0.502 ± 0.163] LPLTPLP(5)[0.648 ± 0.081] GKPMPPM(1)[NT] 3 Phage display (common panning) 3 PMID:12413654 Peptidyl-prolyl cis-trans isomerase NIMA-interacting 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Phage ELISA was utilized to measure the ability of the phage-borne 7-mer peptides to bind Par17. ELISA 96-well plate was coated with Par17 (10-20 μg/ml) and ten wells were coated. The ELISA plate was read at 405 nm, mean values and standard deviations were taken for the ten coated wells. Data were reproduced from the bar graph. The OD for control peptide NRPDSAQFWLHH was 0.599 ± 0.057. ELISA results demonstrate that these peptides are non-specific and weak binders as the measured OD is comparable with the non-specific control peptide NRPDSAQFWLHH. 2206 SSFPPLL(13)[0.502 ± 0.163] LPLTPLP(1)[0.648 ± 0.081] GKPMPPM(1)[NT] 3 Phage display (subtractive panning) 3 PMID:12413654 Par17-GST Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Phage ELISA was utilized to measure the ability of the phage-borne 7-mer peptides to bind Par17. ELISA 96-well plate was coated with Par17 (10–20 μg/ml) and ten wells were coated. The ELISA plate was read at 405 nm, mean values and standard deviations were taken for the ten coated wells. Data were reproduced from the bar graph. The OD for control peptide NRPDSAQFWLHH was 0.599 ± 0.057. ELISA results demonstrate that these peptides are non-specific and weak binders as the measured OD is comparable with the non-specific control peptide NRPDSAQFWLHH. The phage library was incubated with Par17-GST in a positive selection. Starting from the second round, a negative selection was performed where the phage was incubated with the resins (in the absence of target). 2207 IVLPTWL YSTRHTD ADRSPLW YSSVHID YTSQTPR QTSITMY HRVLWHQ SFTYPSW ELCFKCS SDRSALW VSPGPTG SEWLWDH QCTLCPR YATPWQM TLIDPSR VLSDPPP RIPMQDL YVFPTWS ERMNHNP FYLPPWS ASQDITP IHTHPWY SHGSVHA GLGSVKP YIWPTWA VGFRLGL STTIALF SSPPKRL YAPPYQR STSELYG YDMSLIG QYMNQHA YDRSVLW HFVWTRS IELRGIY SDTTGLP EFTYPSW GWMMRFS QDRSILW ERVMSMM LAAKVHR MLDPRWE NFMYPSW TPMKISH AQSNSMH ILPVHVS VPPGTII QGLGTRG EPKTKHI HLSYGLN YIFPTWA YVFPTWM QFMSQVY SSGQHAF FIWPTWS TGSPQAR YGVMAPH RNDLPYT GQWSRIN GNNLVRT SSAQSSQ FKFTFAL SRYVDGY VPSRESW QLAMTYV MMNPIRN GELPGTA WIGIDTV LSINVVF EEWNWLR LSWTGKS GLFREGW MAQSSAA QYMYQLP DFWHHLR ADRSALF VPSFVPL QCLLTCA DSPLHAT QTYRPSY ANFWGES NFYLPTW WPISFLI YVFPTWM YSTNHID WKSPVLM VFGVHAH FNQPTWL HHSPMIG SLGKTVA AWPRHAA GIFKYFS NFVPPSW MGDIKHV TGTDATF CYMCQSL QYMFQTS TQGYATS NSGLHMF YPHRQVP YSRLHLD LPVERLN LLAVDAL HSPQSIA SGMTVLI WDPHWPK VVTIPYP LATRSSS GPHFELR RVLPSDE ETSSSIA HLQSSYQ SIRLHLG NGYINAM FYLPTWY EFHLGES MSNRDSE FTLPSWL KVTTMKP APPLSNL GSAQESH HHGAKHF TEFDVDH ITSPWPG VAKTYTS WGQNQSS LLGNIPP TSTTSRG YPFGVWT NHHAYPT ELALWST SWLSIFG TDNSARP HTAHVVT TVCCQTA FWFPPWE KFIIKAQ SVELQFG FYWPTWV MTQDITI SPLPWWQ WGGTLAR FSTVSQR RLVAQWP WTPMHDP GVQKASN DRLHMEP QFFWPAG WSSGVIF SLALMMR ETHHAHR LSHFSIV FIPRLSY SVTQSVI LMTPIMK KFVVKFN DHDRSWF TQHSYFL GNTYGFH IVMPPGP VSQSHLQ FRIPMID IFARTTM QHRALPT AVNTFML KGAYYQS VGLSIHI DKLYVTL TVRTGST GQDITAQ ADRSIMW THHHPDE QQFKSMS LIPTNTV LGTILHQ VHTGGLA IKISYPS MPGRGGV ATFGVWT NHVQGLP QFMSQMA IKLQPSV ETTQRPM GPYHAQP FTMPPWE FPFKWLA GASRAPA ADHVSHG VTSNFSH NSLTSYT MLERFLS HYFRING TFTYPTW HTHWFHT DPKGAYT YSSMHID IGQLVKL DSEHMTS WTWPSWA YTSQTIR KFWQGVT SKQDIIA SGMDRHE VTANTRE FSFTLTV AKVAITI FVRPTWH VSGTHML SPFFWEL TARSVWI VSTRHLN WNAPVPN NLDANLV KPNFDLH GAAGDAY TMTRNFM LRLNITW RIPMNDT YTGLQTQ CIFTCHD MDRSALW TISKWEE DRYVSWH EMRTERV DWNKAGD CHLYCEA GEWEYDH WNGMIGM EQTQARV YSTTHVD MPAHLNE SPFKWLA HISWRMM QEYHLAS LLGGAVA RIPMTDE NICIRCP QNWVPFN CYMCYQT SVVLNWN QFMYQPV YTTLHRD LANMSLL FVEPRSH SSGELWN MTDSHVS SYQMDYA AAKLLPH GRIGLQV QYMN*NF MPALVKF NLTHRGP LSLNMHD VLDFTLA 254 Phage display (competitive panning) 2 PMID:23826227 Anti-PAP sera sample 1, PAP1 Prostatic acid phosphatase, PAP Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage library was mixed with mouse or human serum. After incubation and washing, protein G was added to the phage-antibody mixture. 2208 AWFYPPW YVHPPWW LGYPWMQ NFRLPPW NPQWQTS YELPPWM FVRPSWI AFFGPSW WTRPPWE TFLPSWA YVFPTWS APPLFTP WTLPSWE NFQMPSW QWWNHEI GCCMHKN YINPSWW SLVDIWV FSLPPWA YVWPSWM GSLHFLV FTSPSWQ TYWPFPR SPLNIAR SYPWLIN KDLRDWF YVNPWEP FWEPPWG GDMHRVM TQVRPPW SFLPPWM WNWPTWE DLYYGRV GLWADNE FNSPSWS GFMAPSW LGSAWDV IIMPSWE GLNAALF APQMHLP TDGKSHL YVQPSWL WEKINHT AP*MHLP YLLPPWE HYLPSWH FSFPPWS LVLPSWL EQFYPAM ILWLLLW NYWPFPW VVWDRNP GYPWISN ESLNSFA TMWPFPS FVRPTWH HPLQVFV YFLPPWI NVLPPWL FELPSWM FVWPYWQ IVLPSWW APPMHII EFFFPSF VYAHTWL FVSPSWK FSMPSWQ ITLPTWL GFKEPSW TFRAPSW KAPHLES HFAFPSW FHRPSWD FTHPSWS VFVEPSW GGKLPLN SQLMPSW IWLPPWF FLLPPWN GWFLPSW WFINPTW AWWAPSW SFVKPTW KPVDLTA FRQPSWY AQWYPPA WVLPSWM FFDPSWN GFTLPSW FDFPPWA FNEPPWH VWMPSWA HEPTVMS RMVLPSW LTDHQRY NFILPTW ANLSQVL GFFLPTW SDHRHWI RFMVPSW VQMPASK HSLERLA FALPSWH GDYIPVK EDTKTSH FVFPSFH WTQQWYY LVYGDER VPSGKLH VVWDRHL ILLPSWE TFSYPTW VHNTPYQ QWWNLEI WHDPSWV DFIIPSW YPWINQP RFMDPPW FNPPSWG FINPTWA TWFAPTW VILPSWY VPGHHLM SVPSSEV FVMPYWA FKWPPWN LIYPSWA KSADHYY FSLQSFA SFMFPPW SFYEPSW TFTLPTW HSLEMLW SFQSPSW SYLRDLW HTHASGG AFRMPSW LVWDPWP FTQPSWQ RWVSPPW VLPSWAQ LIPVALT YWLPPWS EQFYPNW HVKAVET YEMPIPL GASRAPA NFTRPSW VVWDRSP MVVSSAN FTQPPWR FLVPSWH WFRMPSW QMEALMP SPPLQMF EFRSPSW IFSFPSW EVFWARL RPVDTPQ HSLVLPN QWWAGES KFQLPSW TFNPPPW YPWMQFS FLWPSWT QFRLPPW FNPMWQW FHYPSWR FTEPTWA VPRWFPP YYFPSWM YIVLPPW FFSPSWL RADYHAY YLGNYMI HLSTKGV TWWMREL FYSPSWT WPIFWH* QLTLPSW YSGQFNW NFAFPSW HFNEPSW ISTHYYL IPLNYNV FISPSWT NNNPQWQ FVAPSWN LVWPTWA WSWPSWV SLIQPSW QVLPTWS TYVYPPW TNPMW*M HSLEDVY LVNPPWA HMMRTNH LDFPFTS HVLPPWS WSEFRTL HSLEAIH SHYQYPD HKGFPMS LYLPPWQ LNLPTWM FLNPSWQ WFMPPWD DYLQNNA HSKPYLT FIRPTWG YHWATWE HFTMPSW YPEWGHN DQPWHWI YDMSLIG EFISPPW QVLPSWA HALEMIA WSTYRTA VTNFKTN LILPPWL WSWPSWN TRLYITD HIGQPRS VEDYMMR WQRLPSW YVRPPWS MVNPTWM LLWDPFP NVSLQMF QWHEPSW LWWSPSW NDAFASM YIMPSWS FEWPSWG FMNPSWK FIKPSWV HHIDSLW WSLHRTG YNWVQQS FRMPSWT NTVVLHL YVLPSWM HSLTIRN TYPWVTQ NWLPTWE FLNPSWE NWMWDAN AFIGPPW EIAMTTL LHHGVPV INLPSWM WTYPPWG HFTHPSC 254 Phage display (competitive panning) 2 PMID:23826227 Anti-PAP sera sample 2, PAP2 Prostatic acid phosphatase, PAP Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage library was mixed with mouse or human serum. After incubation and washing, protein G was added to the phage-antibody mixture. 2209 RVIAPSD WPVPYPL QWDDHW* QSPYPLN WPAREYF TLSGSRF TPQPYPL YETWEHR THSIFVY DWRNHWL AVSIVKR QWDDHWY RHEWLLH WLTPYPL AIPRLEE YHDAYPM YEGSIPH SPSPYPF VPSAYPL QANNTHM FWNTEML YNPWTHW NFRILSI THYRVNS KQPLHWY DLTPYPL DQPMHWF FSWRMMT DHVSWHY QWEDHWQ QDPYPMN DHTSWHP SGRIIGS SHVSWHT MHRMELR GHEPYPL DVPNWRT MLTPYPL YPNTDMT AWPRHAA VTITVQR CLAGGRP NPSAYPM THVSWHT DMPSQWG LRQPATG DMPLWWT EHPSWYP GGLIIGM ASNANLR WMPMRDR YPPFTDP GKPCEHC EFRLPQL TPQPYPL FNVGVKP QCDDHW* NHTPYPL LSKFPTS TVAQATS YRPSMNS SWAIRIY NWLKVGT TAWSVMK TVEYITN TFVSMPP RLPMVEP NALTGSA HYKPVYP IPHWVQT RPEPYPL QWEDHC* GQRVVIS TPQAYPL THSPYPM AHLSIIF QSSKFLH MSMSQWR QAALNRM FRPPMMD IGVHHWN WGLHTFS LPGFSVS KCCATHL DQPMHWI MSHMDSP YILRAGD CMQAHAM DHLMHQR QWEDHWE MQTRLMA HTTKPSL TVTPYPM ACLMCLT HPFNQLQ WTIVGPI KMEPYPL MHSPYPL QLDPYPL HLTAVLT MDILAFH KPNIAIL QSYLYGW QLEPYPL AYPLYPT EQPVHWS GVQKASN MTFTIAA DSFRVWP HLLNGKP AHDPYPL DQPSHWM NAHTYGI HGLHQFN VMRPVTA YPEKSPP LTHATMR APHRLTQ NMSPYPM GYSLHHR ELKIIQP FRLPMTD MFTRGQE RPPMNDL *IDPYPL ITSSHVW LRLPMHD HFGPRQY GPLMPPY VGTIPPL THVSWHI TSSVSLM DRSAFSS GYCAEDT VLGPYPL FRLPMND TCSAKWC GPTRYPL HLGPEQP ERDMRGI KDWPGRL QVTLSGH THWTRAD NKVIYYP LKTMPMI HLQWST* YGGFPMT VRSPFPM RPLPYPL HSLPHER YLSPYPL KATYIVL DRNMHIP ITKPEPN GYGKDWK MSYQIKR YTLSLSL NLEPYPL GMQPKPV WPGPYPL EHPSWAL ALTPYPL AHVPFHH DQTGGTN YARMLLS KMTPYPL TDWLVWE EAVPLRS TGSPSMA KQMPLYH SCPMCYW QEPYPLN KTSADMM GWSEHWS TDSRHVL DSWQKMR RPPMFDS HYKASLM HMTPYPM RIPMFDN MLMPYPL NQAKWVH SIPQASF WGLHTFR SLLMPLQ LSPTYLE RVPMNDG MTDSRAL WGVHQWR KFDGPMQ HLNANSK VYRSVEH HPFNRTS SF*TDHL VGMHTWN GVAHMTT DQPMHWL QWADHW* YGSAFTM SPMPYPL EHFSLHP THSPYPM MRLPMHD NHGPYPL SLMPYPL GQPVHWF HPVPYPM FPP*KHV VLTPYPF VVSASWL SVSETGG QDPYPFS WRLPMSD ELGSALL TIGRYPV KLSPYPL GDQGPNP DQPRHWM MPDPYPL VLGWNMN LQPHHCY ASIFTTW SWSNVSR SSNLTDR QSKYPLD LPSLQVL LDRQALM NHMLSAK NQPYHWL YGLHTFH SADHRML WRTHTWA DQPWHWI HYSYRSI NTFAFNH QYPYPLD QWEYHW* MTPLDAR NYWTQHR TNATTEL TRAPMHD HPFNRSH QWEDHCY QLTPYPL LHPHHWY YTFPFAS QWRSHWN WPLPYPM YEGGSMG RVPMVDG DPTPYPL NAKTSHM TDGQLRR SLEPYPI 254 Phage display (competitive panning) 2 PMID:23826227 Anti-PAP sera sample 3, PAP3 Prostatic acid phosphatase, PAP Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage library was mixed with mouse or human serum. After incubation and washing, protein G was added to the phage-antibody mixture. 2210 VFIPYGH KVMQLHI QMQWAVT YRGISVN AYLFNPM HAPIGNN RDLATLQ TVNNRMY GVWMLPK LVSSYNF TSIMFRY SVSTNSM ETRT*TL QPGRIVQ GIGRVAL LVALRVG ASGSLCC VVPCAGL TYTNPGF ETGSNTI VMDLRIL ISLRIEM MPFWLAR MPYDRSR RIHVDLF TSIGVFS WNSVSRL FFWMTRI VSPVRQL MYQINVT SGEQYRI VPMQTRL SHTNPKS NILANDR ISTHYYL WTVVQTL H*EISVL YSYVHTV HYSATAL QPSYWSR YVQGSRL WPSFVPK TFAGYRV SISHRMY HVRANVP WNLGGGS ASKLSTG YQPVTYR QPDSNVA VIHGNPP SF*TDHL QQHHVHL VSPRAGM DRNPMSF DPAT*TV GSPQVKK RVQDLPH SFRLGSH TVHAAYM WGQLGIR HISKGMT AVQFL*T GVNIRGI GQRVVIS MVHDMLG GNGPLHW VGKMHAN LAAKSGT GTSPDVW HTLVTAR VSWSSSS ITWSEHP AYFNLLR ASHAVGV TEWLRFD SLKHPMY MSWKLLQ LNIKLTL QTGLSSS AFHVTRM AYPKADS VGTTVVK TGSLFLT IARFSAN HTPVLSN VSYGGRT SIYSWAV FRFSCRW NGLVYRE RYLDVWA EMYFTKQ ISLTEHP AQMPVNV YPHPRWT FSSLGSK WSTLTPP SAPDDSF TWIRSAL MLQTNYD DRHFNYL QLSNCCQ GSTSPIV LERMPNP MWRIDFR SQSTVLQ HNPGTHK SGLDLWT QTSGSKL QSQIPNS SLPLVYS HSWIMHT WCRGLVW TFTSYLT AIMHHMV ASLDRYN MIKYRVA SMLIPHP SLYP*IG VMQGVLF NHVS*VS TPHWYVK TEQYTLN QMRPQHT KLYQREP GSNSARS NATMSAS IYPKVGH STYKSML MSRGAFI IWFTFQD APGGVVR ERVLPRV VPLWYVK ATRPLNH EQQTIRL TWS*AQT QLWYKIN FPGVQTQ VLFPDIT TNKTFQL ALFSNGR EASTKLT AIRAPWS IGLSEHF SVPYKLL AAPHPAT YRITIPM YVELIFV TPHWYVR NTSTYLM TVLSAAL TWPEFTK YGFTSNR ELQIVPS LT*KAQY AKVDPED YPHPRWN NFDT*AL VSLTYHI GVAHMTT NAPWWGS SLTEKNW VANTHHG ECRPCAD NIVSNSS DPLLNGV YMTGSKP *TWRGAV VLYSEHP KIPLTSK SVFSFRL SSMRQMH ALVFESY WPTFIPR LDPHRWM TYTGFVS EFAISST WKSGLTY GLIVSPM NTTTLES FQRSTAH SHQDNLL HQRPFVP FMILPGN YEGSIPH WRHVP*T LKLLDSY QLAYSTR GVHYTSA IHMSPPL HVFPCCH TYPERSA IPRWYVK *TSHKYL LARITAE VFAKANP HVDGSLR YRFVNFQ QWRKTES MAFTNAM SFYLPIR LTCLHCN EHPTTQI MGAMARV AVMDKRN DLFARSH TLGTPWS SQVAMIR GSSNLNL EPLSIFS YYPPAHR NAFKMVR YQRPFNP VSQMRSP GQMPFRP SPPLGSS IVDTTWH TVVMYPG SPDPYLR QVYDRSK QGLSRPH SLFPMSL DTPLPPW WMTDRVV KPPMSKY SQGMNLR SHPWTSG WTTTWMY MFSTATA WMTPRWL LPTVRNS TPKWFVK QHYDRSR HWYDRDR HFFTLTI THLLIYM YPWSEQR ICWACSP TGSSWAP HLQTSYT GKLTNVR VGFYEHP RIPIH*N VLYTKNL DGGYWHV AISLTPR SLPT*GS RAPVNSV QEYNRER YPNTDMT LDNALDR DGKQVSI YVRLAVT NTQHVTS 254 Phage display (competitive panning) 2 PMID:23826227 Anti-PAP sera sample 4, PAP4 Prostatic acid phosphatase, PAP Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage library was mixed with mouse or human serum. After incubation and washing, protein G was added to the phage-antibody mixture. 2211 QNIYAGVPMISF EATNSHGSRTMG TVSWSTTGRIPL QLEFYTQLAHLI SMDPFLFQLLQL 5 Phage display (common panning) 4 PMID:23776619 BT483 breast cancer cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The surface binding activity of each individual phage clone was analyzed by flow cytometry. The five phage clones exhibited prominent binding to BT483 cells, with phage harboring SMDPFLFQLLQ (PC2) displaying the best reactivity. 2212 AAPSHEHRHSRQ(20) ALAHNPKTTHHR(15) ERPLHIHYHKGQ(8) TTHSKHHFPSSA(5) 4 Phage display (common panning) 3 PMID:23398942 Siliceous spicules Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding specificity/strength of the phages was assessed by the ELISA technique. From the ELISAs it is apparent that the clones HRH-1 (AAPSHEHRHSRQ) and HRH-2 (ALAHNPKTTHHR) bind better than HRH-3 (ERPLHIHYHKGQ) and HRH-4 (TTHSKHHFPSSA) (based on the optical density, data not shown) and those again better than the wild type M13 control phage. The binding specificity/strength of the phages was assessed by the ELISA technique. From the ELISAs, clones harboring AAPSHEHRHSRQ and ALAHNPKTTHHR bind better than those containing ERPLHIHYHKGQ and TTHSKHHFPSSA. 2213 MQGHELGFMKI(1) AETVESCLAKSH(6) MHQGPTSYNLQM(1) HPNGHILLELRQ(1) LSNTLQTTNKK(1) ELQMRALQRLQP(1) CLRNASPHLGCL(1) HTSDGLQLSNVQ(1) WPQGPDRAVLG(1) SVHLYSQMPTKK(1) SRMPTMIMEKGW(1) HPLLTYSASQKG(1) 12 Phage display (common panning) 3 PMID:23551658 Gamma-irradiated Salmonella spp. Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2214 SEAYKHRQMHMS(1) YYPHPPWSPQH(1) HIRWDVNHNSMS(3) VPWVTTYEPWGM(1) VSAARADFYAAM(1) SSYYPQLTAHRF(1) SAKTHPWSIWAY(1) WHNAWESWHYAN(1) KLPPSFDLTGAN(1) GPADNTSKHVIR(1) NRPDSAQFWLHH(1) SWMPHPRWSPQH(1) 12 Phage display (common panning) 4 PMID:23551658 Gamma-irradiated Salmonella spp. Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2215 ALDDYKAGDRGI[3.276 ± 0.060] RQGSWDYKGADR[4.005 ± 0.111] AMAGPDYKVADK[3.858 ± 0.111] QDYKHLDDLLYA[3.036 ± 0.703] YRTMSDYKLDDE[3.688 ± 0.454] D-Y-K-x(2)-D 5 Phage display (common panning) 3 PMID:23807489 Anti-FLAG M2 monoclonal antibody FLAG sequence Not determined. Not determined. SUT12 M13 phage display library ELISA Phage clones from the third round of biopanning were confirmed for their binding to anti-FLAG M2 monoclonal antibody by Phage ELISA. Average OD values at 405 nm were determined. Data were reproduced from the graph and shown as means ± SDs. 2216 NWYLPWLGTNDW(24)[++++] QWELPWLMQPPL(2)[++++] SPGLSLVSHMQT(2)[+++] QLPRTAL(1)[+++] GETRAPL(1)[+++] 5 Phage display (subtractive panning) 4 PMID:23671272 Monocyte-derived immature DCs (iDCs) Not determined. Not determined. Not determined. Ph.D.-12 and Ph.D.-7 phage library pool Flow Cytometry Binding of the phage clones to iDCs was analyzed by flow cytometry. +++, binding <80%; ++++, binding >80-100%. Results demonstrated that all phage clones, but not the control phage, showed strong binding. Prior to biopanning on iDC, the libraries were preabsorbed on human monocytes. A single peptide, NWYLPWLGTNDW (NW peptide), dominated the sequences. The NW peptide shares the motif NW-LPWL with peptide QWELPWLMQPPL. 2217 LDYRSWSPYATS(3) AGELSPNRSAFL(1) QIPPRPPLLTTL(1) HNVTWAALMANV(1) SYNTLTQIAKIR(1) NSTNPHESRPTS(1) Y-R-S-W 6 Phage display (common panning) 3 PMID:23554202 Norwalk virus virus-like particles, NV VLPs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2218 LDYRSWAPYATS(5) IQYRSWIPFSYP(2) FRSYESPNFRPP(2) YRSFDPWYPPVH(1) LTQQRSWSPYMP(1) THQNRQTADIPS(1) Y-R-S-W 6 Phage display (common panning) 4 PMID:23554202 Norwalk virus virus-like particles, NV VLPs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2219 LDYRSWAPYATS(4) IQYRSWIPFSYP(1) LSIRSYTSPQWQ(2) YRSFDPWYPPVH(2) Y-R-S-W 4 Phage display (common panning) 5 PMID:23554202 Norwalk virus virus-like particles, NV VLPs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2220 LPSWYLAYQKII(9) ISWADWTQRWRW(2) ALPTFGVISPFS(1) S-W 3 Phage display (common panning) 3 PMID:23554202 Norwalk virus virus-like particles, NV VLPs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2221 LPSWYLAYQKII(10) SHVSKLVYQSQS(1) S-W 2 Phage display (common panning) 4 PMID:23554202 Norwalk virus virus-like particles, NV VLPs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2222 LPSWYLAYQKII(11) LPSWYLAYQKII 1 Phage display (common panning) 5 PMID:23554202 Norwalk virus virus-like particles, NV VLPs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2223 WSSGMTPDTGAP APWHLSSQYSRT 2 Phage display (subtractive panning, competitive panning) 1/2/3 PMID:23315991 Appetite-regulating hormone Growth hormone secretagogue receptor type 1 Not determined. Not determined. Ph.D.-12 phage display library (X12) With the exception of the first selection round, a subtractive step was performed to remove any background-bound phages. For the subtractive selection step, amplified eluate from previous round of biopanning was suspended in PBST and added to beads that were not coupled with ghrelin. B-ghrelin (biotinylated human acyl ghrelin, Phoenix Pharmaceuticals, CA, USA) were coupled to streptavidin-coated magnetic beads. The bound phages were eluted with either 10 mM DTT (Sigma, St. Louis, MO, USA), acidic elution buffer, or with polyclonal antibody C-18 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) specific for ghrelin. A search in the mimotope database MimoDB 2.0 revealed that peptides APWHLSSQYSRT and WSSGMTPDTGAP have already been selected by other research groups using various targets. The same 2 peptides were also found in different articles and patents using Google search engine. They were considered false positive results. 2224 HHALRLE SHTPTKF ADTVPRH MEMKKTHPVLGA APWHLSSQYSRT 5 Phage display (subtractive panning) 1/2/3 PMID:23315991 Appetite-regulating hormone Growth hormone secretagogue receptor type 1 Not determined. Not determined. Ph.D.-12 and Ph.D.-7 phage library pool With the exception of the first selection round, a subtractive step was performed to remove any background-bound phages. For the subtractive selection step, amplified eluate from previous round of biopanning was suspended in PBST and added to beads that were not coupled with ghrelin. B-ghrelin (biotinylated human acyl ghrelin, Phoenix Pharmaceuticals, CA, USA) were coupled to streptavidin-coated magnetic beads. The bound phages were eluted with either 10 mM DTT (Sigma, St. Louis, MO, USA), acidic elution buffer, or with polyclonal antibody C-18 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) specific for ghrelin. A search in the mimotope database MimoDB 2.0 revealed that the peptide APWHLSSQYSRT has already been selected by other research groups using various targets. The same peptide was also found in different articles and patents using Google search engine. It was considered a false positive result. 2225 ANVQFML SINDLTF EQFLLIG QSYLWHM 4 Phage display (common panning) 1 PMID:23315991 Appetite-regulating hormone Growth hormone secretagogue receptor type 1 Not determined. Not determined. Ph.D.-7 phage display library (X7) 2226 SKQHPHH(1) SNSLISG(1) DQRLPFL(1) MHPHQTN(1) ALRPPPH(1) QSGSPTF(1) EPLQLKM(1) NLTLDPC(1) EPLQLKM(3) 9 Phage display (common panning) 1/2/3 PMID:23315991 Appetite-regulating hormone Growth hormone secretagogue receptor type 1 Not determined. Not determined. Ph.D.-7 phage display library (X7) A search in the mimotope database MimoDB 2.0 revealed that the peptide EPLQLKM has already been selected by other research groups using various targets. The same peptide was also found in different articles and patents using Google search engine. It was considered a false positive result. 2227 GPRPPSAPNMPL(5/10) 1 Phage display (common panning) 3 PMID:23643267 Serum from AS patients Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2228 CGQEISGLC[12.780 ± 0.633] CRIRMSAGC[6.543 ± 1.054] CWGGLSGLC[4.433 ± 0.727] CRVIVGPRC[NT] CRVFAGKRC[NT] CRVTRGHGC[NT] CIRVEAGSC[NT] CLRSGGLTC[NT] CEFGLSEVC[NT] CMIAGLC[NT] G(2)-L-S-G-L, ISGL, MSAG, RVTXG 10 Phage display (subtractive panning) 3 PMID:12629410 Human urothelial cells Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Binding assay Insertless phage served as negative control and defined αv integrin phage ligand served as positive control. Results are expressed as mean of triplicate wells relative to binding of insertless phage, which was set to 1. Phage input was e8 tu per well. Experiments were repeated 3 times with similar results. Data shown represented relative enrichment of phage ± SEM from triplicate platings and were reproduced from the graph. NT denotes not tested. Each library was incubated with MOLT-4 leukemia cells for 30 minutes on ice to deplete background and common cell surface binding phage (pre-clearing). 2229 SRXPXXX(0.13) 1 Phage display (subtractive panning) 3 PMID:23143954 NSCLC tumor cells Not determined. Not determined. Not determined. X7 and X11 FUSE5 phage display library pool Prior to NSCLC tumor panning, the negative selection was performed using the normal lung tissue of the patient. 2230 SRXPXXX(0.09) ARRPKLD(0.09) XRXPXXX(0.35) 3 Phage display (subtractive panning) 4 PMID:23143954 NSCLC tumor cells Not determined. Not determined. Not determined. X7 and X11 FUSE5 phage display library pool Prior to NSCLC tumor panning, the negative selection was performed using the normal lung tissue of the patient. 2231 ARRPKLD(0.33) ARXPXXX(0.42) SRXPXXX(0.18) XRXPXXX(0.60) S-R-x-P-x(3) 4 Phage display (subtractive panning) 5 PMID:23143954 NSCLC tumor cells Not determined. Not determined. Not determined. X7 and X11 FUSE5 phage display library pool Prior to NSCLC tumor panning, the negative selection was performed using the normal lung tissue of the patient. 2232 YHDYKLYDTSLK(1) DYKLSDKWAQHA(1) WDYKELDDWTMY(7) D-Y-K-x(2)-D-x-W 3 Phage display (competitive panning) 3/4 PMID:23583685 Anti-SFV FLAG mAb Genome polyprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) For round 1 of panning the phage library was mixed with the mAb. After incubation and washing, protein G was added to the phage-mAb mixture. 2233 EHSPAFIADPLY(1) IHTPAFNVNVRT(1) WNSPVMREFVSK(1) SHSPAFKSTSRL(4) ISPQHPLYSILS(1) AMTQWRATSNMG(1) YLSKGKTHSLHT(1) H-[ST]-P-A-F 7 Phage display (competitive panning) 3/4 PMID:23583685 Anti-SFV V2 mAb Genome polyprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) For round 1 of panning the phage library was mixed with the mAb. After incubation and washing, protein G was added to the phage-mAb mixture. 2234 MHTNYCRTCFLS(1) NGYLCRTCSLTT(1) TGPRFIGSMGPA(1) LQAQKIEVQNQI(1) KLHISKDHIYPT(1) VMRQETPLRVPD(1) HHNPQWIDSQAK(1) H-G-P-Q-C-[IR]-[DT]-[CS]-Q-L-P-T 7 Phage display (competitive panning) 3/4 PMID:23583685 Anti-SFV V3 mVAb Genome polyprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) For round 1 of panning the phage library was mixed with the mAb. After incubation and washing, protein G was added to the phage-mAb mixture. 2235 QQSFMEKWLEYE(7) GVPSFLDMWLLY(4) FNEHHRVHSVTR(1) YVDNSFKRVKAN(1) IVCDDDGRCPNV(1) S-F-[LM]-[DE]-x-W-L-x-Y 5 Phage display (competitive panning) 3/4 PMID:23583685 Anti-SFV V7 mAb Genome polyprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) For round 1 of panning the phage library was mixed with the mAb. After incubation and washing, protein G was added to the phage-mAb mixture. 2236 HPSDYLLTYPFK(1) TWGAWFDFQLHP(2) HPSDYTLPVHPR(1) HPTDYSLSYMYP(4) HPDDYTLRFNRM(1) YNLQSPKSKLFA(1) H-P-[ST]-D-Y-S-L 6 Phage display (competitive panning) 3/4 PMID:23583685 Anti-SFV V8 mAb Genome polyprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) For round 1 of panning the phage library was mixed with the mAb. After incubation and washing, protein G was added to the phage-mAb mixture. 2237 VHNMQKDMVLLQ(6) VHDFKLINMKNS(1) RDYHPMLDMTLP(1) KIALESYNLSSA(1) NGQHAPTVAYSK(1) HKANEWHMLPST(1) SVGLHPPLSRQQ(1) LDLSAFPHTGLP(1) LMKPSYDTAHRP(1) YHTTPQMNKMYS(1) V-H-L-H-P-x(2)-N-x(2)-L 10 Phage display (competitive panning) 3/4 PMID:23583685 Anti-SFV V10 mAb Genome polyprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) For round 1 of panning the phage library was mixed with the mAb. After incubation and washing, protein G was added to the phage-mAb mixture. 2238 FTNTDWLLLFGP(1) VAERSTEETFMG(1) NQTDHLFSTFIS(1) STYWMGLIPAPI(4) HVRAYTAGILFE(1) HYTWMWDELSWY(1) GVTSFDFRRHTE(1) V-T-D-W-[LM]-F-[DE]-[IT]-F 7 Phage display (competitive panning) 3/4 PMID:23583685 Anti-SFV V11 mAb Genome polyprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) For round 1 of panning the phage library was mixed with the mAb. After incubation and washing, protein G was added to the phage-mAb mixture. 2239 HAIYPRH(4) GETRAPL(2) SILPYPY(2) SMYGSYN(1) KLPISSK(1) KDPGWSG(1) STASYTR(1) IQSPHFF(1) YLTMTPT(1) GPIIPRN(1) HAIYPRH 10 Phage display (subtractive panning) 4 PMID:23541699 Neural stem cells, NSC Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) After a negative selection against the plastic cell culture dishes, four rounds of panning against NSC were performed. 2240 KLPGWSG(9) HAIYPRH(1) GETRAPL(2) ALTPWAF(2) GKPMPPM(2) EQRGAPR(1) YSIPKSS(1) TDSLRLL(1) KLPGWSG 8 Phage display (subtractive panning) 4 PMID:23541699 Neural stem cells, NSC Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) After a negative selection against the plastic cell culture dishes, four rounds of panning against NSC were performed. To understand if the KLPGWSG peptide effectively binds to NSC we performed FACS analysis using antibodies recognizing NSC and the peptide functionalized with the fluorochrome FITC. These results strongly indicate a binding of our peptide to NSC. 2241 YLTQKPSPPYQG KIHYWPSTPTLT WTCQKAPCVARV FKMPQTMVMRTK GFNSAYKPQMRD LPYPQHPGSLGR FPPSWLAASNRP LPPQHPWDNSKH HSPVLKTPSTHA YSWHTDPKTLKR 10 Phage display (common panning) 3 PMID:23711778 Diamond-like carbon, DLC Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After three rounds of biopanning against DLC, 28 phage clones were randomly picked and their peptide encoding DNAs were sequenced. 18 peptide sequences were novel. Eight of the sequenced peptides, were longer than the 12 amino acids that would nominally be expected to be found in the PhD-12 library. 2242 VASPERPSPAFP(7) EASPERPSPAFP(7) AAPLGTHPSMHP(7) LPPLGTHPSMHP(7) SSATPNRLTWLL(7) YSHVHALSTYAR(7) VASPERTSPAFP(6) 7 Phage display (subtractive panning) 4 PMID:20211209 Anti-DENV2-NS1 pAb Non-structural protein NS1 Not determined. Not determined. Ph.D.-12 phage display library (X12) A negative selection procedure was performed in which the amplified phage was pre-incubated with the bead in the absence of the antibody. The supernatant was then reacted with the antibody in a positive selection. Twenty-five of 30 selected phage clones had significant enhancement of binding activity to DENV2-NS1-specific antibodies. 2243 RKKTPASRRPMR DLKPPIPSQSGP SPPYTYLPQKVQ HMGADPLQKRPS DLFARRPASSDW HFQMFHRPSGPQ SHQMAWPLEATS WHWTNWGKTSPA QHQHVNTLPERP HNWLYYLRTSTA HWRLPLLTSLMA NLTSLTQGSAML 12 Phage display (common panning) 2/3 PMID:19708689 Vitamin B12-binding protein Protein tonB Not determined. Not determined. Ph.D.-12 phage display library (X12) From BtuF-targeted assays, 120 unique peptides were affinity-selected from the Ph.D.-12 library. 2244 CHPTPTDIC CVHPYESHC CDSPLSNLC CHLTSMQMC CHLQTSSSC CWTPSPATC CYSTAVREC CTMRAVSAC CMASLSRSC CALKTNQPC CSHTQPYLC 11 Phage display (common panning) 2/3 PMID:19708689 Vitamin B12-binding protein Protein tonB Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) From BtuF-targeted assays, 81 unique peptides were affinity-selected from the Ph.D.-C7C library. 2245 TPLSLAPADQWL WVLAPADRARSL FDLAPADRNWGS QDLTLAPADYLA SMQAHLRLAPAD TQHLTMAPADSK MHQTLSPADLAT ALLCDWAPADCS 8 Phage display (common panning) 3 PMID:18653801 Anti-FhuA Fhu4.1 mAb Ferrichrome-iron receptor Not determined. Not determined. Ph.D.-12 phage display library (X12) 2246 SQLGVKYHMPGS QTGGLGVKYFRV QTQTLGVKYFRE YAQYGVKYKTET YFNDVQLGVRYH LPSTISVKYHST 6 Phage display (common panning) 3 PMID:18653801 Anti-FhuA Fhu6.4 mAb Ferrichrome-iron receptor Not determined. Not determined. Ph.D.-12 phage display library (X12) 2247 FPVHNLGVRYPG DPDQLGVRYHKA ELGVNYYSATSR SQTDLSVKYYNS 4 Phage display (common panning) 3 PMID:18653801 Anti-FhuA Fhu6.6 mAb Ferrichrome-iron receptor Not determined. Not determined. Ph.D.-12 phage display library (X12) 2248 HVTTTFAPPPPR(4)[0.316 ± 0.004, 1.979 ± 0.007] SVVPSKATWGFA(3)[0.303 ± 0.002, 2.220 ± 0.014] FKPSSPPSITLW(3)[0.327 ± 0.002, 1.645 ± 0.025] T-x(2)-F-[AK]-P-x(2)-P 3 Phage display (common panning) 4 PMID:21176936 Aminopeptidase N Transmissible gastroenteritis virus, TGEV Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Binding of the selected phages to pAPN was measured by ELISA. Ten selected phages and an irrelevant phage were incubated with coated pAPN. OD490 value of individual phage from three parallel wells is reproduced from the graph and shown as the mean ± SD of triplicate determinations. The absorbance of the irrelevant phage was 0.078 ± 0.011. Besides, the binding ability of the identified peptides to pAPN was investigated in ELISA by coating the pAPN as target protein. The TGE virion and IBV S protein produced from the same expression system were used as positive and negative controls, respectively. The OD490 value was reproduced from the graph and shown. The absorbances of the TGE virion and IBV S protein were 2.676 ± 0.014 and 0.472 ± 0.052, respectively. The chemically synthesized peptides block cell infection by TGEV through competition binding the viral cellular receptor. 2249 LLADTTHHRPWT(12/30) EHMALTYPFRPP(5/30) 9 Phage display (subtractive panning) 6 PMID:17125317 Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage library was preincubated with a graphite suspension to minimize the isolation of graphite-selective phage. Panning experiments were performed by mixing the recovered phages with the SWNT suspension. Biopanning was repeated six times with increasing Tween-20 concentrations (i.e., 0.2, 0.3, 0.4, 0.5, 0.75, and 1.0%). A new motif, X1THX2X3PWTX4, where X1 is G or H, X2 is H or D or null, X3 is null or R, and X4 is null or K, was identified from two classes of phage-displayed peptide libraries. 2250 ETAPLSTMLSPY(17)[1.924 ± 0.074] SVSVGMKPSPRP(14)[0.772 ± 0.028] TLHPSVLSYVLK(13)[2.025 ± 0.194] EVFWPLNAPRLL(11)[2.293 ± 0.179] ASTNVFARPMYL(8)[0.571 ± 0.063] APKYSLSDLYLN(7)[0.884 ± 0.153] TPPPRDASLSRW(7)[1.001 ± 0.096] SWQITYPISPRS(5)[1.628 ± 0.084] TALPNHWSDASP(4)[1.874 ± 0.123] SSFPQVIPLDYL(4)[2.293 ± 0.196] TDSYHVASARQP(4)[1.633 ± 0.327] YTFMPELTTPRT(4)[1.554 ± 0.074] NSFNYAPLLMPR(3)[1.633 ± 0.090] SGFFEPQHSHPL(3)[1.298 ± 0.055] TTFSPPSPLSSI(3)[1.237 ± 0.265] KTAPTPYALVHD(2)[1.385 ± 0.127] FNNHYPAAATYP(1)[0.720 ± 0.164] SWQIPYPISPRS(1)[0.910 ± 0.122] YTTWPFTSLQLD(1)[2.204 ± 0.401] ETAPPTPYSVMF(1)[1.940 ± 0.095] TTFNPLYLRLDT(1)[2.473 ± 0.090] TSPYHLLHAHLQ(1)[1.792 ± 0.084] SSFVALSISPSM(1)[2.393 ± 0.429] SPQTDGLVSTPS(1)[1.132 ± 0.166] SNFMHNTRIWSH(1)[NT] STYSHPLSLRPD(1)[NT] AWTWVLPSSIRA(1)[NT] AFMETTSQNAWL(1)[NT] QIEKISQHLDMH(1)[NT] TWNQPYIPPLYP(1)[NT] YASPPNPSLRLT(1)[NT] YHGLTPVRYVSV(1)[NT] ANLSSHSSPGDS(1)[NT] TTLQFTGQTNKT(1)[NT] HNIGTWGPKSHL(1)[NT] QIEESFVRGHTT(1)[NT] YTFDPQIRPAGL(1)[NT] YERSILPFSHVF(1)[NT] YPSVTFPTQTLL(1)[NT] SPWYMTPSPNTA(1)[NT] HISVINYTTKIS(1)[NT] MNVTVSGRLSGP(1)[NT] TIPYPFSLLNNP(1)[NT] PFLYSQVAWRS(1)[NT] YQEETPASSFSR(1)[NT] NSSQLAPYTTHR(1)[NT] HNGLPNFFQTRL(1)[NT] EAAPNFYPPLTF(1)[NT] DLFQFAFPLNTI(1)[NT] FTFSYAESVSYF(1)[NT] 50 Phage display (subtractive panning) 4 PMID:23792224 MDR GC cells SGC7901/ADR or SGC7901/VCR Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA In phage ELISA, unrelated phages (URPs, the amplified phages from original phage peptide library) and PBS were used as negative controls. Absorbance value was determined. Data were reproduced from the graph and expressed as mean ± SEM (n=3). The absorbances of URPs and PBS were 0.239 ± 0.028 and 0.093 ± 0.043, respectively. NT denotes not tested. Whole-cell subtractive screening from a phage display 12 peptide library was performed on MDR GC cells with a drug sensitive GC cell line, SGC7901, and immortalized gastric mucosal cells (GES) used as control. After four rounds of panning, 200 phage colonies were picked randomly and identified for MDR reversal capability using the MTT assay. GMBP1 (ETAPLSTMLSPY) could inhibit the tumorigenesis of gastric cancer MDR cells and might have the potential to re-sensitized gastric cancer to chemical drugsin vivo. 2251 KSFFLSH 1 Phage display (subtractive panning) 2 PMID:23714122 Pancreatic metastasis Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) GI-LI-N-derived primary tumor was used in the negative selelction. And corresponding pancreatic metastasis was used in the positive selection. Fifteen phage clones were selected for binding to the metastatic mass. 2252 YEGLISR 1 Phage display (subtractive panning) 3 PMID:23714122 HTLA-230-derived primary tumor Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Pre-selection on HTLA-230 cells was performed. Then selection on HTLA-230-derived primary tumor was carried out. 2253 HSYWLRS 1 Phage display (subtractive panning) 4 PMID:23714122 GI-LI-N-derived primary tumor Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Pre-selection on GI-LI-N cells was performed. Then selection on GI-LI-N-derived primary tumor from a different mouse was carried out. 2254 WSWPREL ALAAHKL 2 Phage display (subtractive panning) 4 PMID:23714122 GI-LI-N-derived primary tumor Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Pre-selection on GI-LI-N cells was performed. Then selection on GI-LI-N-derived primary tumor from a different mouse was carried out. 2255 SNYDTMLMHAKR[2.546 ± 0.154] TWNGTSNYDWPN[1.636 ± 0.118] SSNYHNKLHGEL[1.840 ± 0.131] AMLSPYDTNPWN[1.467 ± 0.067] ATDNYTTRAPRH[1.342 ± 0.101] ANPGQNPYEKRE[1.523 ± 0.120] QWFKSPLDVRHS[1.132 ± 0.096] GIGNFERYYGTA[2.123 ± 0.112] MPRRTRSIIISS[1.059 ± 0.086] SPRLTTYKASPK[1.800 ± 0.079] SNYD 10 Phage display (common panning) 3 PMID:23747319 Anti-NS2 4D4 mAb NS2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The affinities of selected phages to anti-NS2 4D4 mAb were measured by ELISA. Absorbance at 492 nm was determined, reproduced from the graph and expressed as mean ± SD. Phage ELISA results showed that the selected ten phage clones had high reactivity with mAb 4D4 (OD492 nm > 1.0) compared to a negative control of irrelevant mAb, the anti-porcine IFN-γ mAb (OD492 nm < 0.20). After three successive rounds of biopanning, phage ELISA results showed that the selected ten phage clones had high reactivity with mAb 4D4 (OD492 nm > 1.0) compared to a negative control of irrelevant mAb. 2256 CKSLEYSYC(18) CKSPENSYC(6) CTNTLSNNC(5) CTNTLNYNC(2) CKSTENSYC(1) CKSPENYYC(1) CKSPENYNC(1) CKSLENYYC(1) CKSMENSYC(1) CKYTESYNC(1) CKYLENSYC(1) CKSPENSSC(1) CNNTQSYNC(1) CTTPGNYNC(1) CKTRENSYC(1) CKSTQNSNC(1) CKNTENYNC(1) 17 Phage display (subtractive panning) 3 PMID:20709321 Pb2+ Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage library was first screened against uncharged monolithic column. Three rounds of main biopanning were conducted against Pb2+ as a target. The phage sample from the third round of positive selection was screened against uncharged monolithic column (post-negative screening I). After three rounds, the amplified phages were used in the next post-negative screening (post-negative screening II) against various other metal ions. Three rounds of negative selection against Cu2+, three rounds against Ni2+, two rounds against Co2+ and one round against Fe3+ were conducted. Sixty individual clones were sent for sequencing and 44 DNA sequences were obtained. 2257 TRWLVYFSRPYLVAT(9) 1 Phage display (common panning) 4 PMID:23806589 Anti-blood group A antigen monoclonal antibody Blood group A antigen Not determined. Not determined. f3-15mer phage display library (X15) Thirty-seven phage clones were chosen randomly and amplified. Eleven positive phage clones' exogenous DNAs were sequenced and the corresponding peptides were deduced. Among these eleven phage clones, nine clones displayed the same peptide - TRWLVYFSRPYLVAT, the other two phage clones displayed two different peptides respectively. 2258 NAHLQSDALPQT(7)[2.152 ± 0.045] WHWFRYPAPPPE(2)[2.152 ± 0.045] QVVSDNRWPTMK(1)[2.301 ± 0.033] WHYNWWLPAGPR(1)[2.626 ± 0.045] SISWWRWHWASV(1)[2.142 ± 0.087] HLYHYSYAAYWP(1)[2.274 ± 0.122] FHWSWYTPSRPS(1)[1.987 ± 0.100] 7 Phage display (common panning) 3 PMID:23707345 Anti-TTPI 4H11D10B11 mAb Triosephosphate isomerase, TPI Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Phage clones with mimotopes (1–7) were tested with ELISA to evaluate their reactivity to monoclonal antibody. TTPI was used as positive control and a M13 phage without insert (M13KE) and an unrelated mimotope (TsGST25) were used as negative controls of antigens. Additionally, anti-T. gondii mAb was tested in parallel as negative control (data not shown). Absorbance at 492 nm was determined, reproduced from the graph and expressed as mean ± SD. The absorbances of TTPI, M13KE and TsGST25 were 2.362 ± 0.120, 0.049 and 0.082, respectively. A total of 14 peptides were obtained after three rounds of panning of a random peptide phage display library (12 mer) using the 4H11D10B11 mAb. 2259 HHKTWHPPVMHL(8)[1.415 ± 0.061] SQWHPRSASYPM(2)[1.355 ± 0.035] WHP 2 Phage display (common panning) 4 PMID:23664850 Spike glycoprotein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Ten selected phages were incubated with the SARS-CoV S1 protein in ELISA plates to determine the specific binding activities for the protein. Three independent experiments were repeated for each individual. The OD492 value of tested individual phage was determined, reproduced from the graph and expressed as mean ± SD. The absorbances of phages with the same sequence were calculated as mean ± SD. The value of the negative control was 0.377 ± 0.071. The binding activities of 10 selected phages were assayed using ELISA. The results revealed that they had a specific binding activity to the SARS-CoV S1 protein. 2260 YGNAPDKPLSKR[1.327] TERAPDSPTSKS[1.443] ALLWEAPDQRLS[1.665] TERPPDSPASKS[1.572] STSPDFPLSSFY[1.603] HSHHTLTPDPPL[1.443] YAPTPPLSRIDP[1.567] ALHHHASRDAPE[1.340] APDPPLSR 8 Phage display (common panning) 3 PMID:23261158 Anti-CsdA A3G5 mAb Cold-shock DEAD-box protein A, CsdA Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The reactivities of selected phages with the mAb were assessed by indirect ELISA analysis. Absorbance at 450 nm was determined, reproduced from the graph and shown. The absorbances of the negative and positve phages were 0.225 and 1.799, respectively. The results showed that these 10 single phages expressed peptides that interacted with mAb A3G5 (OD450 nm > 0.3). These interactions were specific because no interaction was detected in the BSA control wells (OD450 nm < 0.08). 10 positive phage clones were randomly selected after three rounds of bio-panning and identified by ELISA. Eight of these clones were sequenced, and their amino acid sequences were deduced. 2261 DWLVWYG 1 Phage display (subtractive panning) 4 PMID:23888692 Toll-like receptor 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2262 CKTVPDNHC CDSPLTRNC CIGKFLYRC 3 Phage display (common panning) 3 PMID:23502767 MD-2 mimic peptide, NH2-FSKGKYKCV-COOH Lipopolysaccharide, LPS Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Western blot The Western blot (WB) analysis showed clearly that AP6 (CKTVPDNHC) and AP11 (CDSPLTRNC) indeed bound the MD-2 molecule. After 3 rounds of selection, 15 plaques were identified and their culture supernatants'ability to inhibit the production of cytokines was tested in PMA-induced U937 cells stimulated with LPS. Of the 15 clones, #5, #6, #8, #10, and #11 demonstrated good inhibition of TNFa and IL-6 production. We selected clones CKTVPDNHC, CDSPLTRNC, and CIGKFLYRC for further investigation as these 3 clones shows more inhibition effects than others. 2263 APTAVCNFFGQCPMEI APSKVCAHFDICYTLP APTTPCRKYFMCWEAG ALPKSCRILGTCQSIN 4 Phage display (common panning) 3 PMID:11179221 G protein β1γ1 subunits Not determined. Not determined. Not determined. f88-4 phage display library pool 2264 CGIHPWTKC(12/28) CGRWGHIPC(5/28) 2 Phage display (subtractive panning) 6 PMID:17125317 Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage library was preincubated with a graphite suspension to minimize the isolation of graphite-selective phage. Panning experiments were performed by mixing the recovered phages with the SWNT suspension. Biopanning was repeated six times with increasing Tween-20 concentrations (i.e., 0.2, 0.3, 0.4, 0.5, 0.75, and 1.0%). A new motif, X1THX2X3PWTX4, where X1 is G or H, X2 is H or D or null, X3 is null or R, and X4 is null or K, was identified from two classes of phage-displayed peptide libraries. 2265 CHTHNPWTC(3/15) 1 Phage display (subtractive panning) 5 PMID:17125317 Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage library was preincubated with a graphite suspension to minimize the isolation of graphite-selective phage. Panning experiments were performed by mixing the recovered phages with the SWNT suspension. Biopanning was repeated six times with increasing Tween-20 concentrations (i.e., 0.2, 0.3, 0.4, 0.5 and 0.75%). A new motif, X1THX2X3PWTX4, where X1 is G or H, X2 is H or D or null, X3 is null or R, and X4 is null or K, was identified from two classes of phage-displayed peptide libraries. 2266 CHTHPWTKC(3/11) 1 Phage display (subtractive panning) 4 PMID:17125317 Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage library was preincubated with a graphite suspension to minimize the isolation of graphite-selective phage. Panning experiments were performed by mixing the recovered phages with the SWNT suspension. Biopanning was repeated four times with increasing Tween-20 concentrations (i.e., 0.2, 0.3, 0.4 and 0.5%). A new motif, X1THX2X3PWTX4, where X1 is G or H, X2 is H or D or null, X3 is null or R, and X4 is null or K, was identified from two classes of phage-displayed peptide libraries. 2267 ATNAPRYTMQWS(5/15) EHMALTYPFRPP(3/15) 2 Phage display (subtractive panning) 5 PMID:17125317 Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage library was preincubated with a graphite suspension to minimize the isolation of graphite-selective phage. Panning experiments were performed by mixing the recovered phages with the SWNT suspension. Biopanning was repeated six times with increasing Tween-20 concentrations (i.e., 0.2, 0.3, 0.4, 0.5 and 0.75%). A new motif, X1THX2X3PWTX4, where X1 is G or H, X2 is H or D or null, X3 is null or R, and X4 is null or K, was identified from two classes of phage-displayed peptide libraries. 2268 LLADTTHHRPWT(3/10) 1 Phage display (subtractive panning) 4 PMID:17125317 Single-walled carbon nanotubes, SWNTs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage library was preincubated with a graphite suspension to minimize the isolation of graphite-selective phage. Panning experiments were performed by mixing the recovered phages with the SWNT suspension. Biopanning was repeated four times with increasing Tween-20 concentrations (i.e., 0.2, 0.3, 0.4 and 0.5%). A new motif, X1THX2X3PWTX4, where X1 is G or H, X2 is H or D or null, X3 is null or R, and X4 is null or K, was identified from two classes of phage-displayed peptide libraries. 2269 LPIFKVDFGDHSPFT(1) ARMFLLFLMRCIGCY(1) SHSFSVGSGDHSPFT(2) SHSFSVGSGDHSPFT(2) SHSFSVGSGSGDHSP(1) LSFFSCWLRRSFSLT(1) EVPRLSLLAVFLVVM(1) EVPRLSLLAVFLCNG(1) EVPRLSLLAVFLVAN(2) GRFLTGGTGRLLRIS(1) 10 Phage display (subtractive panning, in vivo) 3 PMID:23935860 Glutamate carboxypeptidase 2 EC=3.4.17.21 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) Poly-histidine tagged PSMA (PSMA-His6) is actually the target. A 15-mer library was screened against the soluble ectodomain of PSMA. In the initial step, the library was pre-cleared by intravenous administration in a mouse to remove phages that bound to vascular proteins thus clearing the phage pool of non-specific binders. After the third round of selection, three strong consensus sequences emerged: GDHSPFT, SHFSVGS and EVPRLSLLAVFL, where the former sequence appeared three times and the latter two sequences appeared four times. 2270 IDPVGWGNERTFQVP(8/8) 1 Phage display (common panning) 4 PMID:10975844 Monoclonal antibody TP25.99 β2m-free or β2m-associated HLA class I heavy chains Not determined. Not determined. f3-15mer phage display library (X15) Microwells were coated with avidin. The first round of panning was performed using 1.0e12 phage particles in TBS 50 and 1mg biotinylated mAb TP25.99 per well. The subsequent three rounds of selection were conducted using 1.0e10 phage particles and 0.1μg biotinylated mAb per well. We have mapped the conformational and the linear determinants recognized by mAb TP25.99 on HLA class I heavy chains. This information contributes to our understanding of the structural relatedness of distinct antigenic determinants defined by a mAb and of the conformational changes in HLA class I heavy chains induced by their association with β2m. 2271 QCTNFISDHECH(4/7) SCDGFYTGPACM(2/7) QCVETWNRIECK(1/7) 3 Phage display (common panning) 4 PMID:10975844 Monoclonal antibody TP25.99 β2m-free or β2m-associated HLA class I heavy chains Not determined. Not determined. LX-8 phage display library (XCX8CX) Microwells were coated with avidin. The first round of panning was performed using 1.0e12 phage particles in TBS 50 and 1mg biotinylated mAb TP25.99 per well. The subsequent three rounds of selection were conducted using 1.0e10 phage particles and 0.1μg biotinylated mAb per well. We have mapped the conformational and the linear determinants recognized by mAb TP25.99 on HLA class I heavy chains. This information contributes to our understanding of the structural relatedness of distinct antigenic determinants defined by a mAb and of the conformational changes in HLA class I heavy chains induced by their association with β2m. 2272 CWASNPSLC(1) CPYSNPSLC(1) CPFANPSTC(1) CNFSNPSLC(3) CEHSNPSLC(1) CWAANPSMC(1) CPFSNPSMC(1) CPYANPSLC(1) CSWANPSQC(1) CMFSNPSLC(1) CPFANPSMC(1) NPS 11 Phage display (subtractive panning) 3 PMID:16223774 Anti-CD20 monoclonal antibody rituximab B-lymphocyte antigen CD20 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) At each round, phage particles binding to isotypic and allotypic determinants of rituximab were removed by a preadsorption step on the rituximab isotype-matched infliximab. The epitope(s) recognized by Rp15-C (CPYANPSLC) is within the rituximab Ag-combining site. 2273 WPRWLEN(9) 1 Phage display (subtractive panning) 3 PMID:16223774 Anti-CD20 monoclonal antibody rituximab B-lymphocyte antigen CD20 Not determined. Not determined. Ph.D.-7 phage display library (X7) At each round, phage particles binding to isotypic and allotypic determinants of rituximab were removed by a preadsorption step on the rituximab isotype-matched infliximab. The epitope(s) recognized by Rp1-L (WPRWLEN) is within the rituximab Ag-combining site. 2274 EGHEHQTSWWEL(1) QDKLTQWPKWLE(5) HTSVERWPLWLE(1) ITPWPHWLERSS(1) TTTTSVWPAWLE(2) W-P-x-W-L-E 5 Phage display (subtractive panning) 3 PMID:16223774 Anti-CD20 monoclonal antibody rituximab B-lymphocyte antigen CD20 Not determined. Not determined. Ph.D.-12 phage display library (X12) At each round, phage particles binding to isotypic and allotypic determinants of rituximab were removed by a preadsorption step on the rituximab isotype-matched infliximab. The epitope(s) recognized by Rp5-L (QDKLTQWPKWLE) is within the rituximab Ag-combining site. 2275 TSPFTPVIGPLEHRS(1) STTNTPLVSHLEHRS(1) TGSVYSPTGLLEHRS(1) RETKLPFNVYTEHRS(1) XPPFTSAVGGVDHRS(1) APPFTSAVGGVDHRS(1) MDDDTERFPTHRSLP(1) RARDHSSTAQQEHAT(1) RAMAGRCRLLLSIGS(1) LEHRSPAE 9 Phage display (common panning) 3 PMID:10423148 Anti-gG2 monoclonal antibody H5 Envelope glycoprotein G, gG, gG-2 Not determined. Not determined. f88-15mer phage display library (X15) After three rounds of biopanning, individual phage clones were isolated and screened by ELISA to identify those which bound strongly to the antibody of interest, and those which gave a clear positive signal were sequenced. 2276 ATSLPPTEHPNMYQG(1) AGGYSPTEHAFHSPP(1) TSTPTEHTYPLEIIT(1) DPGTEHAGVPLRHSA(1) YGARPPEHLLYSRAS(1) SPLPEPPPEHRALVP(1) YMQPDPPPPLHAPDY(1) TRMPLPNHYEPPPRT(1) PPPEH 8 Phage display (common panning) 3 PMID:10423148 Anti-gG2 monoclonal antibody E5 Envelope glycoprotein G, gG, gG-2 Not determined. Not determined. f88-15mer phage display library (X15) After three rounds of biopanning, individual phage clones were isolated and screened by ELISA to identify those which bound strongly to the antibody of interest, and those which gave a clear positive signal were sequenced. 2277 ALSSQGGMSPEPTPL(2) VSSRPTTHYYLPEPL(1) TPESTPLLPPFPRSV(1) STNPEPLPFPAEELS(1) QKYAPETTPVSYLGA(1) HVLSSRPTTLALPLF(1) DYTPQTSLELPPESF(1) TPAQAYPALRSLIPW(1) TATTVTPRRTPYAPI(1) TPL 9 Phage display (common panning) 3 PMID:10423148 Anti-gG2 monoclonal antibody F11 Envelope glycoprotein G, gG, gG-2 Not determined. Not determined. f88-15mer phage display library (X15) After three rounds of biopanning, individual phage clones were isolated and screened by ELISA to identify those which bound strongly to the antibody of interest, and those which gave a clear positive signal were sequenced. 2278 QPTSKPTRPWLVSFL(1) NAETAFSLSSFPPSP(1) 2 Phage display (common panning) 3 PMID:10423148 Anti-gG2 monoclonal antibody H7 Envelope glycoprotein G, gG, gG-2 Not determined. Not determined. f88-15mer phage display library (X15) After three rounds of biopanning, individual phage clones were isolated and screened by ELISA to identify those which bound strongly to the antibody of interest, and those which gave a clear positive signal were sequenced. 2279 RGRGAG LGRRRS IRGRWA AGRSAF GRTDFW IKRNIT PRGRGG NKYARQ YHIKRS [GA]-R-x-[AG] 9 Phage display (subtractive panning) 1 PMID:7644467 Tissue-type plasminogen activator Plasminogen, EC=3.4.21.7 Not determined. Not determined. f3-6mer phage display library (X6) For positive selection, the phage library was digested overnight at 37°C by concentrations of t-PA varying from 10 to 100 μg/ml. The reaction mixture was then placed on ice, bovine serum albumin was added to 0.1%, and 100 μg of mAb 179 and 10 μg of mAb 3-E7 were added. After 30 min on ice, 100 μl of Pansorbin cells was added, and the reaction mixture was rotated at 4°C for 1 hr. The mixture was microcentrifuged for 2 min, and the supernatant was recovered to repeat the Pansorbin adsorption. 2280 VLRSAR QYLRYG KDARRA WLPRRA ILRAAY LRGRTA [GA]-R-x-[AG] 6 Phage display (subtractive panning) 2 PMID:7644467 Tissue-type plasminogen activator Plasminogen, EC=3.4.21.7 Not determined. Not determined. X6 fAFF1-tether C phage display library For positive selection, the phage library was digested overnight at 37°C by concentrations of t-PA varying from 10 to 100 μg/ml. The reaction mixture was then placed on ice, bovine serum albumin was added to 0.1%, and 100 μg of mAb 179 and 10 μg of mAb 3-E7 were added. After 30 min on ice, 100 μI of Pansorbin cells was added, and the reaction mixture was rotated at 4°C for 1 hr. The mixture was microcentrifuged for 2 min, and the supernatant was recovered to repeat the Pansorbin adsorption. 2281 GFARRA FGRRRT TRARRA [GA]-R-x-[AG] 3 Phage display (subtractive panning) 3 PMID:7644467 Tissue-type plasminogen activator Plasminogen, EC=3.4.21.7 Not determined. Not determined. X6 fAFF1-tether C phage display library For positive selection, the phage library was digested overnight at 37°C by concentrations of t-PA varying from 10 to 100 μg/ml. The reaction mixture was then placed on ice, bovine serum albumin was added to 0.1%, and 100 μg of mAb 179 and 10 μg of mAb 3-E7 were added. After 30 min on ice, 100 μI of Pansorbin cells was added, and the reaction mixture was rotated at 4°C for 1 hr. The mixture was microcentrifuged for 2 min, and the supernatant was recovered to repeat the Pansorbin adsorption. 2282 QRGRKA IVRRAE VARRAA [GA]-R-x-[AG] 3 Phage display (subtractive panning) 4 PMID:7644467 Tissue-type plasminogen activator Plasminogen, EC=3.4.21.7 Not determined. Not determined. X6 fAFF1-tether C phage display library For positive selection, the phage library was digested overnight at 37°C by concentrations of t-PA varying from 10 to 100 μg/ml. The reaction mixture was then placed on ice, bovine serum albumin was added to 0.1%, and 100 μg of mAb 179 and 10 μg of mAb 3-E7 were added. After 30 min on ice, 100 μI of Pansorbin cells was added, and the reaction mixture was rotated at 4°C for 1 hr. The mixture was microcentrifuged for 2 min, and the supernatant was recovered to repeat the Pansorbin adsorption. 2283 FTGRDI GFARAA IGRRAQ KAIGRM LGRKAT LKGRRA MRGRRG MRLRRA PFGRSA PYSRMA QLGRKA RGRRAR RGRSAG RRGRRA RYARRL RYARSA SRARKA SYLRRA TFARRA TVMRRA VARRAA WLGRRG YIGRRG [GA]-R-x-[AG] 23 Phage display (subtractive panning) 5 PMID:7644467 Tissue-type plasminogen activator Plasminogen, EC=3.4.21.7 Not determined. Not determined. X6 fAFF1-tether C phage display library For positive selection, the phage library was digested overnight at 37°C by concentrations of t-PA varying from 10 to 100 μg/ml. The reaction mixture was then placed on ice, bovine serum albumin was added to 0.1%, and 100 μg of mAb 179 and 10 μg of mAb 3-E7 were added. After 30 min on ice, 100 μI of Pansorbin cells was added, and the reaction mixture was rotated at 4°C for 1 hr. The mixture was microcentrifuged for 2 min, and the supernatant was recovered to repeat the Pansorbin adsorption. 2284 MGTTHLPYQFSL(3) 1 Phage display (common panning) 3 PMID:24040227 Listeria monocytogenes serotype 4b Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2285 KQATFDDYPVAH(7) MGTTHLPYQFSL(2) 2 Phage display (subtractive panning) 4 PMID:24040227 Listeria monocytogenes serotype 4b Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The first three surface biopanning rounds were performed against L. monocytogenes to select for phages binding to L. monocytogenes. The fourth included an initial subtraction biopanning step performed against L. innocua and using the supernatant derived from that for the subsequent biopanning against L. monocytogenes. 2286 KQATFDDYPVAH(3) KLHISKDHIYPT(2) 2 Phage display (subtractive panning) 4 PMID:24040227 Listeria monocytogenes serotype 4b Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) One solution biopanning was performed following the completion of the third surface biopanning round. The first three surface biopanning rounds were performed against L. monocytogenes to select for phages binding to L. monocytogenes. The solution biopanning included one solution biopanning against viable L. innocua and one against viable L. monocytogenes 4b. 2287 GPLATLHLPHKT(2) 1 Phage display (subtractive panning) 5 PMID:24040227 Listeria monocytogenes serotype 4b Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Two solution biopannings were performed following the completion of the third surface biopanning round. The first three surface biopanning rounds were performed against L. monocytogenes to select for phages binding to L. monocytogenes. Each solution biopanning round included one solution biopanning against viable L. innocua and one against viable L. monocytogenes 4b. 2288 ANAGPIMTTSLL(1) G*V*KPPQNVNR(1) GAKHPAQPHMMY(1) GAMHLPWHMGTL(3) GAMHLSVAYGYA(1) GEVDFNPR*DCA(1) GIIYDNPRKELN(1) GIIYSRGPEKLL(1) GIIYTLPAARYD(1) GIVFTLPALAHN(2) GKLFSSLDGL*F(1) GKLFSSPMDYDS(4) GKLYSHPLNNAK(5) GLLWTHPQTHGR(18) GMIWNEPKTWPG(1) GMIYVKPARPML(1) GNLFASPQKMHR(12) GPIFSAPT*TTI(1) GPIFSNSWGLIT(2) GPIFVNSDKGER(1) GPIHVAAFKNMT(1) GPILDMGFFNRE(1) GPIMSLPHRTVG(1) GPIMSLPTPTNL(1) GPINSKPSHMHI(1) GPIRDIGPVMDH(3) GPIVDSGGTHPR(1) GPIVSMPMPRLL(1) GPIWDNMPSRQV(1) GPIWSGRLIAQD(1) GPIYETIKTRTP(1) GPIYQQQNTILR(1) GPIYSTQHMKTS(3) GPLFDQGTQAYA(2) GPLHSSPLKISS(2) GPLISTPRHMNI(4) GPLVDLGPGDLR(6) GPLWTGQSQGSP(1) GPLYESRMPQNH(2) GPLYISSLTQLA(1) GPLYIVSHDTPR(1) GPVHSHPNDYSR(2) GQVYDVPYSRPK(2) GRIADLPPLKPN(3) GRIATLPDPTPR(2) GTDLD*AAAS*A(1) GTIFDYGPHGYA(1) GTIFDYGPPDMP(13) GTIWSQPGAISL(1) GVIWSDPKTASS(1) GVIYDKPA*KLH(11) GVIYDSHGPGRY(1) GVIYSKPNSVQL(7) GVIYSSDRDWRS(1) GVIYTDSLTRPH(1) GVMCKHPQTHGH(1) KLHISKDHIYPT(6) KQATFDDYPVAH(5) LYAKKPLLNPNR(2) NRPDSAQFWLHH(4) QSWPAAA*AFTS(1) TSMDSVSVIDLG(1) TSSQGDRLYVYK(2) TSWPSLSTSARS(1) VNLEHGYYHAPS(1) 65 Phage display (subtractive panning) 5 PMID:24040227 Listeria monocytogenes serotype 4b Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Two solution biopannings were performed following the completion of the third surface biopanning round. The first three surface biopanning rounds were performed against L. monocytogenes to select for phages binding to L. monocytogenes. Each solution biopanning round included one solution biopanning against viable L. innocua and one against viable L. monocytogenes 4b. 2289 ANATFHGYPTRS(1) FFPREYYSIEAP(1) GGNGPKGNNVVH(1) GKIWTEPPPPKP(1) GKIYNDPRMMSN(1) GKLFSSPMDYDS(1) GKLYSHPLNNAK(2) GKYHEKHNENMH(1) GLIWDLSWCSSK(1) GLLWTHPQTHGR(1) GMIWNEPKTWPG(1) GMIWSRPSAEKF(1) GNLFASPQKMYR(2) GPIATLPKGGGQ(1) GPIFSNSWGLIT(5) GPIHEAPSSPSG(2) GPIIA*TYPKRE(1) GPILDMGFFNRE(1) GPIMSLPHRTVG(1) GPIWSSIQTLPT(1) GPIYSSVKDGQR(1) GPIYSTQHMKTS(1) GPIYTDKSELGN(1) GPLATLHLPHKT(1) GPLFDQGTQAYA(1) GPLFITSAPPTK(1) GPLFSDPEPAKN(1) GPLISTPRHMNI(1) GPLVDLGPGDLR(1) GPLWTGQSQGSP(3) GPLYESRMPQNH(1) GPLYESSQVIRA(1) GPLYSSMASALA(1) GPLYSTNLPTRN(2) GPLYSYPFSMIE(1) GPSHNTLSPLLT(1) GPVHSHPNDYSR(1) GPVLDPLTPSTI(1) GQIYTTRDSLLG(1) GQKPT*NLDLKL(1) GQVYDVPYSRPK(2) GSIYAHPRWLKW(1) GTIMTLANTERP(1) GTIQVHPPAPAR(1) GVIWSDPKTASS(1) GVIYSKPNSVQL(1) GVIYSSDRDWRS(1) GVIYSTHDTRPY(1) GVLHSSPNHRWQ(1) GWHKHKSMSAPL(1) HLINTNAQIAQR(1) IQLEMGGTRFHR(2) KDDASPAWNSRH(1) KHMMINAYRMTE(1) KIHKTESTPAWF(1) KKGDV**L*LRR(1) KLHISKDHIYPT(4) KNLHVGSYPQPI(1) KPHAHKHNDYFL(1) KPHHPHKIPYTN(1) KPHNMTELHHKH(1) KPHYDHRLHQPI(1) KQATFDDYPVAH(24) KQSDVR*VSWWA(1) KRRAKQKTGEQR(1) LDLQTPGHKWSQ(1) LLPPT*ATVGAR(1) LSCTTSVACLQT(1) NVATKSSGHNMR(1) RQVRMHPLDSWS(1) RRKMKQTEKMKI(1) SKSRAKGKQAKN(1) SLNRKKRRTHAK(1) SLRQVNTHTWLT(1) TDAKMRAKFPGH(1) TQARCNEYPVGH(1) TSMDSVSVIDLG(1) VNLEHGYYHAPS(3) VNLQTGWYTMAS(7) VNSADWVVCDGV(1) VSLPMGFYSMNS(1) YLPISQTHNRNV(1) 82 Phage display (subtractive panning) 4 PMID:24040227 Listeria monocytogenes serotype 4b Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) One solution biopanning was performed following the completion of the third surface biopanning round. The first three surface biopanning rounds were performed against L. monocytogenes to select for phages binding to L. monocytogenes. The solution biopanning included one solution biopanning against viable L. innocua and one against viable L. monocytogenes 4b. 2290 CIQTTWSRC 1 Phage display (common panning) 5 PMID:15987688 Amyloid β peptide, Aβ40 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Thirty Aβ40 -binding clones were individually picked, amplified, and had their DNA sequenced. Most of the 30 clones thus obtained had different DNA sequences. The IQ peptide (CIQTTWSRC) may be a lead for the development of novel drugs to block the inhibition of nAChRs in AD. 2291 RRWVRYPVHLHSPIV(1) PPYHRFWRGHRHAVQ(12) HRISHFAHRYLARLH(2) WHWRHRIPLQLAAGR(2) GWHSLLHSRYHRIAA(1) FVWVRFHRLPRQIYT(1) WHKYFLRRPLSVRTR(4) VPMALNHGVYVMVSS(1) RKWFLQHRRMPVSVL(6) SGRRHLHRHHIFSLP(2) GWITFHRRHHDRVLS(1) RLHGHRSHRFTHVAQ(1) KRIWFIPRSSWYERA(3) MPLSRYWWLFSHRPR(1) RHLSHFKWLRSHGLD(1) RRFHFHSRMVAVDNS(1) 16 Phage display (common panning) 3 PMID:17804251 Transforming growth factor beta-1, TGF-beta-1 Transforming growth factor beta receptor type 3, TGF-beta receptor type 3, TGFR-3 Not determined. Not determined. f5-15mer phage display library (X15) Each selection was performed on the streptavidin-coated plates. In vitro and in vivo experiments were performde to valuate their capacity to neutralize TGF???21. A murine model of acute liver damage was used to test the in vivo activity of all 20 phage-derived peptides. As found in vitro, P17 (KRIWFIPRSSWYERA) was also the most active in vivo followed by P6 (PPYHRFWRGHRHAVQ), P4 (RLAHSHRHRSHVALT), P14 (SGRRHLHRHHIFSLP) and P11 (WHKYFLRRPLSVRTR). Other peptides such as P1 (DRRIFWWSLRSAPGA), P3 (RFFTRFPWHYHASRL), P7 (HRISHFAHRYLARLH), P9 (GWHSLLHSRYHRIAA), P15 (GWITFHRRHHDRVLS), P19 (RHLSHFKWLRSHGLD) and P13 (RKWFLQHRRMPVSVL) were without effect on the expression of collagen type I mRNA whereas peptides, P2 (HVRLHHYLRHRSLPN), P5 (RRWVRYPVHLHSPIV), P8 (WHWRHRIPLQLAAGR), P10 (FVWVRFHRLPRQIYT), P12 (VPMALNHGVYVMVSS), P16 (RLHGHRSHRFTHVAQ) and P18 (MPLSRYWWLFSHRPR) enhanced expression of collagen type I mRNA. 2292 CRMSRRSNC CMRNRPKRC CRRRLNRTC CRMRIRRNC CHPHPRPRC 5 Phage display (common panning) 3 PMID:20428769 Anti- FASN monoclonal antibody K1 Fatty acid synthase, EC=2.3.1.85 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A mimotope CMRNRPKRC against anti-FASN autoantibody can be used as an efficient diagnostic method of HCC. 2293 GNLFGRD(1) LSFPDWP(2) HSWLPYL(1) YSWTPH(1) QVLMMRP(2) YLIPPGL(1) FTPPVTL(1) TYLPGPL(1) TVSPGMK(1) DPPREIS(1) WHQTVPF(1) SISFWTL(1) IAKPGFW(1) WPKFHPS(1) GSSLDDS(1) SHLAPAL(1) TQSLSPP(1) IPLPQSP(1) 18 Phage display (common panning) 3 PMID:23845304 Anti-PCV2 Cap polyclonal antibody Capsid protein Not determined. Not determined. Ph.D.-7 phage display library (X7) 2294 VTAMEPGQ(28) AFNPEPGQ(11) VTALEPGQ(7) DTPPGWDQ(5) 4 Phage display (subtractive panning) 3 PMID:23885226 Rat mesenchymal stem cells, rMSCs Not determined. Not determined. Not determined. fd-tet X8 phage display library The landscape phage library was first depleted with culture flask to remove the phages that specifically bound to the culture flask. The depleted phage library was then incubated with target rMSCs. 2295 YEQDPWGVKWWY(8) HLSWLPDVVYAW(2) 2 Phage display (subtractive panning) 3 PMID:24046373 M2 macrophages Not determined. Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool For the subtractive pans, 2.0e11 pfu of the amplified eluate was first applied to receptor-cleared M1 cells. M1 cells were centrifuged, and the supernatant was applied to M2 cells. Phage was eluted from M2 cells and amplified. Subtractive panning was repeated three times, using the amplified eluate from the previous pan as the starting library for each round. M2pep (YEQDPWGVKWWY) shows selective binding and internalization in M2 macrophages but with minimal M1 macrophage interaction. We demonstrate this peptide's ability to identify murine M2-like TAMs within mixed cell populations isolated from mouse colon carcinoma tumors and to accumulate in TAMs after i.v. injection into tumor-bearing mice. We show that i.v. administration of M2pep fusion peptides with KLAKLAKKLAKLAK (KLA), a proapoptotic peptide, to tumor-bearing mice selectively reduces TAM populations and prolongs survival. 2296 NQYMSNGLVWAL(1) GDWPAGQMARSA(1) SPWSPPFWNDDM(1) QGPQTDSAPPRF(1) GFNTNPPSFPRP(1) GPLKAYILPPKA(7) AAHQPPAQSDFL(1) WSAPGLSSSSAP(1) LSSSAVTNNTSS(1) DSPTVAHNTSPT(1) HETVQHNKGWMI(1) IEHNGKAWRIPQ(1) EVAAFSMPGRFS(1) YVEPQEQSMPYL(1) NIQLELNPRHLI(1) SNSTREFNPNMF(1) HHQNTYANYPRH(2) WPHHHSRHNHNH(1) STHGWMNDRHHP(1) KSITSNDGFNTL(1) GSITTQTAIYFP(1) LTSAISPQHGEY(1) SHSFHTQERTTH(1) AVFSQLPRTPHL(1) SLPEAPIRQYQG(1) HVTMSWPQTAQN(1) SNAGGLMSRTWE(1) KIMPDSWAIKPW(1) NVMIDKHNVNGS(1) DSYMHAYSWRTK(1) SFQNSTLHGPVY(1) HIGHDNHLYPNR(1) NARVLHTGNESL(1) DRAMTPIYNPYI(1) TYTDNGYFKRST(1) TATIKSEAKSPS(1) YVEQVSTGKARS(1) RTRMRSINSPNL(1) SYYGKTDTADLT(1) VVDIRSQFANQQ(1) 40 Phage display (common panning) 4 PMID:24039747 Bacteriorhodopsin, BR Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2297 KVWSVPQLSHQL(1) KVWLLETSHISL(1) KLWYLNPPPGSF(2) K-[VL]-W 3 Phage display (common panning) 4 PMID:24039747 Nanodiscs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2298 MQLPKADARHPH YQITLPYRYEMP TFSWEFTGWWGQ LTFPVTTTPPAV MTHNMHGPNSEP HALTPIKYIPPG SSAYYYHYEYFH LSISPQHALVFA AMYHGHYTITRW 9 Phage display (common panning) 3-6 PMID:23830854 Membrane protein, M protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) To determine the impact of pepTGEV-M7 (HALTPIKYIPPG) on TGEV infection, three separate approaches were used: (1) TGEV was treated with peptide before inoculation of ST cells; (2) ST cells were treated with peptide prior to TGEV inoculation, and; (3) ST cells were infected with TGEV before being treated with peptide. These results indicate that pepTGEV-M7 is able to interfere with the ability of the virus to infect ST cells. The antiviral effects of pepTGEV-M7-treated virus were further investigated by IFA. Results demonstrated a dosedependent reduction in the infection of ST cells in the presence of pepTGEV-M7. 2299 NRPDSAQFWLHH YSAQFPLYTSWS WSEYDIPTPQIP GNLFASPQKMYR LNQSPQLQNKYR STFAGHVMTSVQ FPIKDPSNTFQY DFPISYRSWQIS RYWDTQLSPQML YQLQLPVWNDWA NMSHEMQLYALA WQLQLPVWSSSD ENMIVRLQSFQR EFFQYWEHIMWF NEPDFYAWQHFM WMWPSHMQEFSW VPFNWHWMQHYD YTVMDLQDYYSP GQLYMWMGLPIN QTTQAQTMLHHV SPPSTRVQNTAL QDWQQGSRVLWL TDYNLWTRAQFI WSSSAMQTIQTA MPAWEGPAQIQS MQRDNGPHKTLR FQCLRIPCVYAW ETWYSEQPWSML MLAHPSIYSSQP QFQWWYYSDQSE THWAKVSTDQHQ RIGTRRESKQNE QLPVRIQGNYLA LQQTYLKNPRSG MHDEVSVHQSHV NAHQKHAGAGAR QMAMRAHTEGKN YTSNTQSFTSSL WSFELPEQLFRG WDLTQRQRDLYF ANMEHTKHVTQA DKPGYMLLDSQP WRCRIELLMHQD STPSLLFYQVES QSTWWEIAGYTP MPNRPWIGILAG 46 Phage display (common panning) 5 PMID:23876241 Protein-glutamine gamma-glutamyltransferase Z Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2300 LGCDDTAVAHWPWVY SDAYDTAVAHWPGFI MPPNLIANYESALSR YESPGFLLSGYDDTG WAAHPSGPSIVPRSL 5 Phage display (common panning) 2 PMID:23939045 Anti-SMN monoclonal antibody MANSMA1-21 Survival motor neuron protein Not determined. Not determined. f88-15mer phage display library (X15) Three of these 22 mAbs were responsible for the selection process. These mAbs included two of the mostly widely-used for SMA studies, MANSMA1 and MANSMA12, and one mAb against the proline-rich region encoded by exon 5, MANSMA3. Biopanning with the remaining 19 anti-SMN mAbs alone failed to select any further phage clones. 2301 DHTLYTPYHTHP NHFGKFLDALAG 2 Phage display (common panning) 3 PMID:24073737 Bevacizumab (Avastin) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The most frequently selected mimotope, DHTLYTPYHTHP (12P), was conjugated to KLH and used to immunize BALB/c mice. Furthermore, anti-12PKLH antibodies induced the inhibition of tube formation, proliferation and migration of HUVECs. Our data indicate that the isolated mimotope 12P could potentially elicit specific antibodies against VEGF and result in the inhibition of human umbilical vein endothelial cells. 2302 NMSIATVP(4) ASCYIDDGIRYCTG(4) VSCNPFSWLPRCYA(4) DYCNLSVSWCYP(3) STCWLMFSMCRV(2) TPCEGSWFFCTL(2) LGCDVIYWSSGWGCKF(1) GSCSLSIVTIPCGV(1) VTCAAFSLFPFCYN(1) CHPGWPCFW(1) GGCNLSVLWCLD(1) NECILVNGDRWCIS(1) FPCWALPSQYLCRP(1) TACNLSVSWCEF(1) TNCGFLYLGCVI(1) FDCGDGSWRYYLDCVR(1) TSCRVLWIMTLCNA(1) EVCNNKLWPCFW(1) GGCGWLWVSCVI(1) CSPAWPCWW(1) LPCAGRSFPCYW(1) DRCLLWVSLCMA(1) CIPMWPCWW(1) 23 Phage display (common panning) 4 PMID:23922242 U-CLL mAb068 Not determined. Not determined. Not determined. M13 pVIII phage display library 2303 NSPDYASRLAAR(6) SNYAALLTGHHH(3) EPYHLKLAQRSQ(1) DNYAAALAQRAR(1) EGNWATALSSRA(1) APNYALLLNSRM(1) NSPTYASLLWGR(1) HFPSTTLRQVTT(1) NY, A-[ALST], L, R, P 8 Phage display (subtractive panning) 3 PMID:19690339 M-CLL mAb169 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection on BSA-blocked Protein A and Protein G beads was carried out in rounds 2 and 3 before incubation with the CLL mAb. 2304 WSYNKDLLDTLE(7) FSYSHESLRTLI(1) FSYTEFTLRQLL(1) FTYSHSQLHMLL(1) NFGYSQRWLSLH(1) APQYSLQLLQRL(1) FAFDEKLLRNLM(1) AWTYHPLRWNWL(1) F-[ST]-Y-[ST], L-[HR], L-[LI] 8 Phage display (subtractive panning) 3 PMID:19690339 M-CLL mAb183 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection on BSA-blocked Protein A and Protein G beads was carried out in rounds 2 and 3 before incubation with the CLL mAb. 2305 SPNFSWLPLGTT(9) HWPRPDDSFWRP(2) NHTFRLASPPVL(1) ASPANNPSPAWL(1) FSMPFRPITPVH(1) KDWRHPNWSGVL(1) NTKPSLPTISWP(1) HPPGIFPWTSKA(1) ATWSHHLSSAGL(1) LDSRPWPRLYIT(1) HGYSMIAMHSIH(1) TFQIPSTFVWHR(1) FHGRHGLEPLQV(1) TPTSALLPPLAY(1) APQDNTIMPSPT(1) SQPFFAWLKNIP(1) [FW], P, W, T 16 Phage display (subtractive panning) 3 PMID:19690339 M-CLL mAb255 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection on BSA-blocked Protein A and Protein G beads was carried out in rounds 2 and 3 before incubation with the CLL mAb. 2306 YHHAYLLPPPRM(2) TDSYYHHPFSKY(1) LPNIPYHHPLYH(1) YHHPIVGQVQFI(1) LPPSAYFHHAYL(1) YSWAHHTYPPLT(1) HPNYHHGYPSVS(1) SPKSEHIHHSFA(1) ESWSQYYHHSHP(1) YHHMPMIGRTII(1) ALTYPSAYHHGW(1) DVVPTYNYAFHH(1) QDRLPNRWHTYI(1) P, YHH, [FY], [ST] 13 Phage display (subtractive panning) 3 PMID:19690339 M-CLL mAb412 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection on BSA-blocked Protein A and Protein G beads was carried out in rounds 2 and 3 before incubation with the CLL mAb. 2307 IPLPPPSRPFFK(27) SLVPHPRFTNHN(1) WGSSARYVLPMP(1) HYPTAKFHAERL(1) P 4 Phage display (subtractive panning) 3 PMID:19690339 U-CLL mAb014 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection on BSA-blocked Protein A and Protein G beads was carried out in rounds 2 and 3 before incubation with the CLL mAb. 2308 WPLWSIPPFSPR(10) TQTTVSSTQKPY(1) SGASRTPAALSY(1) SWASLPVGMNSW(1) QLHMNMSSRTDT(1) EAPQKLDPRYAQ(1) [ST], [VL]-P 6 Phage display (subtractive panning) 3 PMID:19690339 U-CLL mAb068 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection on BSA-blocked Protein A and Protein G beads was carried out in rounds 2 and 3 before incubation with the CLL mAb. 2309 WHDDQMLRRTVT(10) DTPYWWIPHFNA(2) SHRLDTLLLLAT(1) YHHSYPPIYTGW(1) H 4 Phage display (subtractive panning) 3 PMID:19690339 U-CLL mAb114 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection on BSA-blocked Protein A and Protein G beads was carried out in rounds 2 and 3 before incubation with the CLL mAb. 2310 FSLQTMRHWPYS(16) WTPDDGWRTNLT(1) SSNYQMYYQYFS(1) TTDKHSMTPAAS(1) QLIPHIDAVRLL(1) ASLQGMQTSSWA(1) MNPPWKHTMPRQ(1) QYNMVYAPDVPL(1) SPNFSWLPLGTT(1) TMASSMWPLNRW(1) MPAVMSSAQVPR(1) WPMRLEASYAKP(1) NTKPSLPTISWP(1) WVEILPTARPHG(1) TWIPSAWLIDWD(1) 15 Phage display (subtractive panning) 3 PMID:19690339 U-CLL mAb270 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection on BSA-blocked Protein A and Protein G beads was carried out in rounds 2 and 3 before incubation with the CLL mAb. 2311 WNLRDTCYPYCT(13) WHDDQMLRRTVT(5) TVKVTDFANLQR(1) MPANSPTSPAFP(1) ATPGITHYTSSA(1) FARGGGELPESM(1) NSMPVRPLLQDF(1) ETYLVRETLLRQ(1) TTVAKRKRTQSR(1) RVKMSKNKKNPA(1) AGVHFTATKAAP(1) LDFSYSRTSWAP(1) FHDNETLRKRVT(1) W, [ST]-[HKR], R 13 Phage display (subtractive panning) 3 PMID:19690339 U-CLL mAb114 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection was performed in rounds 2 and 3 by incubating the amplified eluate of the previous round with bead slurry (50 L) of Sepharose-polyclonal human IgG. 2312 CGFSGELGC 1 Phage display (subtractive panning) 2 PMID:24098722 Atherosclerotic plaques Not determined. Not determined. Not determined. CX7C fUSE5 phage display library The phage library in 100 μL of DME with 3% BSA was incubated with the luminal face of the normal carotid. Unbound phages were recovered and incubated for 2 hours with the luminal face of a fresh carotid plaque. Comparison to public databases using BLASTp indicated that one of the phages (phage B6, and only this phage) presented four consecutive amino acids (ELGC) in nine (CGFSGELGC) identical to plexin B1, rendering an alignment score of 16.3. 2313 WSGPGVWGASVK(3/31) TLSGAFELSRDK(3/31) TPWQDTIKPQRE(3/31) 3 Phage display (in vivo) 2 PMID:24105738 Ovarian cancer Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) SKOV3 suspensions were subcutaneously injected into the right flank of mice to generate epithelial ovarian cancer xenografts. The phage-displayed peptide library (New England Biolabs, Beverly, MA, USA), e11 TU was intravenously injected and circulated for 15 min. After anesthetized with intra-peritoneal injection of chloroquine, the mice were perfused through the heart. Then, the tumor tissues were collected, homogenated with a micro tissue grinder, and then washed, weighed and lysed with 1% NP40. Other 22 phage clones in the second round appeared once respectively, but their sequences were not shown. 2314 WSGPGVWGASVK(39/43) TLSGAFELSRDK(1/43) VTPTEHFTGSWR(1/43) HTHTTTHWFTSN(1/43) YPSELNVRTPHR(1/43) 5 Phage display (in vivo) 3 PMID:24105738 Ovarian cancer Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) SKOV3 suspensions were subcutaneously injected into the right flank of mice to generate epithelial ovarian cancer xenografts. The phage-displayed peptide library (New England Biolabs, Beverly, MA, USA), e11 TU was intravenously injected and circulated for 15 min. After anesthetized with intra-peritoneal injection of chloroquine, the mice were perfused through the heart. Then, the tumor tissues were collected, homogenated with a micro tissue grinder, and then washed, weighed and lysed with 1% NP40. To evaluate the homing efficacy, phage clone PC3-1 (WSGPGVWGASVK) and M13KE control phage were injected into the xenografts model. As a comparison, M13KE phage showed no immunoactivity. Based on these results, it was obvious that the peptide WSGPGVWGASVK displayed on PC3-1 was a suitable candidate for ovarian cancer-targeting peptide. 2315 HLYVSPW 1 Phage display (common panning) 4 PMID:23852533 Toll-like receptor 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Five phage clones (E5, F7, Pep2, F11 and E6) were selected, but their sequences were not shown. The TLR2-binding CPP, HYLVSPW, is a promising candidate for targeted drug development. 2316 SNHWHSQGLVPR(6) ANLTPTGLVPRP(1) QLYDHLGLVPRL(3) GSPLTGLVPRWA(2) GLVPR 4 Phage display (common panning) 3 PMID:23420525 Anti-Efb-C polyclonal antibody Clumping factor A and B Not determined. Not determined. Ph.D.-12 phage display library (X12) 2317 STMWPSV(1/8) SVTNHPT(1/8) TDIQHAK(1/8) HLPVPLR(1/8) YPYPLIS(1/8) LHSELPT(1/8) SSLSLTP(1/8) RPMNIGS(1/8) 8 Phage display (subtractive panning) 3 PMID:23784990 Ceramic fluorapatite Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The original phage library was diluted in 1 ml TBST (0.1%) buffer and incubated with the blocked empty column (without CFA sample) at room temperature. After 1 h of incubation, the phage solution was discarded. The discarded solution contains the phages that did not show strong affinity to the column materials and was used for the selection of CFA-binding phages. 2318 HLPPWTQ(1/10) KPRSMLH(1/10) KLHLPLH(1/10) MVHAQRP(1/10) AYHPDAT(1/10) SQAHFYE(1/10) SKTKNNL(1/10) KTTGLTV(1/10) 8 Phage display (subtractive panning) 4 PMID:23784990 Ceramic fluorapatite Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The original phage library was diluted in 1 ml TBST (0.1%) buffer and incubated with the blocked empty column (without CFA sample) at room temperature. After 1 h of incubation, the phage solution was discarded. The discarded solution contains the phages that did not show strong affinity to the column materials and was used for the selection of CFA-binding phages. 2319 SQMHLRY(1/25) KGFHGYT(1/25) LLTKNSL(1/25) KPRSVSG(11/25) VMAPHKY(1/25) QRHTRPN(1/25) YLPPYKS(1/25) DHPAVAQ(1/25) SSPDKSF(1/25) HRPYIAP(1/25) ASEQVHM(1/25) SRLTMHT(1/25) HEVSDYL(1/25) THSKPLH(1/25) NKITQHS(1/25) 15 Phage display (subtractive panning) 5 PMID:23784990 Ceramic fluorapatite Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The original phage library was diluted in 1 ml TBST (0.1%) buffer and incubated with the blocked empty column (without CFA sample) at room temperature. After 1 h of incubation, the phage solution was discarded. The discarded solution contains the phages that did not show strong affinity to the column materials and was used for the selection of CFA-binding phages. The peptide F5-4 ((KPRSVSG)), displayed by more than 40% of the phages analyzed in the fifth round of selection, was subjected to further studies. Selectivity of the peptide for the chemical composition and morphology of CFA was assured by the adsorption studies. The dissociation constant, obtained from the F5-4/CFA adsorption isotherm, was in the micromolar range, and the maximum capacity was 39.4 nmol/mg. 2320 MQNPTQAPSVGL(20) QDPTNSGFYGRP(3) LPLRTDPWAKAS(2) 3 Phage display (common panning) 2 PMID:23740479 Palladium powder Not determined. Not determined. Not determined. Ph.D.-12 phage display and wild type library pool 2321 AGRSYW(16) QARVAE(15) RAVLEA(1) RLQSRT(2) KPTTGR(2) QLQSRL(1) RLQSRR(1) 7 Phage display (common panning) 5 PMID:24219291 Serine protease 29 Not determined. Not determined. Not determined. T7 X6 phage display library 2322 CMSYEGSWRKWVMWGGC CDLFVMAVGTNLDWWGC CVERCLASTSSGVKALC 3 Phage display (common panning) 5 PMID:23896727 Tumor necrosis factor receptor superfamily member 5 CD40 ligand, CD40-L 3QD6, Not determined. pComb8 CX15C phage display library Among 5 selected phage colonies, the most frequently observed peptide sequence was NP31 (CMSYEGSWRKWVMWGGC) (60%), while 20% of clones were identical to the NP26 (CDLFVMAVGTNLDWWGC) and NP58 (CDLFVMAVGTNLDWWGC) clone sequences. 2323 DGNVVTCWACREKRW 1 Phage display (common panning) 5 PMID:23896727 Tumor necrosis factor receptor superfamily member 5 CD40 ligand, CD40-L 3QD6, Not determined. pIF15 phage display library 2324 WHKQSQWPFGVW(10) AQIPSSLFYPPR(1) WAYPERAVFQFL(1) NTLNWKWFSPPF(1) SLAYPTPDARHL(1) SFTPPFPRLSLT(1) LLADTTHHRPWT(2) WHYNSQLIETLS(2) HQLPISISPHYP(1) WHYNSQADIDSF(1) GHKQSQWPFGVM(1) ETQSLPVQSMPP(1) WHQNWMYTAMQL(1) HHKTVWFSNFHS(5) TNPTLPA(1) EHPTSHINSTRP(1) TMGFTAPRFPHY(1) TLSNLSRVHAVL(1) HTMMVLPLGAKS(1) SLTVPFLPLYVP(1) TYTTAGSASRLV(1) HTRHITQAATTA(1) SQNVRSDPWKSQ(1) SADMHKRSGGPY(1) QFLDLHMRVTPV(1) NLTHPHRIPWFE(1) TMGFRAPRFPHY(1) SDNVHTWQAMFK(1) SAHGTSTGVPWP(1) KYPIQTSWLDSV(1) YQPMYSLLLRPG(1) TYTTAGQSSRLV(1) GNSWFIEQVVLL(1) TMGFTRPRFPHY(19) SQPFFAWLKNIP(1) NHLPGSPPETRS(1) HITLRMTDTESR(1) SVSVGMKPSPRP(1) 38 Phage display (common panning) 3 PMID:22171001 Replication initiator protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2325 AVEYYRFDS(1) DEVVFYLGF(1) DMVWLMYRL(1) ELVLFMGGW(1) EWMVWFTGV(1) GFSFFWGRW(1) GLMFFQGGW(1) GLTWFMGTW(1) IEWYSGRLW(1) IIWYVALPE(1) LLYFRGVEV(1) LMWFRGAFD(1) LMWFRGLSI(1) LVLFASRVV(1) LVMFSGTYW(1) LVWYNLPLS(2) LVYYVSRRM(1) MLTWFFSTP(1) RWMLVWFRR(1) SLVWFRSKF(1) VAGLGSFGH(1) VVWFRGVFS(1) VVWYSLVPY(1) VVWYVATLS(1) WIWYNSVLK(1) WLFFNGSLW(1) WLVWFRGQR(1) WVFFQSALP(1) WVFFSYSEL(1) YIWFLGEMP(1) YTYYSLLVV(1) LVWFRG 31 Phage display (common panning) 6 PMID:11896050 Mast cell protease 4, mMCP-4 Not determined. Not determined. Not determined. X9 T7 phage display library 2326 TLMVPRTGS(4) MASPRMLR(1) GAKPRALR(1) SRASRLKV(1) RWSPRSIYG(1) 5 Phage display (common panning) 6 PMID:11896050 Mast cell protease 4, mMCP-4 Not determined. Not determined. Not determined. X9 T7 phage display library 2327 YFRILRIKGLSS(2) SLSLITMLKISR(2) HMRTILPLKLTI(1) RPPIRNPININS(1) RPSYLSLIRILK(1) IHRYAEHRILPN(1) LRIIIKTRRLRL(1) RIQRIPILMINP(1) KPSRTIRILSHN(1) IPTIKSTKSIRN(1) RIYFIKWRRISL(1) QLRRRSMLITPH(1) RIRIHTPRLIIP(2) RMRTPIRHMILP(1) HHQRILPMRKIM(1) IKLLIKMNLKRN(1) LLKHRPRINKNL(1) RNKILTQRIINN(1) IPKIMHLLKRTM(1) RLHFILKWIIHR(1) RMRINQRNLMIN(1) AFPTRMRIQKTI(1) RPTLISLIYRMI(1) AYILPRIKSFTA(1) 24 Phage display (subtractive panning) 4 PMID:24014474 Hepatocellular carcinoma cell line HepG2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2328 CGX(F/Y/W)(S/N)HPQC 1 Phage display (common panning) 5 PMID:15919886 Streptavidin Biotin Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2329 IPLPPPSRPFFK 1 Phage display (subtractive panning) 4 PMID:22791264 Platelet-derived growth factor receptor β (PDGFRβ) Platelet-derived growth factor subunit B Not determined. Not determined. Ph.D.-12 phage display library (X12) Both positive (PDGFRβ) and negative (FGFR) target were biotinylated. Each selection round was conducted as follows: e11 plaque-forming units were added in 500μl PBST supplemented with 0.1% BSA pH 7.0. After 30-min incubation, 50μl Dynabeads? M-280 Streptavidin (Invitrogen) was added together with 100 μl biotinylated negative target FGFR 100 nM and 100μl non-biotinylated FGFR 1,000 nM. After 1-h incubation at room temperature, the unbound phages were separated with a Dynal magnet and incubated with 100 μl biotinylated PDGFRβfor 1 h at room temperature. Eighty percent of the clones isolated on biotinylated target in suspension displayed the peptide sequence: IPLPPPSRPFFK. 2330 EHMALTYPFRPP(16) AYYPQNHKSKAE(15) APNHIPRPPGLT(10) YPHYSLPGSSTL(8) AHRHPISFLSTL(2) SILSTMSPHGAT(1) ITMSSNAEHSRI(1) 7 Phage display (subtractive panning) 3 PMID:19327883 NCI-H1299 non-small cell lung cancer cell line Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) NCI-H1299 cells were taken as the target cells, and the normal lung cell line (SAEC) as the absorber cells for a whole-cell subtractive screening from a phage display 12-peptide library. 2331 GRVRDQVAGW GRVKDQIAQL GVRDQVSWAL ESVREQVMKY SVRSQISASL GVRETVYRHM GVREVIVMHML GRVRDQIWAAL AGVRDQILIWL GRVRDQIMLSL G-R-V-R-D-Q-I-x(3)-L 10 Phage display (common panning) 3-4 PMID:9180079 Thrombopoietin receptor, TPO-R Thrombopoietin Not determined. Not determined. pⅧ and peptides-on-plasmids phage display library pool 2332 CTLRQWLQGC CTLQEFLEGC CTRTEWLHGC CTLREWLHGGFC CTLREWVFAGLC CTLRQWLILLGMC CTLAEFLASGVEQC CSLQEFLSHGGYVC CTLREFLDPTTAVC CTLKEWLVSHEVWC C-T-L-R-E-W-L-x(2-6)-C 10 Phage display (common panning) 3-4 PMID:9180079 Thrombopoietin receptor, TPO-R Thrombopoietin Not determined. Not determined. pⅧ and peptides-on-plasmids phage display library pool 2333 REGPTLRQWM EGPTLRQWLA ERGPFWAKAC REGPRCVMWM CGTEGPTLSTWLDC CEQDGPTLLEWLKC CELVGPSLMSWLTC CLTGPFVTQWLYEC CRAGPTLLEWLTLC CADGPTLREWISFC C-x(1-2)-E-G-P-T-L-R-E-W-L-x(1-2)-C 10 Phage display (common panning) 3-4 PMID:9180079 Thrombopoietin receptor, TPO-R Thrombopoietin Not determined. Not determined. pⅧ and peptides-on-plasmids phage display library pool 2334 CAAERGLFEDC(4) CTAWTYVLGPC(1) CTSWAYVLGPC(7) 3 Phage display (common panning) 3 PMID:11520027 Anti-E-3-G monoclonal antibody 41551 Estrone-3-glucuronide, E-3-G Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) 2335 CERGPGKSRSCS CGNRVSKAPKS GRKVKCS RAAMEKPS RRAAVRMEKPCS 5 Phage display (subtractive panning) 1 PMID:11820288 Anti-MUC1 monoclonal antibody C595 Mucin-1, MUC-1 Not determined. Not determined. T7 CX9C phage display library The library was first screened against an 'irrelevant 'antibody that would not be expected to bind specifically to MUC1 epitopes. Anti-Estrone beta-D-glucuronide antibody was used for this purpose. 2336 CSRVAPNRK CVKRTASGSGCS CSMRASGGPKCS CTVPVRPQQKCS CPATTHLG CHLAGT CEE C 8 Phage display (subtractive panning) 2 PMID:11820288 Anti-MUC1 monoclonal antibody C595 Mucin-1, MUC-1 Not determined. Not determined. T7 CX9C phage display library The library was first screened against an 'irrelevant 'antibody that would not be expected to bind specifically to MUC1 epitopes. Anti-Estrone beta-D-glucuronide antibody was used for this purpose. 2337 RNREAPRGKICS(7) RNREAPRGKIC(1) RRPPMTTASCS(1) FERIAPKGGNCS(1) CSRGPAGRTVCS(1) CRAPAGSKKMCS(1) 6 Phage display (subtractive panning) 3 PMID:11820288 Anti-MUC1 monoclonal antibody C595 Mucin-1, MUC-1 Not determined. Not determined. T7 CX9C phage display library The library was first screened against an 'irrelevant 'antibody that would not be expected to bind specifically to MUC1 epitopes. Anti-Estrone beta-D-glucuronide antibody was used for this purpose. 2338 RNREAPRGKICS(4) NREAPRGKICS(1) CRPAPSAKVACS(1) FERIAPKGGNCS(1) CDSERTAPKCS(1) RQAGRKPVNNCS(1) CSRGPAGRTVCS(1) CSDRMPCEPSCS(1) RRPSR(1) 9 Phage display (subtractive panning) 4 PMID:11820288 Anti-MUC1 monoclonal antibody C595 Mucin-1, MUC-1 Not determined. Not determined. T7 CX9C phage display library The library was first screened against an 'irrelevant 'antibody that would not be expected to bind specifically to MUC1 epitopes. Anti-Estrone beta-D-glucuronide antibody was used for this purpose. 2339 WHWTWLSEYPPP LETSKLPPPAFL WHWRNPDFWYLK 3 Phage display (common panning) PMID:19813732 Gamma-gliadin and alpha/beta-gliadin Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2340 CATLDGVSC CFAGAGVRC CGGRHWVRC CGSVLPVLC CGSVSHRRC CIGGRWVVC CNSVRGSRC CRRHSVSGC CRSGRVSNC CRVWHRGGC CSAAGLARC CSGAFWGSC CSGWFAGSC CSPSSGTYC CVVWRGGIC 15 Phage display (subtractive panning) 2 PMID:16397212 Epidermal growth factor receptor Pro-epidermal growth factor, EGF 1IVO,1NQL,3NJP, Not determined. Ph.D.-C7C phage display library (CX7C) To isolate peptides binding to epidermal growth factor receptor (EGFR), phage clones selected on SKOV3 in rounds 2 and 3 of the screening were individually amplified and pooled, and e9 transduction units of the mixed phage were incubated overnight with 10 μg of purified human EGFR. 2341 LWCITQDCSHRM(4) HDAISWTHYHPW(1) 2 Phage display (common panning) 5 PMID:24111863 Aminopeptidase N Transmissible gastroenteritis virus, TGEV Not determined. Not determined. Ph.D.-12 phage display library (X12) MTT assay results indicated that the PM1 (HDAISWTHYHPW) phage clone had a protective effect against TGEV infection in ST cells, whereas the PM1 and PM4 (LWCITQDCSHRM) phage clones had no effect on swine testis (ST) cell growth. 2342 SVSVGMKPSPRP(4) LPWHFKSRHRYQ(1) EWMSHGHPRPNN(1) SLSTPATRHFSG(1) LSTPYSKSQAST(1) SHWNSHSTPARA(1) ALSTPTFSTLPA(1) 7 Phage display (common panning) 3 PMID:24130862 Anti-rTs-Pmy-N monoclonal antibody 8F12 Paramyosin, Ts-Pmy Not determined. Not determined. Ph.D.-12 phage display library (X12) Mice immunized with KLH-conjugated peptides 8A1 (ALSTPTFSTLPA), 8F1 (LPWHFKSRHRYQ) and 8F7 (LSTPYSKSQAST) and subsequently challenged with T. spiralis larvae resulted in a 22.7%, 22.2% and 26.2% reduction in muscle larvae burden, respectively. However mice immunized with th KLH-conjugated peptide 8F6 (SHWNSHSTPARA) had only 18.8% reduction in muscle larvae burden compared to the KLH control (p< 0.05). 2343 LSPPRYP(5) 1 Phage display (subtractive panning, competitive panning) 3 PMID:24142482 FGFR-expressing Balb/c 3T3 cells Fibroblast growth factors Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage library was first incubated with FGFR-deficient Cos-7 cells. Then the supernatant was collected and added to the flask cultured with Balb/c 3T3 cells. For the last round of selection, low affinity cell-bound phages were first eluted with bFGF for 1 h, discarded, and high-affinity cell-bound phages were then eluted with bFGF for an additional 1 h. The synthetic P9 (LSPPRYP) peptide could inhibit the activation of two MAPKs (Erk1 and Erk2) in B16-Fl0 cells after stimulation by bFGF. And also the P9 peptide has a potent inhibitory effect on tumor growth in vivo, which was associated with significant inhibition of Erk1/Erk2 activation in tumor cells. 2344 FSDHWVN(6) 1 Phage display (common panning) 3 PMID:24145794 Anti-SAH monoclonal antibody ab111903 S-adenosyl-L-homocysteine, SAH Not determined. Not determined. Ph.D.-7 phage display library (X7) The original phage library was first incubated with anti-SAH Mab.The antibody-bound phages were then captured with Protein A or Protein G magnetic beads. Protein G magnetic beads were used in the first and third rounds. Protein A magnetic beads were used in the second round. To determine the affinity of the selected phage from the third-round, eight phage clones isolated and amplified. Six of these clones showed significant binding to the SAH antibody which was competitively inhibited by SAH, whereas two showed no significant binding. This strongly suggested that these six phage clones bound the antigen binding site of the SAH antibody. The six binding phage clones had the peptide sequence FSDHWVN. 2345 CRGATPMSC(1) CSEGLLNTC(1) CVQMPAHSC(1) CPNSTHRNC(1) CMHTHSRMC(1) CNTGSPYEC(1) CFSGMSTDC(1) CDASRPATC(1) 8 Phage display (common panning) 3 PMID:24145794 Anti-SAH monoclonal antibody ab111903 S-adenosyl-L-homocysteine, SAH Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The original phage library was first incubated with anti-SAH Mab.The antibody-bound phages were then captured with Protein A or Protein G magnetic beads. Protein G magnetic beads were used in the first and third rounds. Protein A magnetic beads were used in the second round. 2346 LRFLCWLDPDWMDCA(1) HGLSAWYLWALDAVG(1) ISGWYWWPIVDLYGA(1) QAMISPSRGFPRKYS(1) GSSFFYDVCSMWSLC(1) SLVYRSLFLDCHSDR(1) PFVDILVGLGLYPAG(1) DLTDFWNAFVPRIFG(1) VWLGFVDALHAFSGP(1) FLCMFAAHVAPYYCC(1) AYWFKCLALAYPGVA(1) FWGALLPPLLLTMGA(1) DPRASHLTGIRQLLQ(1) CFYDVASAIWASLSA(1) IFCVPSWLFSALYPD(1) DFWWSLLGDVLMVPQ(1) WFGLAADVGSKWLTW(1) VFPSPWAFLEPWWNT(1) ACFVDPSCWWSLLAR(1) FYLWSMLSALDAFTL(1) LLRYIPWLVPLPIGN(1) IFYSYLNAALGFPSI(1) VFDSLL 22 Phage display (common panning) 4 PMID:24251365 MA026 Not determined. Not determined. Not determined. X15 T7 phage display library Twenty seven single phage clones from the fourth-round elution were randomly picked out. Multiple sequence alignment analyses using CLUSTALW indicated that peptides 1 (FLPFWYNVFDSLSGN), 3 (FSAWLYSVFDQFSPA), 10 (GSSFFYDVCSMWSLC), 19 (CFYDVASAIWASLSA), and 24 (ACFVDPSCWWSLLAR) shared homology. Among them, the peptide sequence VFDSLL, a partially homologous sequence, was found. Then a single protein that includes the VFDSLL sequence, claudin-1 (CLDN1), was identified in the NCBI database by using BLAST. CLDN1 is highly expressed in the liver and plays an important role during the post cell binding process of HCV entry. 2347 CFDTRSLVC(3) CDHGYLPSC(2) CHYDGARAC(1) 3 Phage display (subtractive panning) 4 PMID:24265677 Mycobacterium Dleu/Dpan strain Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Three positive rounds of panning were performed against the targeted M. tb Dleu/Dpan strain. In order to remove peptides binding to cell wall components common to the mycobacterium genus, a subtraction round (round 4) was performed against M. smegmatis. A final positive panning round (round 5) was performed to enrich for peptides that are specific to M. tb. Ten plaques were selected using the traditional random cloning picking from the final round of biopanning, and were sequenced. Sequencing data of four of these plaques were ambiguous. Three unique sequences were obtained from the six remaining randomly selected plaques. 2348 CTYYWPLPC CSWYWPLPC CLMTSQFRC CWPIKVGWC 4 Phage display (common panning) 4 PMID:24265799 Ephrin type-A receptor 4 Type A/B ephrins Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) After four rounds of selection on EphA4, we detected an approximately 75-fold enrichment of phage binding to EphA4. In total, 48 of the individual clones that were tested for the binding to EphA4 Fc, and 6 of them showed significant binding when compared with the BSA negative controls. All 6 of these EphA4-binding clones displayed four different cyclic peptide sequences. The cyclic nonapeptide, TYY, CTYYWPLPC, binds to EphA4, but not to EphB4, and selectively inhibits the binding of ligand ephrinA5 to EphA4. The TYY cyclic peptide was evaluated for its cytotoxic effects on HUVECs, and it effectively inhibits HUVEC tubule formation activity. 2349 LSNNNLR 1 Phage display (common panning) 5 PMID:24284895 Silicon dioxide, SiO2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2350 GPIPGLLATVAV(2) LDTHASHACSTG(1) MATQPKLVGSPY(1) SNSPTFVCHRMV(1) SVVTSHQRYGTS(1) VQFPIEAMFWST(1) 6 Phage display (common panning) 2 PMID:24287902 Quinoprotein glucose dehydrogenase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2351 LGDSSNSQVSLN(1) LPHSAVMAQLTY(1) STSHHHHPSAPS(1) TLHAHQHHQPST(1) WMNGPVSIRTWS(1) WPYNHHHRTPSP(1) YSHHHMHTPHTR(1) 7 Phage display (subtractive panning) 2 PMID:24287902 Quinoprotein glucose dehydrogenase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage display peptide library was incubated with Ni-agarose magnetic beads in microtubes to eliminate phages binding to the beads or the tubes. In the first round, 120 μg of GB-His in immobilization buffer was added to 120 μ L of Ni-agarose magnetic.The library was subsequently added to 120 μg of GB-His immobilized Ni-agarose magnetic beads. In the second round, the first-round-eluted phages were incubated with 48 μg of GB-His. In the third round (negative-selection round), the phages were incubated with DEE-His. 2352 APLSQHHHHLRP(1) ELITNSETTQWF(1) MHDLTAALSLPP(1) TMQPGQNSHPIL(1) TSHIHTTPHSHH(1) YHPANHSFQHHF(1) YNHHGHHLDKHR(1) 7 Phage display (subtractive panning) 2 PMID:24287902 Quinoprotein glucose dehydrogenase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage display peptide library was incubated with Ni-agarose magnetic beads in microtubes to eliminate phages binding to the beads or the tubes. In the first round, 120 μg of GB-His in immobilization buffer was added to 120 μ L of Ni-agarose magnetic.The library was subsequently added to 120 μg of GB-His immobilized Ni-agarose magnetic beads. In the second round, the first-round-eluted phages were incubated with 48 μg of GB-His. In the third round (negative-selection round), the phages were incubated with HI. 2353 SDLSPIQSLSAI(2) APLFTQTWGPWR(1) APYAHHHHPVTP(1) EELWHHHPPSHH(1) LGDSSNSQVSLN(1) NSTHHHHFATIW(1) QPHKYPHSHHGP(1) TSLHQHHPTAAF(1) VAQHSHHHVTPS(1) 9 Phage display (subtractive panning) 3 PMID:24287902 Quinoprotein glucose dehydrogenase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage display peptide library was incubated with Ni-agarose magnetic beads in microtubes to eliminate phages binding to the beads or the tubes. In the first round, 120 μg of GB-His in immobilization buffer was added to 120 μ L of Ni-agarose magnetic.The library was subsequently added to 120 μg of GB-His immobilized Ni-agarose magnetic beads. In the second round, the first-round-eluted phages were incubated with 48 μg of GB-His. In the third round (negative-selection round), the phages were incubated with DEE-His. In the forth round, the third-round-eluted phages were screened against 2-μg wild-type GDH-B immobilized in a well of a 96-well ELISA plate. 2354 SPHSHHMSPSEY(2) LSPHHHHLDGHI(2) ANPLHHHHLWEL(1) NSTHHHHFATIW(1) QPHKQAVSFAFA(1) QTSHYHHHRAHT(1) TPTFPYWYGSLT(1) VHTHHLGHQPVR(1) 8 Phage display (subtractive panning) 3 PMID:24287902 Quinoprotein glucose dehydrogenase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage display peptide library was incubated with Ni-agarose magnetic beads in microtubes to eliminate phages binding to the beads or the tubes. In the first round, 120 μg of GB-His in immobilization buffer was added to 120 μ L of Ni-agarose magnetic.The library was subsequently added to 120 μg of GB-His immobilized Ni-agarose magnetic beads. In the second round, the first-round-eluted phages were incubated with 48 μg of GB-His. In the third round (negative-selection round), the phages were incubated with HI. In the forth round, the third-round-eluted phages were screened against 2-μg wild-type GDH-B immobilized in a well of a 96-well ELISA plate. 2355 CDRRDLPDWAIRAC 1 Phage display (common panning) 4 PMID:24304511 HIV-1 CDC451 gp120 Not determined. Not determined. Not determined. CX12C phage display library The peptide, CDRRDLPDWAIRAC, not only bound to gp120 but also enabled the binding of the stringent CD4i mAb CG10. 2356 LYQDYSL(2) EPLQLKM(2) LPWKPLG(2) TPAHPNY(2) LGAQSNF(1) PGAQSNF(1) VNSPTHS(1) VNSATHS(1) YQDSAKT(1) 9 Phage display (in vivo) 3-4 PMID:24320790 Four- to five-week-old mdx mice heart Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Four- to five-week-old mdx mice were injected intravenously with phages (one animal per round). Mice were perfused with phosphate-buffered saline (PBS) after 1 hour to remove unbound phages. The gastrocnemius and heart muscles were isolated and washed three times with Tris-buffered saline (TBS). Subsequently the organs were homogenized in 2 mL TBS with MagNA Lyser green beads (Roche Diagnostics) in the MagNA Lyser (Roche Diagnostics). The suspension was titrated and amplified. Up to four rounds of biopanning were performed for each fraction. After the third and fourth round, phages were plated and colonies were picked. The selected peptides were synthesized with a FAM label and incubated with primary human control myotube cultures and analyzed with fluorescence microscopy. Clear fluorescence was observed for 7 of the 12 peptides. Based on the highest fluorescence intensity, P1 (LYQDYSL), P2 (LPWKPLG) and P4 (LGAQSNF) were chosen for in vivo evaluation. Mice were intramuscularly injected with labeled peptide in the gastrocnemius muscle. Analyzed with fluorescence microscopy, clear fluorescence was observed for P4 at the membrane. P4 was conjugated to a 2OMePS AON specific for mouse dystrophin exon 23 (23AON). Upon systemic administration in dystrophic mdx mice, conjugation of a 2'-O-methyl phosphorothioate AON to this peptide indeed improved uptake in skeletal and cardiac muscle, and resulted in higher exon skipping levels with a significant difference in heart and diaphragm. 2357 EPLQLKM(2) TALPPSY(2) AMISAIH(1) HVIANAG(1) GNTPSRA(1) 5 Phage display (in vivo) 3-4 PMID:24320790 Four- to five-week-old mdx mice muscle Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Four- to five-week-old mdx mice were injected intravenously with phages (one animal per round). Mice were perfused with phosphate-buffered saline (PBS) after 1 hour to remove unbound phages. The gastrocnemius and heart muscles were isolated and washed three times with Tris-buffered saline (TBS). Subsequently the organs were homogenized in 2 mL TBS with MagNA Lyser green beads (Roche Diagnostics) in the MagNA Lyser (Roche Diagnostics). The suspension was titrated and amplified. Up to four rounds of biopanning were performed for each fraction. After the third and fourth round, phages were plated and colonies were picked. The selected peptides were synthesized with a FAM label and incubated with primary human control myotube cultures and analyzed with fluorescence microscopy. Clear fluorescence was observed for 7 of the 12 peptides. Based on the highest fluorescence intensity, P11 (HVIANAG) were chosen for in vivo evaluation. Mice were intramuscularly injected with 2.5 nmol of labeled peptide in the gastrocnemius muscle. Analyzed with fluorescence microscopy, P11 was found negative. 2358 DSSPYLMSPLGLDFD SPDDPPLPDLLYRSG FAPDLTRFPSVVVST SSADGGQGPHLLVRY EHFAPSSPDFLERHF NVSPDALEWLVGSKC DRRFFQSDILALFSP LPPPQQFHQDMMKLF RLPLDTFHSDLSRLT DTADLWWNSGTFLPA THASYMSPSSAFTLQ 11 Phage display (subtractive panning) 2 PMID:24361045 Mixtures of 13 anti-dystrophin monoclonal antibodies Dystrophin Not determined. Not determined. f88-15mer phage display library (X15) A sample of the phage library was pre-incubated in dishes coated with the rabbit anti-mouse antibodies alone to ensure any binding was specific for the target mAbs. Then unbound phages were incubated with monoclonal antibody mixtures. 2359 KQPAFNL(5) KAPAFDL(4) KLIAFDL(4) KLPAFML(2) KAPAFNL(1) KSPAFNL(1) 6 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of the peanut allergic patient no. 2205 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgE of the peanut allergic patient no. 2205 were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (intact Ara h 1). Three rounds of positive selection were performed. 2360 KAPAFDL(3) KSPAFDL(3) KLPAFML(2) KQPAFNL(2) KTAAFNL(2) KAPAFNL(1) KEPAFML(1) KLPAFQL(1) 8 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of the peanut allergic patient no. 2205 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgE of the peanut allergic patient no. 2205 were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (digested Ara h 1). Three rounds of positive selection were performed. 2361 ASPKSLL(1) FHTARPW(1) FSPIVLY(1) SPIMDYF(1) SPPRGIF(1) TSRADTL(1) TTSVRNT(1) VTLAPLR(1) S-P-I-x(2)-[HY]-Y 8 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of the peanut allergic patient no. 2208 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgE of the peanut allergic patient no. 2208 were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (intact Ara h 1). Three rounds of positive selection were performed. 2362 SPIISHY(2) SPILAHY(2) MSDRGIF(2) AKPASWA(1) DSPRGVF(1) SPIINYY(1) SPITLYY(1) VPPRGLF(1) YDVSSLP(1) S-P-I-x(2)-[HY]-Y 9 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of the peanut allergic patient no. 2208 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgE of the peanut allergic patient no. 2208 were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (digested Ara h 1). Three rounds of positive selection were performed. 2363 ATETSYR(1) HAPRGVF(1) LPPRGLF(1) MSDRGIF(1) SIPYPAP(1) SNATWVP(1) SPIINYY(1) STIMSSR(1) WSPIVHP(1) P-P-R-G-[LV]-F 9 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of the peanut allergic patient no. 2209 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgE of the peanut allergic patient no. 2209 were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (intact Ara h 1). Three rounds of positive selection were performed. 2364 QWAFAIP(2) DLTWAPK(1) FSPIIAF(1) GLAGYPP(1) LSPIIVY(1) MSDRGIF(1) NLVLFSY(1) SPILAHY(1) SPIVLYF(1) SPIVNNY(1) S-P-I-x(2)-[HY]-Y 10 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of the peanut allergic patient no. 2209 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgE of the peanut allergic patient no. 2209 were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (digested Ara h 1). Three rounds of positive selection were performed. 2365 IPYTLDK(3) TPYTLDK(2) AKQTDTM(1) DISRGLF(1) EWSRGIF(1) LHRPLHP(1) VTGPAKT(1) 7 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of the peanut allergic patient no. 2304 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgE of the peanut allergic patient no. 2304 were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (intact Ara h 1). Three rounds of positive selection were performed. 2366 LPYTLSK(3) AKQTDTM(2) ADMRGLF(1) AKGTDNW(1) GSAFSAF(1) GVYSLSK(1) IPYTLDK(1) SFPARFY(1) TPYTLDK(1) WPDRGIF(1) 10 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of the peanut allergic patient no. 2304 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgE of the peanut allergic patient no. 2304 were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (digested Ara h 1). Three rounds of positive selection were performed. 2367 FSPIVLY(2) SPITEFY(2) KSIYHQW(1) LPSRGLF(1) LTDLTQK(1) QSLTRLP(1) SPILAHY(1) TFKLHPI(1) TPPRVHL(1) VLPGSKS(1) WESRGVF(1) YPPKPLH(1) S-P-I-x(2)-[HY]-Y 12 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of the peanut allergic patient no. 2305 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgE of the peanut allergic patient no. 2305 were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (intact Ara h 1). Three rounds of positive selection were performed. 2368 VMPGSKP(3) KNVYHQT(2) ACCRSIP(1) AVESNDK(1) FPPKPKL(1) LINQALK(1) MSDRGIF(1) SPIISHY(1) SPITLYY(1) SPITTYY(1) TPFSNSP(1) S-P-I-x(2)-[HY]-Y 11 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of the peanut allergic patient no. 2305 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgE of the peanut allergic patient no. 2305 were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (digested Ara h 1). Three rounds of positive selection were performed. 2369 AQSQFNS(5) YSTQVRP(2) AYTLPSR(1) HFSKVPR(1) SNAARAY(1) YHPFLQV(1) 6 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of animal nr. 19 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with mouse-α-rat IgE. For positive selection, IgE of animal nr. 19, rat immunized with digested Ara h 1, were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (intact Ara h 1). Three rounds of positive selection were performed. 2370 AQSQFNS(2) APHRHPH(1) APKAAPL(1) AQLAPET(1) GGQFGPP(1) IHLPPAL(1) NQTRTTH(1) QPMINML(1) QPPTANA(1) TPMAWSQ(1) VHNTRLL(1) 11 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of animal nr. 19 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with mouse-α-rat IgE. For positive selection, IgE of animal nr. 19, rat immunized with digested Ara h 1, were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (digested Ara h 1). Three rounds of positive selection were performed. 2371 AYPLRAH(1) GLTPSKN(1) GPPPLPK(1) HHLHQTN(1) LPLPVVR(1) NLANKMA(1) SLYKPHP(1) SSYSQLN(1) VTAPFRV(1) YGTHKSP(1) YSIPKSS(1) 11 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of animal nr. 49 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with mouse-α-rat IgE. For positive selection, IgE of animal nr. 49, rat immunized with intact Ara h 1, were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (intact Ara h 1). Three rounds of positive selection were performed. 2372 TASGLYS(4) GPPPLPK(1) HHMHTTR(1) IAMKPLA(1) LPSLPRI(1) LQSKTLH(1) NTSNPYT(1) QATFSHS(1) STFLPHP(1) VPPQLSR(1) VTAPFRV(1) VYPMAMS(1) YHGSVSL(1) 13 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of animal nr. 49 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with mouse-α-rat IgE. For positive selection, IgE of animal nr. 49, rat immunized with intact Ara h 1, were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (digested Ara h 1). Three rounds of positive selection were performed. 2373 STFLPHP(7) AMPPLPP(1) HPWAPMQ(1) KPPNLPN(1) KWLPTPL(1) SASWQES(1) SLTSWAT(1) TQMSKHL(1) VIAKTRL(1) 9 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of animal nr. 52 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with mouse-α-rat IgE. For positive selection, IgE of animal nr. 52, rat immunized with intact Ara h 1, were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (intact Ara h 1). Three rounds of positive selection were performed. 2374 AQSQFNS(4) ALGASHG(1) FHDTPQS(1) HISLGRI(1) LGTYGHY(1) NPASSHM(1) NYLSLLH(1) QIPSGTP(1) QLSAPPP(1) SASLMLQ(1) TMSKDST(1) YPPMTQV(1) YTQGWNL(1) 13 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of animal nr. 53 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with mouse-α-rat IgE. For positive selection, IgE of animal nr. 53, rat immunized with intact Ara h 1, were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (intact Ara h 1). Three rounds of positive selection were performed. 2375 AQSQFNS(3) AKTSSNV(1) APLPKLM(1) DQWHRAP(1) FGFPTTS(1) FPTPRVA(1) HQPQQLF(1) ISVNIQA(1) IVQHTVP(1) KPWLPQH(1) NAHHTYP(1) NHTTLKA(1) NPLPSQL(1) QPVPPRL(1) SDRFPPK(1) SDYHRVM(1) SLDTCLR(1) SPSPQRS(1) TGGVWSK(1) YNPAVIA(1) 20 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of animal nr. 53 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with mouse-α-rat IgE. For positive selection, IgE of animal nr. 53, rat immunized with intact Ara h 1, were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (digested Ara h 1). Three rounds of positive selection were performed. 2376 VTAPFRV(3) VIAKTRL(2) VPFKPIR(2) GMTMATP(2) GVGVPQR(2) AETVESC(1) DAFTRGT(1) GPPPLPK(1) IPSLPMR(1) LPVPIGY(1) MHTQDLM(1) MLNATSK(1) QDWNFKK(1) SGEIHFK(1) STFLPHP(1) YPTRELS(1) 16 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of animal nr. 54 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with mouse-α-rat IgE. For positive selection, IgE of animal nr. 54, rat immunized with intact Ara h 1, were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (intact Ara h 1). Three rounds of positive selection were performed. 2377 FTDTSWM(1) HGWPVPK(1) ITLQRTF(1) LHGRYFP(1) LTSPLRL(1) QGSQYTQ(1) TGSSNLY(1) TLLTTSP(1) TMTTPQQ(1) TTAPGKP(1) TTHYLHA(1) WPELYPV(1) YSYPGLT(1) 13 Phage display (subtractive panning, competitive panning) 3 PMID:22555070 IgE of animal nr. 54 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with mouse-α-rat IgE. For positive selection, IgE of animal nr. 54, rat immunized with intact Ara h 1, were used and phages were eluted by competitive immunoscreening, by adding 25 μg allergen (digested Ara h 1). Three rounds of positive selection were performed. 2378 GHTHKAM(1) GTSQFQL(1) HPPLLAT(1) HSMPTLH(1) HSSNAQV(1) HVPTVLS(1) IQQLKPP(1) ITPAALS(1) LNLDRKP(1) LPNPRNL(1) LQKTFMV(1) LSAKITL(1) LSTATVR(1) LTNSEWD(1) LTQQPPR(1) LWATRHI(1) LYALVPS(1) NQLLGST(1) QKPVYTR(1) QSFGVQL(1) SFPATKT(1) SHDVTPQ(1) STPLIPT(1) TPALFPY(1) TSPPRYL(1) VIHVKSP(1) VSHTQAN(1) YPRTISA(1) YSNAPTH(1) YSPHPYP(1) 30 Phage display (subtractive panning, competitive panning) 3 PMID:24365751 IgE of the peanut allergic patient no. 2208 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgG4 of the peanut allergic patient no. 2208 were used and phages were eluted by competitive immunoscreening, by adding phages were eluted by competitive immunoscreening by adding allergen (intact and digested Ara h 1) per 1 mL of beads. Three rounds of positive selection were performed. 2379 DLLSPAR(1) ISYQWPS(1) KVWVLPI(1) KVWYITS(1) LPARTVT(1) QYPRLTT(1) SHQIGPS(1) SLQEPWH(1) SPSFTQT(1) TDPTFAH(1) TLAQLPK(1) TRPLLPS(1) TVSPMTR(1) YHVPSYE(1) 14 Phage display (subtractive panning, competitive panning) 3 PMID:24365751 IgE of the peanut allergic patient no. 2209 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgG4 of the peanut allergic patient no. 2209 were used and phages were eluted by competitive immunoscreening, by adding phages were eluted by competitive immunoscreening by adding allergen (intact and digested Ara h 1) per 1 mL of beads. Three rounds of positive selection were performed. 2380 KCCYVPL(6) ANWTPYL(1) ESLQRPL(1) FAVQEAS(1) FHPTRYP(1) GLRWIAP(1) HITVPTR(1) HTNFTAP(1) HWHTSSR(1) KFTLHTP(1) KLWVIPV(1) LPLRLIL(1) LPSPWPY(1) LSLELAL(1) LTPYSSV(1) MATSTLA(1) MNYVPAP(1) MQPHYWQ(1) NHTYYQP(1) QTPESRS(1) SATMGKA(1) SFTTWLP(1) SLQSWLT(1) SMGIPRF(1) SPTLINL(1) TAPLHDP(1) TPLRTWP(1) TSTATSH(1) VPAHHLS(1) VSTTHIP(1) WSMYSAP(1) 31 Phage display (subtractive panning, competitive panning) 3 PMID:24365751 IgE of the peanut allergic patient no. 2305 Allergen Ara h 1, clone P41B Not determined. Not determined. Ph.D.-7 phage display library (X7) In negative selection, the library was incubated with rabbit-α-human IgE. For positive selection, IgG4 of the peanut allergic patient no. 2305 were used and phages were eluted by competitive immunoscreening, by adding phages were eluted by competitive immunoscreening by adding allergen (intact and digested Ara h 1) per 1 mL of beads. Three rounds of positive selection were performed. 2381 SVHLYHSTKTLR(4/11) QSYMERMYDAWP(4/11) DYMSALFMAHQT(3/11) 3 Phage display (common panning) 5 PMID:24460939 Interferon-induced, double-stranded RNA-activated protein kinase Eukaryotic translation initiation factor 2 subunit 1 Not determined. Not determined. Ph.D.-12 phage display library (X12) More than 100 clones were tested for their association towards PKRcat by phage-ELISA, and 11 clones with high binding index were subjected to DNA sequencing. Three sequences of high occurrence were obtained. By competitive phage-ELISA, the detected amount of phage was reduced with increasing concentration of P1 (DYMSALFMAHQT), P2 (SVHLYHSTKTLR), and P3 (QSYMERMYDAWP). Among the three peptides, P1 exhibited the most efficient binding with PKRcat. Besides, P1 and P2 inhibited PKR activity towards eIF2α in a concentration-dependent manner in vitro. P1 and P2 inhibit the enzymatic activity of PKR, and P1 is more efficient than P2 in vivo assay. 2382 LVFGTLLGQLRA(5) VSPSYYSWWNFR(3) F(3) NVSFR(2) CFMRL(1) CL(1) FAQMVIATNLSEM(1) FICIFRSSSVCG(1) IFVVLHFVCVHA(1) LPV(1) LSDHATFWASKV(1) PPSYLVLTGDSS(1) SVQYV(1) 13 Phage display (common panning) 6 PMID:21559518 Cyclosporin A, CsA Peptidyl-prolyl cis-trans isomerase A, PPIase A Not determined. Not determined. X12 T7 phage display library Due to the structural complexity of CsA, photoaffinity coupling method was applied to immobilize CsA. The highly reactive carbene induced by UV irradiation reacted with CsA, resulting in the production of immobilized CsA on solid surface in a nonspecific manner. Twenty two single phage clones were randomly pick out from the sixth panning elution. In order to validate the binding specificity of the phage, we amplified the phage #13 (LVFGTLLGQLRA) and measure the ratio of eluted phage titer with CsA and mock resins. The ratio of the phage #13 was 3.75, whereas randomly picked up phage was 1.00. These results indicated that the phage #13 specifically associated with CsA-immobilized resins. 2383 CDPRAADPC(1) CFGAPVSLC(1) CFPHLSRYC(1) CIWSPFFDC(1) CLYHDTHYC(1) CNPFCGSRC(1) CNPFLADPC(1) CPQDPKQWC(1) CPRSSLPSC(1) CPRSTSHLC(1) CPSPSRFPC(1) CQSTSGSSC(1) CSPDSLFPC(1) CSPWPLSYC(1) CTFSPLSVC(1) CTSPPLHAC(1) CVHSPFWPC(1) CYPGMLWTC(1) 18 Phage display (subtractive panning) 3 PMID:24256622 Serum of dogs with VL Leishmania infantum Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In negative selection, the library was incubated with IgGs that had been purified from healthy dogs. The supernatant containing the clones that were not adhered to the IgGs was recovered and transferred to a new tube, and this procedure using IgGs from healthy dogs was repeated three times. After this, the supernatant was recovered and transferred to a new tube containing microspheres coupled to IgGs from T. cruzi-infected dogs, and this procedure of negative selection was also repeated three times. For positive selection, phage clones that did not adhere to the IgGs from T. cruzi-infected dogs were incubated with IgGs purified from serum samples of dogs with asymptomatic VL. The bound phage clones were transferred to a new tube containing IgGs that had been purified from serum samples of dogs with symptomatic VL. The process was repeated three times with serum samples of dogs with asymptomatic and symptomatic VL. Approximately 96 clones were randomly selected from individual colonies, and their DNA sequences were PCR amplified and sequenced. Eighteen clones had their sequences clearly identified, and an alignment showed that no identical consensus motif could be detected between them. 2384 WNNWQPQ(4) 1 Phage display (competitive panning) 3 PMID:24096567 Anti-BDE47 monoclonal antibody 1H2 2,2′,4,4′-tetrabromodiphenyl ether, BDE47 Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage library was incubated with Mab 1H2 (20 μg/mL). Bound phages were eluted by incubation with PBS containing 100 ng/mL BDE47. The panning procedures were then repeated twice, but the Mab concentrations of 10 and 5 μg/mL were used for the second and the third panning, respectively. 2385 CWNNWQPQC(1) 1 Phage display (competitive panning) 3 PMID:24096567 Anti-BDE47 monoclonal antibody 1H2 2,2′,4,4′-tetrabromodiphenyl ether, BDE47 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage library was incubated with Mab 1H2 (20 μg/mL). Bound phages were eluted by incubation with PBS containing 100 ng/mL BDE47. The panning procedures were then repeated twice, but the Mab concentrations of 10 and 5 μg/mL were used for the second and the third panning, respectively. 2386 GVSRLEILRAQL(4) GVSRKEILRAQG(2) 2 Phage display (competitive panning) 3 PMID:24096567 Anti-BDE47 monoclonal antibody 1H2 2,2′,4,4′-tetrabromodiphenyl ether, BDE47 Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage library was incubated with Mab 1H2 (20 μg/mL). Bound phages were eluted by incubation with PBS containing 100 ng/mL BDE47. The panning procedures were then repeated twice, but the Mab concentrations of 10 and 5 μg/mL were used for the second and the third panning, respectively. 2387 VKLNDLR(1) 1 Phage display (subtractive panning) 3 PMID:24096567 Mab 1H2-BDE47 immunocomplex Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage library was firstly incubated with Mab 1H2 (20 μg/mL). Unbound phages were then incubated with Mab 1H2-BDE47 immunocomplex. The panning procedures were then repeated twice. 2388 CTELVTQLC(2) CNELVTQLC(2) CDELVRHLC(2) CQELVKHLC(1) CQELVPQLC(1) CQELVRHLC(1) 6 Phage display (subtractive panning) 3 PMID:24096567 Mab 1H2-BDE47 immunocomplex Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage library was firstly incubated with Mab 1H2 (20 μg/mL). Unbound phages were then incubated with Mab 1H2-BDE47 immunocomplex. The panning procedures were then repeated twice. 2389 WSEYDIPTPQIP(4) 1 Phage display (subtractive panning) 3 PMID:24096567 Mab 1H2-BDE47 immunocomplex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage library was firstly incubated with Mab 1H2 (20 μg/mL). Unbound phages were then incubated with Mab 1H2-BDE47 immunocomplex. The panning procedures were then repeated twice. 2390 CVHRDTIYEYC(11) CFGRDTIFEVC(4) CVGRDTVHEFC(2) R-D-T-x(2)-E 3 Phage display (subtractive panning) 3 PMID:20152791 BDE47-polyclonal antibody immunocomplex Not determined. Not determined. Not determined. CX(7-11)C phage display library Phage peptide libraries were first counter selected by incubation in the wells coated with BDE 47 PAb in the absence of BDE 47. Unbound phage peptides were then transferred to other wells that had been incubated with 10 μg/ml BDE 47. As the panning proceeded, the amount of antibody for the selection was gradually decreased to remove weak binders by increased competition. Conversely, the amount of antibody for counter selection remained constant to remove nonspecific binders. In the last panning step, BDE 47 was added to the phage library after counter selection at a concentration of 10 ng/ml to competitively remove weak binders. A total of 17 phage clones were selected for screening by phage ELISA in the presence (50 ng/ml) or absence of BDE 47. All tested phage clones reacted with the immunocomplex, showing negligible cross-reactivity with the unbound antibody. 2391 CGTFAHPQC 1 Phage display (common panning) 3 PMID:18533899 Streptavidin Biotin Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage library was incubated with streptavidin. Elution by sonication was introduced. After 25 washings with PBST and 4 with glycine-HCl (0.2 M, pH 2.2), another 100 μl of glycine-HCl were added to the microtiter plate well, and the covered plate was immersed in a sonicator water bath (50 kHz) for 10 min. Three rounds of selection were performed. A lot of phage clones were recovered. Phage-displayed peptide sequences were determined if the ELISA signal of individual clones exceeded the blank by at least fourfold. So phage clones displayed peptide CGTFAHPQC were obtained. 2392 CGTFAHPQC 1 Phage display (competitive panning) 3 PMID:18533899 Streptavidin Biotin Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage library was incubated with streptavidin. After 25 washings with PBST and 4 with glycine-HCl (0.2 M, pH 2.2), 100 μl of 0.1 mM biotin was used for eluting. A lot of phage clones were recovered. Phage-displayed peptide sequences were determined if the ELISA signal of individual clones exceeded the blank by at least fourfold. So phage clones displayed peptide CGTFAHPQC were obtained. 2393 VVFTSWQDYLFWV 1 Phage display (common panning) 4 PMID:24225950 Proprotein convertase subtilisin/kexin type 9, PCSK9 Low-density lipoprotein receptor 2W2M,2W2N,2W2O,2W2P,2W2Q,3BPS,3GCW,3GCX,3M0C,3P5B,3P5C, Not determined. Linear-lib and Cyclic-lib M13 phage display library pool Phage pools of Linear-lib and Cyclic-lib library were cycled through rounds of binding selections. For round one, 130 nM of biotinylated PCSK9 was incubated with the phage library pool, and the phage-PCSK9 complex was captured by 200μl of Dynabeads® MyOne Streptavidin. For round two, the protocol was the same as round one except for using 65 nM of biotinylated PCSK9 and 100μl of Dynabeads. For round three, 26 nM of biotinylated PCSK9 was incubated with the amplified phage from the previous round, and the phage-PCSK9 complex was captured by NeutrAvidin-coated plates. Round four was identical with round three except for using Streptavidin-coated plates to capture biotin–PCSK9–phage complex. After four rounds of binding selection, several clones gave modest binding signals. Individual phage clones were analyzed in a high throughput spot phage ELISA using plate-immobilized PCSK9 as the target. Nonspecific binding was defined as phage particle binding to NeutrAvidin. Clones with phage binding signal to PCSK9 over 0.5 and signal/noise ratio of 5 were considered to be positive clones and were subjected to DNA sequence analysis. The peptides derived from these clones were synthesized and tested in the HepG2 LDL receptor degradation assay. Pep2 (VVFTSWQDYLFWV)derived from clone 2 was able to inhibit PCSK9-mediated LDL receptor degradation with low potency. 2394 QSHYRHISPAQV(20) TLAYARAYMVAP(2) QSHYRHISPDQV(1) QSHYRHISPARV(1) KSHYRHISPAKV(1) HHGHSPNVSQVR(1) GSFSTQVGSLHR(1) HTGTQSYVPRLR(1) ATPQNDLKTFPH(1) TQPETDLLRVQF(1) CITWPPTGLTYP(1) TFLETGPIYADG(1) LVPPTHRHWPVT(1) APPGNWRNYLMP(1) DNYSNYVPGTKP(1) SVSVGMKPSPRP(1) SLPNPFSVSSHG(1) YVHNPYHLPNPP(1) CRRLHTYLGPVT(1) 19 Phage display (competitive panning) 4 PMID:12898626 Soluble 37/48kDa oligomer formation of Aβ(1-42) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A commercially available phage-displayed peptide library (PhD12, New England Biolabs) was screened for peptide variants with affinity for synthetic bio-D-Aβ (1-42) immobilized on the surface of streptavidin-coated tubes. The phage-peptide suspension was transferred into a streptavidin-coated tube. To displace any streptavidin-binding phages, biotin (0.1 mM) was added. Eventually phages were eluted by 10-min incubation with glycine-HCl (0.8 mL, 0.2M, pH 2.2) and neutralized with Tris-HCl (0.2 mL, 1M, pH 9.1). After four rounds of selection and amplification, 39 randomly picked phages were subjected to DNA sequence analysis of the 5'-end of the gene coding for the minor coat protein. Upon translation into amino acid sequences, 23 of the resulting peptides showed strong sequence similarity. 2395 SEVGCRAGPLQLCEKYF 1 Phage display (common panning) 4 PMID:11075354 Insulin-like growth factor-binding protein 1, IGFBP-1 Insulin-like growth factor I, IGF-I 2DSQ, Not determined. SGTACX2GPX4CSLAGSP phage display library For the first three rounds of selection, IGFbp-1 was covalently linked to biotin using EZ-Link NHS-SS-biotin (Pierce). The biotin-SS-IGFbp-1 was first allowed to bind to immunosorbent plates coated with NeutrAvidin (Pierce). The fourth round of binding selection was carried out on immunosorbent plates directly coated with IGFbp-1. 2396 KDPVCGEGPLMRICERLF 1 Phage display (common panning) 4 PMID:11075354 Insulin-like growth factor-binding protein 1, IGFBP-1 Insulin-like growth factor I, IGF-I 2DSQ, Not determined. X4CX2GPX4CX4 phage display library For the first three rounds of selection, IGFbp-1 was covalently linked to biotin using EZ-Link NHS-SS-biotin (Pierce). The biotin-SS-IGFbp-1 was first allowed to bind to immunosorbent plates coated with NeutrAvidin (Pierce). The fourth round of binding selection was carried out on immunosorbent plates directly coated with IGFbp-1. 2397 EVDGRWWIVETFLAKWDHMA 1 Phage display (common panning) 4 PMID:11075354 Insulin-like growth factor-binding protein 1, IGFBP-1 Insulin-like growth factor I, IGF-I 2DSQ, Not determined. X20 M13 phage display library For the first three rounds of selection, IGFbp-1 was covalently linked to biotin using EZ-Link NHS-SS-biotin (Pierce). The biotin-SS-IGFbp-1 was first allowed to bind to immunosorbent plates coated with NeutrAvidin (Pierce). The fourth round of binding selection was carried out on immunosorbent plates directly coated with IGFbp-1. 2398 QGAMECEVVPRGVMCVLDSK 1 Phage display (common panning) 4 PMID:11075354 Insulin-like growth factor-binding protein 1, IGFBP-1 Insulin-like growth factor I, IGF-I 2DSQ, Not determined. X5CX8CX5 phage display library For the first three rounds of selection, IGFbp-1 was covalently linked to biotin using EZ-Link NHS-SS-biotin (Pierce). The biotin-SS-IGFbp-1 was first allowed to bind to immunosorbent plates coated with NeutrAvidin (Pierce). The fourth round of binding selection was carried out on immunosorbent plates directly coated with IGFbp-1. 2399 EVCVQIEWWCSH 1 Phage display (common panning) 4 PMID:11075354 Insulin-like growth factor-binding protein 1, IGFBP-1 Insulin-like growth factor I, IGF-I 2DSQ, Not determined. X2CX6CX2 phage display library For the first three rounds of selection, IGFbp-1 was covalently linked to biotin using EZ-Link NHS-SS-biotin (Pierce). The biotin-SS-IGFbp-1 was first allowed to bind to immunosorbent plates coated with NeutrAvidin (Pierce). The fourth round of binding selection was carried out on immunosorbent plates directly coated with IGFbp-1. 2400 YMDSGCAQGWEEYCMWECS 1 Phage display (common panning) 3 PMID:11075354 Bovine insulin-Like growth factor-binding protein 2, bIGFbp-2 Insulin-like growth factor I, IGF-I Not determined. Not determined. X5CX8CX5 phage display library BlGFbp-2 was coated directly onto immunosorbant plates. The library was subjected to three rounds of selection, using 100 mM HC1 elution, followed by neutralization. Phage from round 1 were amplified in E. coli before use in round 2. The eluted phage from round 2 were not amplified before use in round 3. 2401 VTQESCCWWEDMVGMECGRLE 1 Phage display (common panning) 3 PMID:11075354 Bovine insulin-Like growth factor-binding protein 2, bIGFbp-2 Insulin-like growth factor I, IGF-I Not determined. Not determined. X5CX9CX4 phage display library BlGFbp-2 was coated directly onto immunosorbant plates. The library was subjected to three rounds of selection, using 100 mM HC1 elution, followed by neutralization. Phage from round 1 were amplified in E. coli before use in round 2. The eluted phage from round 2 were not amplified before use in round 3. 2402 RDCDTSCGH 1 Phage display (common panning) 3 PMID:11075354 Bovine insulin-Like growth factor-binding protein 2, bIGFbp-2 Insulin-like growth factor I, IGF-I Not determined. Not determined. X2CX3CX2 phage display library BlGFbp-2 was coated directly onto immunosorbant plates. The library was subjected to three rounds of selection, using 100 mM HC1 elution, followed by neutralization. Phage from round 1 were amplified in E. coli before use in round 2. The eluted phage from round 2 were not amplified before use in round 3. 2403 SNCFWDGATVVCAQ TDCTGPWVWVWCPQV 2 Phage display (common panning) 3 PMID:11075354 Bovine insulin-Like growth factor-binding protein 2, bIGFbp-2 Insulin-like growth factor I, IGF-I Not determined. Not determined. X2CX8CX2 phage display library BlGFbp-2 was coated directly onto immunosorbant plates. The library was subjected to three rounds of selection, using 100 mM HC1 elution, followed by neutralization. Phage from round 1 were amplified in E. coli before use in round 2. The eluted phage from round 2 were not amplified before use in round 3. 2404 DGCAWDGVQMVDCTG 1 Phage display (common panning) 3 PMID:11075354 Bovine insulin-Like growth factor-binding protein 2, bIGFbp-2 Insulin-like growth factor I, IGF-I Not determined. Not determined. X2CX10CX2 phage display library BlGFbp-2 was coated directly onto immunosorbant plates. The library was subjected to three rounds of selection, using 100 mM HC1 elution, followed by neutralization. Phage from round 1 were amplified in E. coli before use in round 2. The eluted phage from round 2 were not amplified before use in round 3. 2405 DALRRTRGGAAAGFAVHGSL GNARLHVVARDHTGFAVLGT LTTDLMHCSGFALSPGCPQI MNGKVGWAIHAREAVGTREG DDSSHTGWAVLDLGLRQPVF GDTHIGPDSGQREHVGFAIL DALRRTRGGAAAGFAVHGSL GNARLHVVARDHTGFAVLGT LTTDLMHCSGFALSPGCPQI MNGKVGWAIHAREAVGTREG DDSSHTGWAVLDLGLRQPVF GDTHIGPDSGQREHVGFAIL G-[FW]-A 12 Phage display (common panning) PMID:19653209 Anti-gp120 monoclonal antibody GV4H3 Surface protein gp120, SU Not determined. Not determined. X20 FUSE5-based phage display library No particular motif can be identified for the first six sequences. However, the common GFA (GWA) tripeptide can be recognized for the last six sequences. In this case, homologies with the sequence 221-226 (AGFAIL) of HIV-1 gp120 can be identified. 2406 TLPSPLALLTVH(2) SPPSNLIPPTLR(2) ATWSHHLSSAGL(1) QTRPHHLRSATQ(1) 4 Phage display (common panning) 3 PMID:18195017 Death-associated protein kinase 1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) For each round of panning, the bound phage particles were eluted by ATP incubation and neutralized with 15 μl of 1 M Tris-HCl (pH 9.1). 2407 LPFEEHLRRPVG(1)[0.452 ± 0.071] SNLPQSWPPHQW(1)[NT] QGAWHPSRTHFL(1)[NT] TQPPIPPSRTLH(1)[NT] TFGLWQPGVQST(1)[NT] WHWSWFSHFPSA(1)[NT] 6 Phage display (common panning) 3 PMID:18195017 Death-associated protein kinase 1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The ability of purified DAPK-1 to bind isolated phage 12-mers was further assessed using synthesized biotinylated peptide. Synthetic peptide Φ (LPFEEHLRRPVG), the naturally occurring peptide sequence in MAP1B itself (1B, IVTEEHLRRAIG), or control peptides (NR) were added to the wells with DAPK-1. Peptide binding activity (RLU, relative light units) were reproduced from the graph and shown. The values of the peptide 1B and NR were 0.939 ± 0.168 and 0.018, respectively. NT denotes not tested. For each round of panning, the bound phage particles were eluted by acid incubation and neutralized with 15 μl of 1 M Tris-HCl (pH 9.1). 2408 CPNNTISLC[1.246 ± 0.024] CPLTTKTLC[1.448 ± 0.054] 2 Phage display (competitive panning) 3 PMID:24447276 Globotriaosylceramide, Gb3 Shiga toxin, Shiga toxin subunit A and Shiga toxin subunit B Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Phage ELISA Gb3-binding assay was performed. The absorbance of the end product was measured at a wavelength of 492 nm using an ELISA microplate reader. Raw absorbance data were corrected by subtraction of the values obtained from an OPD-containing blank. This assay was performed in triplicate. The absorbance values were determined. Data were reproduced from the graph and shown as means ± the standard deviations. The panning procedure was repeated for a total of three rounds for the Ph.D.-C7C library. For each round of panning, bound phages were eluted with Gb3. The highest competition percentages for binding to Gb3 between the toxins and peptides (CPNNTISLC and CPLTTKTLC) were observed at peptide concentrations ranging from 25 to 100 μg. The Stx cytotoxic activity neutralization assay indicated that when Escherichia coli O157 : H7 cell-free filtrate was applied to Vero cells, the inhibition of cell death was similar for the two peptides at the different concentrations assayed. Besides, the inhibition of lethality assay by using peptides CPNNTISLC and CPLTTKTLC was performed. The peptide CPNNTISLC partially inhibited the lethality caused by Stx1 in mice, whereas peptide CPLTTKTLC abolished the lethality induced by Stx1. In contrast, none of the peptides assayed inhibited the lethality caused by the Stx2 toxin in mice. Moreover, none of the peptides themselves induced lethality in mice. 2409 DGYYRSNNAGTP[0.504 ± 0.039] DNYTRYDYMDIP[0.137 ± 0.005] SAPRHNVPDNPR[2.093 ± 0.029] SSQPHTVFDPSK[0.429 ± 0.021] 4 Phage display (competitive panning) 3 PMID:24447276 Globotriaosylceramide, Gb3 Shiga toxin, Shiga toxin subunit A and Shiga toxin subunit B Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Phage ELISA Gb3-binding assay was performed. The absorbance of the end product was measured at a wavelength of 492 nm using an ELISA microplate reader. Raw absorbance data were corrected by subtraction of the values obtained from an OPD-containing blank. This assay was performed in triplicate. The absorbance values were determined. Data were reproduced from the graph and shown as means ± the standard deviations. The panning procedure was repeated for a total of three rounds for the Ph.D.-12 library. For each round of panning, bound phages were eluted with Gb3. The highest competition percentages for binding to Gb3 between the toxins and the peptide SAPRHNVPDNPR were observed at peptide concentrations ranging from 25 to 100 μg. The Stx cytotoxic activity neutralization assay indicated that when Escherichia coli O157 : H7 cell-free filtrate was applied to Vero cells, the inhibition of cell death was similar for the three peptides at the different concentrations assayed. The only exception was the inhibition caused by the peptide SAPRHNVPDNPR, which was higher than the inhibition caused by peptides CPNNTISLC and CPLTTKTLC at the concentration of 50 μg per well. Besides, the inhibition of lethality assay by using the peptide SAPRHNVPDNPR was performed. The peptide partially inhibited the lethality caused by Stx1 in mice. In contrast, it didn't inhibit the lethality caused by the Stx2 toxin in mice. Moreover, it didn't induce lethality in mice. 2410 TEPSTRGSWKFW(13) SVPQESTIRATK(5) SGHTHYYPAETN(4) LCNTYLPLARST(3) VPILAHPSTGHS(2) SENRKVPFYSHS 5 Phage display (common panning) 5 PMID:24468276 The Domain III of the Japan Encephalitis Virus Envelope Protein, E DIII Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The homologous alignment of peptides displayed by frequently encountered phages was analyzed, resulting in the identification of a highly conserved 12 amino acid peptide sequence that was also synthesized and named P3 (SENRKVPFYSHS). The inhibitory ability of the peptides against JEV infection were determined by plaque assays, qRT-PCR and WB. The plaque assay results showed that P3 significantly inhibited JEV infection in a concentration-dependent manner, whereas P1 (TEPSTRGSWKFW) inhibited JEV infection only at high concentrations. P2 (SVPQESTIRATK) did not show any inhibitory effect. Besides, P3 bound to E DIII with a Kd of 6.06 × 10 (-6) M, indicating that P3 has a higher affinity for E DIII. Also P3 interfered with JEV binding to the cell receptor. Eventually A model of the E DIII/P3 complex was developed by molecular modeling based on the available E DIII structure. 2411 IPTLPSS(3) SSPSTSR(3) TRAPWPP(2) YRAPWPP(2) LPSL*NI(1) SSPATSR(1) FSESTQAS(1) STYTSVS(1) APQPPEP(1) TPSPHRV(1) WAPTPSR(1) STPSWWA(1) AITRSPA(1) FSPPRFY(1) YMTPLRP(1) TPWHPLA(1) 16 Phage display (subtractive panning) 4 PMID:24480037 BCR/ABL-expressing NIH3T3 mouse fibroblast cells Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage-display linear heptapeptide library Ph.D 7.0 was used to pan NIH3T3 clones, using a combination of negative/positive panning in each round. All planning stages were performed at RT in PBS/3% BSA. Phages were preincubated in 10 volumes PBS/3% prior to each round of panning and then used to first pan the control cells (3T3C.pCVT). The supernatant was removed and used to negatively pan a second set of 3T3C.pCVT cells. This phage-depleted supernatant was used to positively pan against the BCR/ABL+ cells (3T3B.pGD210). This panning strategy was performed in two independent experiments, with 10 phages initially sequenced following three rounds of panning in each experiment. Although no dominant motif was observed, the combined analysis showed a predominance of ligands containing proline, serine and threonine residues, with 71% containing at least one proline residue and 57% containing serine or threonine residues. Round 4 panning was performed and 20/27 phages were sequenced respectively. No significant difference was observed between the two experiments in terms of sequence dominance and thus the 47 sequences were analysed collectively to look for predominant motifs. Again, a similar enrichment of phage peptides containing serine, threonine and proline residues was observed. 2412 TNLTLAS(8) GALPNNL(6) SLAVSRS(4) SEIVDNH(1) NVNSTSF(1) SPDTVQK(1) GNRLSAD(1) LGFREKE(1) TQVYHPM(1) ANHQSAN(1) TNSSFHK(1) NTVIYQK(1) HVQYWQF(1) VSVNSRT(1) RLLNPWI(1) 15 Phage display (common panning) 5 PMID:24512376 Amorphous Ni3B Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2413 LEQTPMF(1) ELTQISS(1) SDPQTHT(1) TPPLLSP(1) MNHAESY(1) VPSLTPT(1) VPIPYLP(1) DPYNRIN(1) RTFDAIS(1) YELVLPK(1) ETFPARG(1) GPVNHQL(1) LNHVLPA(1) HAMRTEP(1) ATSTAHA(1) ANHQSAN(1) SYTKLHL(1) SPPKSNA(1) SASKVHN(1) SPSTHWK(1) WNAKYTL(1) YQVVPAR(1) GDPKAAR(1) GDHSRHK(1) AGLPKHQ(1) STFNSRV(1) VHTNPSR(1) GASATRT(1) 28 Phage display (common panning) 5 PMID:24512376 Crystalline Ni3B Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2414 AGNWTPI(7)[0.623 ± 0.034] RPTSHQL(1)[0.486 ± 0.021] QWTPSHP(1)[0.589 ± 0.017] APDTKTQ(1)[0.561 ± 0.048] 4 Phage display (common panning) 4 PMID:24530908 Fibroblast growth factor 1, FGF-1 FGFR-1, FGFR-2, FGFR-3 and FGFR-4 1DJS,1E0O,3CU1,3OJ2,3OJM,4J23,1EVT,3OJV,1RY7, Not determined. Ph.D.-7 phage display library (X7) ELISA Binding abilities of phage clones to aFGF were determined by ELISA. The absorbance at 370 nm was measured. Data were reproduced from the graph and expressed as means ± S.D. The AP8 peptide (AGNWTPI), with amino acids identical to FGFR1, may bind aFGF via electrostatic interactions and may interrupt aFGF binding to FGFR1. Results of competitive inhibition assay indicated that the synthetic peptide AP8 and the corresponding phage clone A8 thus compete for the same binding site, and the binding of phage A8 to aFGF was mediated by the peptide AP8. Besides, the proliferation of MDA-MB-231, MCF-7 and HUVEC cells stimulated with aFGF could be inhibited by AP8 in a dose-dependent manner, while AP8 had little inhibitory effect on Cos-7 cells that do not express aFGF receptors. AP8 arrests aFGF-induced cells at the G0/G1 phase via Cyclin D1. aFGF significantly stimulated the phosphorylation of Erk1/2 and Akt signal molecules, while the pretreatment of cells with AP8 for 30 min prior to aFGF stimulation resulted in significant blockage of the activation of the both signal molecules in a dose-dependent manner. AP8 counteracts the regulatory effect of aFGF on PA2G4 and PCNA expression. 2415 ERNSVSPS DRNPPLPS EPFQVGDQ VHNTTSSS [DE]-R-N-x(3)-P-S 4 Phage display (common panning) 3 PMID:24533565 Prostate-specific antigen Not determined. Not determined. Not determined. f8-8mer phage display library (X8) 2416 KPPKRKADRWVP LGADRTRDRRHN TERRGSSKYPKR SHLPKVRSPTQK NKWPLAHSQKKR LKPNKPRMHLDI GGSGTSRTPILG SRSSGLKKQYHK 8 Phage display (common panning) 4 PMID:24535972 Calcium silicate hydrate 0.66, C-S-H 0.66 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The pH during the panning experiment was 8.9. 2417 YHPNGMNPYTKA LPGRAHDPWKVP SATNGSLTRPVH GTTTLNHNYSAK DGNTAARMATLK VPRSMAATHSTF ANHLSGNNYGIS DSASTQFTRADS 8 Phage display (common panning) 4 PMID:24535972 Calcium silicate hydrate 1.0, C-S-H 1.0 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The pH during the panning experiment was 11.6. 2418 LPNHLGAILRAD SSLHSSHHEMNK GTTTLNHNYSAK KFNPGNSEWQRT SKHDTPTYINTV IDRVTSRDPAMN VDTVKHELATYR SATNGSLTRPVH YHPNGMNPYTKA VPRSMAATHSTF GGSIAASELEYY IFAALDYNLGRH 12 Phage display (common panning) 4 PMID:24535972 Calcium silicate hydrate 1.5, C-S-H 1.5 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The pH during the panning experiment was 12.5. 2419 LPNHLGAILRAD YHPNGMNPYTKA GGSIAASELEYY SATNGSLTRPVH TLRVPPNPNMNV VPRSMAATHSTF IEAQYNHSNRFP MTFDTVSNIYKM HAAGIRDNQRLG DGNTAARMATLK TCAKATSTPPLS 11 Phage display (common panning) 4 PMID:24535972 Calcium silicate hydrate 1.7, C-S-H 1.7 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The pH during the panning experiment was 12.5. 2420 TLHDLTRGQRTT(2)[12.748 ± 0.187] ANPYSSTAKPAG(2)[6.660 ± 0.062] RPLTISSAADHF(1)[14.448 ± 0.152] EAHVMHKVAPRP(1)[6.847 ± 0.124] SEPPKAHGVLSS(1)[4.256] LPSPPRIPGHKL(1)[9.473 ± 0.187] NSAAVRAYSPPS(1)[13.522 ± 0.428] QHANHQAWNNLR(1)[8.208 ± 0.028] SHNNSPLSYKPS(1)[9.998 ± 0.097] NFMESLPRLGMH(1)[12.347 ± 0.553] 10 Phage display (competitive panning) 3 PMID:24607635 Salivary pellicle Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay All the 10 novel salivary pellicle-binding peptides were analyzed for binding to saliva-coated HA. The amount of bound peptide (μg) to saliva-coated HA was reproduced from the graph and shown as means ± S.D (N = 3). In total three rounds of panning were performed. In the first round of panning the pellet, washing was done with 1 ml saliva buffer containing 0.1% Tween-20. In the second and third round of panning the pellet, containing phages bound to saliva-coated HA, was washed ten times with saliva buffer containing 0.5% Tween-20. Binding of SPBP10 (NSAAVRAYSPPS) to HA was evaluated in saliva buffer, containing 1 mM CaCl2. To determine whether SPBP 10 exerts antifouling activity, the effect of treatment of HA with SPBP 10 on the adherence of S. gordonii was evaluated in an in vitro adherence model. In the absence of SPBP10, approximately 7.7e5 CFU/ml S gordonii adhered to the surface of the HA discs. After treatment of HA discs with SPBP 10, the number of adhered bacteria decreased significantly to approximately 3.8e5 CFU/ml ( p ≤ 0.05). Considerably fewer bacteria adhered to saliva-coated discs than to HA discs. After treatment with SPBP 10 the number of adhered bacteria was even lower, but no statistical significance was reached. 2421 AARPVAL ASRPPAP SAGHPKY 3 Phage display (competitive panning) PMID:20412822 Lipopolysaccharide, LPS Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The wells of a microtiter plate were coated with LPS or lipid A by treating with 50 μl of 100 μg/ml LPS or lipid A in phosphate-buffered saline. After washing with 0.05% Tween-20 in PBS (PBST), the wells were blocked with 200 μl of 1% skim milk at 25 °C for 2 h and washed with PBST. Then, 0.1 ml of M13 phagee expressing 7- or 12-residue peptides encoded by random sequences of nucleotides fused with DNA for phage coat protein was added to the wells. After washing off free phage particles, the wells were treated with 0.1 ml of PBS containing 100μg/ml of LPS or lipid A to elute the phages bound to the ligand. Using a 7 residues library, 3 ELISA positive clones from 18 clones were obtained. Among the 7-residue peptides, Li1 (AARPVAL) and Li2 (ASRPPAP) have a common motif, AXRPXA, but this sequence was not found in Li3 (SAGHPKY). Among these 3 peptides, we synthesized AARPVALGGSC by adding 3-residue spacer peptide (GGS) and cysteine at the C-terminus of Li1 . Its binding affinity to LPS, as measured by Biacore, was very weak. 2422 MGHRPPSPSLSG(4) 1 Phage display (competitive panning) PMID:20412822 Lipopolysaccharide, LPS Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The wells of a microtiter plate were coated with LPS or lipid A by treating with 50 μl of 100 μg/ml LPS or lipid A in phosphate-buffered saline. After washing with 0.05% Tween-20 in PBS (PBST), the wells were blocked with 200 μl of 1% skim milk at 25 °C for 2 h and washed with PBST. Then, 0.1 ml of M13 phagee expressing 7- or 12-residue peptides encoded by random sequences of nucleotides fused with DNA for phage coat protein was added to the wells. After washing off free phage particles, the wells were treated with 0.1 ml of PBS containing 100μg/ml of LPS or lipid A to elute the phages bound to the ligand. Four ELISA positive clones were obtained from 18 clones and these 4 clones had the same sequence. 2423 KNYSSSISSIRA 1 Phage display (competitive panning) PMID:20412822 Lipopolysaccharide, LPS Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The wells of a microtiter plate were coated with LPS or lipid A by treating with 50 μl of 100 μg/ml LPS or lipid A in phosphate-buffered saline. After washing with 0.05% Tween-20 in PBS (PBST), the wells were blocked with 200 μl of 1% skim milk at 25 °C for 2 h and washed with PBST. Then, 0.1 ml of M13 phagee expressing 7- or 12-residue peptides encoded by random sequences of nucleotides fused with DNA for phage coat protein was added to the wells. After washing off free phage particles, the wells were treated with 0.1 ml of PBS containing 100μg/ml of LPS or lipid A to elute the phages bound to the ligand. In a 12-residue library, one positive clone (displaying KNYSSSISSIRA) was obtained from 12 clones. 2424 WPHFHHLRVPPV[370] DQRVLPSTFAAD[ND] DRLHHHRHSWKY[ND] HIHKHTVFLNSP[ND] HLKWLPHHRQPM[ND] HYFTWWPHRNPH[ND] KPISHHPHHRAW[ND] QYKTQHIYGYGP[ND] TFTHHRHYPKVV[ND] TPHLHMWHAHKR[ND] WHRHTLAPHSHP[ND] WHRIQIPPAPIL[ND] WPHNWWPHFKVK[ND] WPYHPHRHPEPL[ND] 14 Phage display (common panning) 3 PMID:12659965 Calcium silicate hydrate 1.7, C-S-H 1.7 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR) The apparent variation in the association and dissociation rates of individual phage-displayed peptides on the LA chip suggests the potential of strategically using SPR technology to rapidly rank the LPS/LA binding affinities of phage-displayed peptides with high throughput. The calculated apparent KD (nM) is 370 nM for LA/11 (WPHFHHLRVPPV). ND denotes not determined. 2425 KSLSRHDHIHHH[0.0055] HFKHKHPEPPGR[ND] YPFHHKHWQRPD[ND] LGRHTHHFWHYP[ND] YPWHSRHAPRVL[ND] YPWTHHHSRWDL[ND] FTRHHHPGFWWN[ND] WGFHYPRYHTLK[ND] WWTPWRLHGGPH[ND] WHKSPKLPLSPV[ND] HLKMFHWSVPPN[ND] APMHKYHSWHKR[ND] WPHQKLHLMRHS[ND] HTYSVYPPRDFK[ND] HILNQRPIYLGT[ND] 15 Phage display (common panning) 3 PMID:12659965 Lipopolysaccharide, LPS Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR) The apparent variation in the association and dissociation rates of individual phage-displayed peptides on the LA chip suggests the potential of strategically using SPR technology to rapidly rank the LPS/LA binding affinities of phage-displayed peptides with high throughput. The calculated apparent KD (nM) is 0.0055 nM for LPS/10 (KSLSRHDHIHHH). ND denotes not determined. 2426 QTHQELQTLTTS[0.98] HLNQRNQPPDGA[0.139] VPTHNKGLPEPV[0.301] 3 Phage display (common panning) 4 PMID:24759178 IGVH1 template protein Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR) The SPR assay of the peptides binding to the human Fv templates was performed. The binding affinity (KD, μM) between ligand peptides and human FV templates was shown. The value shows the representative data of two-time experiments. 2427 WSEYDIPTPQIP[6.29] HIPWLHAMSVMK[0.0605] IAFNTPPHHPLR[0.00803] GAMHLPWHMGTL[ND] NRPDSAQFWLHH[ND] HFAYWWNGVRGP[ND] SWMPHPRWSPQH[ND] HGWTLWHMALTS[ND] HLWLPNGAFRSW[ND] 9 Phage display (common panning) 4 PMID:24759178 IGVH3 template protein Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR) The SPR assay of the peptides binding to the human Fv templates was performed. The binding affinity (KD, μM) between ligand peptides and human FV templates was shown. The value shows the representative data of two-time experiments. ND denotes not determined. 2428 HFAYWWNGVRGP 1 Phage display (common panning) 4 PMID:24759178 IGVH4 template protein Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) 2429 HFPWLDRSAVDM[0.0587] QTHQELQTLTTS[ND] 2 Phage display (common panning) 4 PMID:24759178 IGVH6 template protein Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR) The SPR assay of the peptides binding to the human Fv templates was performed. The binding affinity (KD, μM) between ligand peptides and human FV templates was shown. The value shows the representative data of two-time experiments. ND denotes not determined. 2430 HFWFVPALQGAP[0.047] TPTKSSFYLPPN[0.0141] HFAYWWNGVRGP[ND] HIRWDVNHNSMS[ND] SHPWYIGPHPNA[ND] HSQSLLRLWQVY[ND] 6 Phage display (common panning) 4 PMID:24759178 IGVK1 template protein Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR) The SPR assay of the peptides binding to the human Fv templates was performed. The binding affinity (KD, μM) between ligand peptides and human FV templates was shown. The value shows the representative data of two-time experiments. ND denotes not determined. 2431 HFAYWWNGVRGP WHWSWNPRFAPS 2 Phage display (common panning) 4 PMID:24759178 IGVK2 template protein Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) 2432 NIYRDTHLEKFA VHYEDPTQLFTS 2 Phage display (common panning) 4 PMID:24759178 IGVK3 template protein Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) 2433 WHWNWMRLDPMT[2.48] 1 Phage display (common panning) 4 PMID:24759178 IGVK4 template protein Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR) The SPR assay of the peptides binding to the human Fv templates was performed. The binding affinity (KD, μM) between ligand peptides and human FV templates was shown. The value shows the representative data of two-time experiments. 2434 WHWNWMRLDPMT 1 Phage display (common panning) 4 PMID:24759178 IGVK5 template protein Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) 2435 HSWGWVAMNTQT[0.646] HFAYWWNGVRGP[ND] 2 Phage display (common panning) 4 PMID:24759178 IGVK6 template protein Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR) The SPR assay of the peptides binding to the human Fv templates was performed. The binding affinity (KD, μM) between ligand peptides and human FV templates was shown. The value shows the representative data of two-time experiments. ND denotes not determined. 2436 SLMVPSHSEPIR[0.782 ± 0.010] HYNAWNGWRFWT[0.628 ± 0.031] FHWTWQFPYTST[0.683 ± 0.031] SEFHRSWDMETN[0.366 ± 0.124] QAMYSKYPMINV[0.202 ± 0.011] FHRSHWYQTWVP[0.989 ± 0.010] HLWSNWQFWNMA[1.222 ± 0.015] 7 Phage display (common panning) 4 PMID:24759178 IGVK3 template protein Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Phage ELISA was performed for measuring peptide binding to the IGVK template. Absorbance at 405 nm was determined, reproduced from the graph and expressed as means ± S.D. The data shows the representative data of two-time experiments. 2437 WLAYPGAVSYR(19/20)[0.916][678 ± 23] 1 Phage display (common panning) 3 PMID:12351647 Ephrin type-A receptor 2 Type A ephrins Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA,Surface plasmon resonance (SPR) Phage ELISA was performed to measure the affinity of phage to EphA2. Absorbance at 405 nm was determined, reproduced from the graph and shown. Besides, equilibrium binding data obtained by surface plasmon resonance indicate that the SWLAYPGAVSYR peptide binds to EphA2 with certain affinity (KD = 678 nM ± 23). Nineteen of 20 individual phage clones from the pool eluted with low pH bind specifically to EphA2 Fc. All 19 clones display the same peptide: SWLAYPGAVSYR (SWL peptide). 2438 SWLAYPGAVSYR(7/10)[0.916][678 ± 23] YSAYPDSVPMMS(2/10)[0.856][186 ± 7] 2 Phage display (competitive panning) 3 PMID:12351647 Ephrin type-A receptor 2 Type A ephrins Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA,Surface plasmon resonance (SPR) Phage ELISA was performed to measure the affinity of phage to EphA2. Absorbance at 405 nm was determined, reproduced from the graph and shown. Besides, equilibrium binding data obtained by surface plasmon resonance indicate that the YSAYPDSVPMMS peptide binds to EphA2 with higher affinity (KD = 186 nM ± 7) than the SWLAYPGAVSYR peptide (KD = 678 nM ± 23). For each round of biopanning, phages remaining bound after washing were eluted with ephrin-A1 Fc. Nine of 10 phage clones from the pool eluted with ephrin-A1 bind specifically to EphA2 (data not shown). Seven of these clones display the SWLAYPGAVSYR peptide (SWL peptide) and two display the peptide YSAYPDSVPMMS (YSA peptide). The YSA peptide targets EphA2 on the cell surface and stimulates activation of the receptor. Thus, the YSA peptide is an agonist for EphA2. Besides, the YSA and SWL peptides inhibit ephrin-A binding to EphA2. To further characterize the binding site of the YSA and SWL peptides, competition experiments with phage displayed peptides were performed. Synthetic SWL peptide competes with YSA phage bound to immobilized EphA2, and conversely YSA peptide competes with SWL phage. This indicates that the YSA and SWL peptides bind to the same or overlapping sites on EphA2. 2439 HAIYPRH[1.10 ± 0.08] AKPGYLS[>1.9] ALTPTPP[>1.9] ANYPREP[>1.9] ATTVPAS[>1.9] LTTHYKL[>1.9] NLVNLLP[>1.9] KCCFPAQ[NT] 8 Phage display (in vivo) 3 PMID:22463021 Colonic adenomas Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Flow Cytometry Binding to isolated colonic epithelial cells from adenomas for each phage clone was analyzed using flow cytometry. A common 7-mer phage that expresses the sequence HAIYPRH is a known contaminant and was run to ensure the validity of the assay. The T/B ratio for clone HAIYPRH was found to be 1.10 ± 0.08, indicating minimal binding from a nonspecific clone, as expected. Six phage clones with a T/B ratio (target to wild-type phage) >1.9 were identified as ALTPTPP, NLVNLLP, ANYPREP, ATTVPAS, LTTHYKL, and AKPGYLS. ND denotes not determined. A seven-month old CPC, Apc mouse, was injected via tail vein with the phage display library. A total of three rounds of phage biopanning were performed, with amplification of the recovered eluate after each biopanning round. After three rounds of in vivo phage biopanning, 42 individual phage clones were sequenced from the third round of biopanning and were individually amplified and conjugated to 5'-FITC. 2440 SLDSTHTHAPWP(3)[1.687 ± 0.075] TLADTHTHRPWT(2)[1.523 ± 0.117] LALDTPSHRPWT(2)[1.486 ± 0.078] FWGVTPHGELRS(1)[1.491 ± 0.103] 4 Phage display (subtractive panning) 4 PMID:24478253 Hepatocellular carcinoma cell line HepG2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Phage ELISA was performed to evaluate the affinity of eight phage clones to HepG2 cells. Plates were read at 450 nm. Phage clones displaying IRPs and PBS treatments were used as negative controls. Triplicate determinations were performed. Absorbance at 450 nm was reproduced from the graph and expressed as means ± S.D. The absorbances of phage clones displaying IRPs and PBS treatments were 0.031 and 0.025 ± 0.006, respectively. HepG2 cells and HEK293 cells were separately seeded into six-well. The phage library was incubated with HEK293 cells. After incubation, culture medium containing the unbound phages was transferred to HepG2 cell monolayer cultures and incubated. Four rounds of biopanning were performed. In phage ELISA, the PC28 clone (displaying SLDSTHTHAPWP) appeared to bind most effectively to HepG2 cells than the other clones. Immunofluorescence assay further evaluated the affinity of the PC28 phage clone binding to HepG2 cells. Besides, the synthetic peptide HCSP4, SLDSTHTHAPWP, can compete with the binding of the PC28 phage clone to HepG2 cells in competitive inhibition assay, which indicated that the binding of phage PC28 to target cells was mediated by the peptide HCSP4. The binding specificity of the FITC-conjugated HCSP4 to HepG2 cells was further validated by flow cytometry analysis. 2441 HPWIPKR[0.330 ± 0.017] WPWQHHR[0.493 ± 0.031] WPWHHVR[0.417 ± 0.029] WPWHNHR[0.514 ± 0.050] VLRSDFK[0.048 ± 0.035] W-P-W-x(3)-R 5 Phage display (common panning) 3 PMID:24573486 Envelope glycoprotein E2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA In phage ELISA, the absorbance values were measured at 490 nm (A490). The values shown were reproduced from the graph and expressed as means ± S.D. Twenty-five plaques were randomly selected from the third round phages, and their binding reactivity to the HCV E2661 protein was determined individually using an ELISA. Clone no. 3 (displaying VLRSDFK) had the lowest affinity to E2661 protein. Four clones with the highest affinities (nos. 7, 11, 17 and 18 displaying HPWIPKR, WPWQHHR, WPWHHVR and WPWHNHR, respectively) were selected for subsequent analysis. Peptide C18, WPWHNHR, significantly inhibited HCVpp from entering Huh7.5 cells detected by flow cytometry. The HCVpp entry was notably inhibited in the presence of peptide C18, compared to the group of HCVpp and the group of the unrelated peptide. Furthermore, quantitative real-time RT-PCR demonstrated that the level of HCV RNA in HCVcc-infected Huh7.5 cells was significantly lower in the presence of peptide C18, compared to Huh7.5 cells incubated with HCVcc and the unrelated peptide. 2442 GWWYKGRARPVSAVA(2) WHWRHRIPLQLAAGR(2) GIGGVWYSSIVGPGR(2) SSPPRYTRSWHWHVR(1) RLLGAGRWWNVHPRV(1) SRWRGWHHSWLVVAN(1) FRWVPKFFSAAALPR(1) WRRWFYQFPTPLAAA(1) 8 Phage display (common panning) 3-4 PMID:17237992 The anticodon stem and loop of human tRNA(Lys3), ASL(Lys3) Not determined. Not determined. Not determined. f5-15mer phage display library (X15) Biotinylated unmodified ASL(Lys3) was attached to streptavidin selection plates. Three or four rounds of selection were conducted. Bound phages were eluted with acid elution buffer. 2443 APTTCSKPQWYCDNYV(2) 1 Phage display (common panning) 3-4 PMID:17237992 The anticodon stem and loop of human tRNA(Lys3), ASL(Lys3) Not determined. Not determined. Not determined. f88-4/Cys6 phage display library (X4CX6CX4) Biotinylated unmodified ASL(Lys3) was attached to streptavidin selection plates. Three or four rounds of selection were conducted. Bound phages were eluted with acid elution buffer. 2444 GIGGVWYSSIVGPGR(4) RVPPRYHAKISPMVK(2) GWWYKGRARPVSAVA(1) 3 Phage display (common panning) 3-4 PMID:17237992 The anticodon stem and loop of human tRNA(Lys3), ASL(Lys3) Not determined. Not determined. Not determined. f5-15mer phage display library (X15) Biotinylated unmodified ASL(Lys3) was attached to streptavidin selection plates. Three or four rounds of selection were conducted. Bound phages were eluted with alkaline elution buffer. 2445 WRRWFYQFPTPLAAA(16) RVPPRYHAKISPMVK(3) VRVYFGFGPPPYFGG(2) GLYLPLPSSMSVRAV(1) GRAIDGFPYFRTALL(1) 5 Phage display (common panning) 3-4 PMID:17237992 ASL(Lys3) (s2U34;Ψ39) Not determined. Not determined. Not determined. f5-15mer phage display library (X15) Biotinylated ASL(Lys3) (s2U34;Ψ39) was attached to streptavidin selection plates. Three or four rounds of selection were conducted. Bound phages were eluted with acid elution buffer. 2446 TRQQCQKWTLWCRTVL(2) TQEYCNAHNEKCFQRT(1) APTTCSKPQWYCDNYV(1) 3 Phage display (common panning) 3-4 PMID:17237992 ASL(Lys3) (s2U34;Ψ39) Not determined. Not determined. Not determined. f88-4/Cys6 phage display library (X4CX6CX4) Biotinylated ASL(Lys3) (s2U34;Ψ39) was attached to streptavidin selection plates. Three or four rounds of selection were conducted. Bound phages were eluted with acid elution buffer. 2447 RVPPRYHAKISPMLK(16) GIGGVWYSSIVGPGR(3) GWWYKGRARPVSAVA(1) VSVDGVTSVWHRLSG(1) PPKFPRTSSWRLSSL(1) WRRWFYQFPTPLAAA(1) 6 Phage display (common panning) 3-4 PMID:17237992 ASL(Lys3) (s2U34;Ψ39) Not determined. Not determined. Not determined. f5-15mer phage display library (X15) Biotinylated ASL(Lys3) (s2U34;Ψ39) was attached to streptavidin selection plates. Three or four rounds of selection were conducted. Bound phages were eluted with alkaline elution buffer. 2448 RVPPRYHAKISPMVK(11) WRRWFYQFPTPLAAA(6) RYASQLSDQILFTLP(2) GWWYKGRARPVSAVA(1) SSPPRYTRSWHWHVR(1) SRQTGHSRNSYTRLP(1) WPVPLAPYWSDLSPS(1) GSFALLRSAQASARP(1) 8 Phage display (common panning) 3-4 PMID:17237992 The anticodon stem and loop of human tRNA(Lys3), ASL(Lys3) Not determined. Not determined. Not determined. f5-15mer phage display library (X15) Biotinylated unmodified ASL(Lys3) was attached to streptavidin selection plates. Three or four rounds of selection were conducted. With Zn2+ present, bound phages were eluted with acid elution buffer. 2449 RVPPRYHAKISPMVK(3) WRRWFYQFPTPLAAA(3) 2 Phage display (common panning) 3-4 PMID:17237992 ASL(Lys3) (s2U34;Ψ39) Not determined. Not determined. Not determined. f5-15mer phage display library (X15) Biotinylated ASL(Lys3) (s2U34;Ψ39) was attached to streptavidin selection plates. Three or four rounds of selection were conducted. With Zn2+ present, bound phages were eluted with acid elution buffer. 2450 RERIHSPGSTQILF(2) PLLSLASHAQRTVGR(2) RSLWSDFYASASRGP(2) GWWYKGRARPVSAVA(1) GRAIDGFPYFRTALL(1) GEQGTNSRVSRLHTW(1) RGVFSHPHTAVPSHN(1) GWRFLVSSARVMRSD(1) GSRGGTLRGAEAGLP(1) WASHYMFHGYSVASD(1) LAVRRYALGQGYDWS(1) GVVAGASRPYVWWVS(1) AREYGTRFSLIGGYR(1) HSDVPACTSGDGCLA(1) GWRFLVSSARVMRSD(1) 15 Phage display (subtractive panning) 3-4 PMID:17237992 ASL(Lys3) (s2U34;Ψ39) Not determined. Not determined. Not determined. f5-15mer phage display library (X15) Biotinylated unmodified ASL(Lys3) and ASL(Lys3) (s2U34;Ψ39) were immobilized on the surface of streptavidin-coated microplates. The phage dsiplay library was pre-incubated with unmodified ASL(Lys3). The phages that were easily washed from the plate were subjected to selection on ASL(Lys3) (s2U34;Ψ39). Bound phages were eluted with acid elution buffer. 2451 FSVSFPSLPAPPDRS(18) GRVTYYSCGVSLLFQ(4) AGPVPLHSLSYYYNQ(1) 3 Phage display (subtractive panning) 4 PMID:21762809 ASL(Lys3) (UUU-ms2t6A37;mcm5s2U34;Ψ39) Not determined. Not determined. Not determined. f5-15mer phage display library (X15) Biotinylated unmodified hASL(Lys3) (UUU) and ASL(Lys3) (UUU-ms2t6A37;mcm5s2U34;Ψ39) were attached to streptavidin selection plates. The first round of screening was conducted with the unmodified hASL(Lys3) (UUU), followed by multiple rounds of screening with the triply modified hASL(Lys3)(UUU). Bound phages were eluted from the plates using basic condition. 2452 RVTHHAFLGAHRTVG(10) RAVMTVVWPVSFAGF(5) PAVASTSSLIIDGPF(2) PKAFQYGGRAVGGLW(1) AAHVSEHYVSGSLRP(1) ASVGPAPWAMTPPVS(1) APALWYPWRSLLPLY(1) ASLHPVPKTWFFLLS(1) WSHSRNTADVPVSML(1) 9 Phage display (subtractive panning) 4 PMID:21762809 ASL(Lys3) (UUU-ms2t6A37;mcm5s2U34;Ψ39) Not determined. Not determined. Not determined. f5-15mer phage display library (X15) Biotinylated unmodified hASL(Lys3) (UUU) and ASL(Lys3) (UUU-ms2t6A37;mcm5s2U34;Ψ39) were attached to streptavidin selection plates. The first round of screening was conducted with the unmodified hASL(Lys3) (UUU), followed by multiple rounds of screening with the triply modified hASL(Lys3)(UUU). Bound phages were eluted from the plates using acid condition. 2453 HRGYCRDRWNCGEYF(8) PHRQCSAPAKSCKILP(8) TLPACHELPKHCKRRG(4) TLPACHELPKHCNEAR(1) 4 Phage display (subtractive panning) 4 PMID:21762809 ASL(Lys3) (UUU-ms2t6A37;mcm5s2U34;Ψ39) Not determined. Not determined. Not determined. f88-4/Cys6 phage display library (X4CX6CX4) Biotinylated unmodified hASL(Lys3) (UUU) and ASL(Lys3) (UUU-ms2t6A37;mcm5s2U34;Ψ39) were attached to streptavidin selection plates. The first round of screening was conducted with the unmodified hASL(Lys3) (UUU), followed by multiple rounds of screening with the triply modified hASL(Lys3)(UUU). Bound phages were eluted from the plates using basic condition. 2454 NGPECNAYMVRCRGYH(4) GNSNCPMLNEQCPWQD(1) HTETCINIRNTCTTVA(1) LKLPCKITINNCQLAG(1) 4 Phage display (subtractive panning) 4 PMID:21762809 ASL(Lys3) (UUU-ms2t6A37;mcm5s2U34;Ψ39) Not determined. Not determined. Not determined. f88-4/Cys6 phage display library (X4CX6CX4) Biotinylated unmodified hASL(Lys3) (UUU) and ASL(Lys3) (UUU-ms2t6A37;mcm5s2U34;Ψ39) were attached to streptavidin selection plates. The first round of screening was conducted with the unmodified hASL(Lys3) (UUU), followed by multiple rounds of screening with the triply modified hASL(Lys3)(UUU). Bound phages were eluted from the plates using acid condition. 2455 AHSANNFDVKGI GHSHPTLGWTNG HIWTGSIWRTWD HLYHFSSWLYAR KFLLPNPQAGAP LSTASTEKSWIN MPLMSEPALEML QAHNPNRYMASW SSLSTYHHRWHL TPMVERNYNAAD YFDWGLSNANDT YSAHNYIGDSGP 12 Phage display (common panning) PMID:24266517 Anti-DENV E EDIII HuScFv Envelope protein E Not determined. Not determined. Ph.D.-12 phage display library (X12) 2456 VSSTQDFP(13)[26.032] DGSIPWST(8)[28.087] AFSEAAQT(5)[14.013] ADAPSNGW(1)[12.007] DNDNYAFP(1)[2.396] DYADYSDL(1)[ND] AADYDPLA(1)[ND] 7 Phage display (subtractive panning) 4 PMID:20735141 Human breast carcinoma SKBR-3 (HER positive) cells Not determined. Not determined. Not determined. f8-8mer phage display library (X8) ELISA Binding of wild type phage and affinity selected phages to SKBR-3 cells was measured by ELISA. The ELISA signal in milliabsorbance units (mAU)/min was determined, reproduced from the graph and shown. The value of wild type phage was 2.996. ND denotes not determined. The f8/8 landscape phage library was first incubated with empty culture flask to remove the phages that specifically bound to the culture flask. The resultant depleted library was then used as an input library to the culture flask containing the control MCF-10A cells (HER negative cells). This negative selection was performed to remove phage that binds to the receptors of healthy cells. Then the depleted phage library was transferred to the flask with target SKBR-3 cancer cells (HER positive cells). During each round of affinity selection, after cell-surface-bound phages were eluted, the cells were lysed to release the cell-internalized phages to obtain cell-internalizing peptides. The cell-internalized phages were recovered from cell lysates. 2457 DGSIPWST(13)[28.087] VSSTQDFP(10)[26.032] DNGSGISW(5)[24.026] DTKTAPAW(3)[16.117] GSTDSDLK(2)[12.007] ATTAPSYP(1)[2.043] AAATKSDL(1)[2.043] 7 Phage display (subtractive panning) 4 PMID:20735141 Human breast carcinoma SKBR-3 (HER positive) cells Not determined. Not determined. Not determined. f8-8mer phage display library (X8) ELISA Binding of wild type phage and affinity selected phages to SKBR-3 cells was measured by ELISA. The ELISA signal in milliabsorbance units (mAU)/min was determined, reproduced from the graph and shown. The value of wild type phage was 2.996. The f8/8 landscape phage library was first incubated with empty culture flask to remove the phages that specifically bound to the culture flask. The resultant depleted library was then used as an input library to the culture flask containing the control MCF-10A cells (HER negative cells). This negative selection was performed to remove phage that binds to the receptors of healthy cells. Then the depleted phage library was transferred to the flask with target SKBR-3 cancer cells (HER positive cells). During each round of affinity selection, cell-surface-bound phages were eluted. 2458 LTHPRWP(2) TTYNSPP(2) ARPLEIT(2) TDRLHFL(2) HPSRSLD(1) TFAKSAY(1) TYSTLGY(1) EQFSAPI(1) QHHPTYM(1) HSHYSLK(1) MSLSHIT(1) FTHPRRR(1) SMYGSYN(1) SLDALLS(1) TQLLEPT(1) ARPRNIT(1) 16 Phage display (subtractive panning) 2 PMID:24659568 Human synovium-derived mesenchymal stem cells, hSMSCs Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the negative selection procedure, the Ph.D.-7 phage display library was incubated with the fibroblasts for 1 h to exclude the fibroblast affinity phage clones. The supernatant was then collected and incubated with hSMSCs for 1 h to allow hSMSCs internalization. Subsequently, the hSMSCs were washed for 10 min to remove the poorly binding phage clones. The hSMSCs were neutralized with 150 mL of 1 M Tris-HCl (pH 9.1), and lysed in 20% NP-40 for 30 min. The phage clones from the lysed hSMSCs were collected and amplified in Escherichia coli ER2738 (NEB), and then titrated and purified. The identified peptide (LTHPRWP) was designated as an hSMSCs-affinity peptide (L7). PRHLPTW (P7), a randomly scrambled peptide, was used as the negative control, whereas RGD, a peptide consisting of three amino acids (arginine, glycine, and aspartic acid), was used as the positive control. The hSMSCs were incubated for 1 h with FITC-labeled L7, RGD, or P7, and measured via FCM. The average fluorescence intensity was 126.2 ± 13.0 for the hSMSCs incubated with FITC-L7, 118.1 ± 17.3 for the hSMSCs incubated with FITC-RGD, and 5.4 ± 2.2 for the hSMSCs incubated with FITC-P7. To investigate the species specificity of the identified peptide, rat and rabbit SMSCs were also cultured and incubated with FITC-L7, RGD, or P7, and the fluorescent intensities were measured via FCM. This result suggests that L7 has high affinity and specificity for SMSCs and can bind with human, rat, and rabbit SMSCs without species specificity. Thereafter, L7 was covalently conjugated onto both polycaprolactone (PCL) electrospun meshes and human decalcified bone scaffolds (hDBSc) to investigate its TE applications. After 24 h coculture with human SMSCs (hSMSCs), L7-conjugated PCL electrospun meshes had significantly more adherent hSMSCs than the control group, and the cells expanded well. Similar results were obtained using hDBSs. These results suggest that the novel L7 peptide sequence has a high specific affinity to SMSCs. Covalently conjugating this peptide to either artificial polymer material (PCL mesh) or natural material (hDBS) significantly enhances the adhesion of SMSCs. 2459 LTHPRWP(3/20) YSIPKSS(3/20) SILPYPY(2/20) TFAKSAY(2/20) ASLVRMM(2/20) IVLPYPI(1/20) HSHYSLK(1/20) DFKLPAS(1/20) SETATHP(1/20) SWHFVVS(1/20) EQFSAPI(1/20) VPPSMRP(1/20) 12 Phage display (subtractive panning) 3 PMID:24659568 Human synovium-derived mesenchymal stem cells, hSMSCs Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the negative selection procedure, the Ph.D.-7 phage display library was incubated with the fibroblasts for 1 h to exclude the fibroblast affinity phage clones. The supernatant was then collected and incubated with hSMSCs for 1 h to allow hSMSCs internalization. Subsequently, the hSMSCs were washed for 10 min to remove the poorly binding phage clones. The hSMSCs were neutralized with 150 mL of 1 M Tris-HCl (pH 9.1), and lysed in 20% NP-40 for 30 min. The phage clones from the lysed hSMSCs were collected and amplified in Escherichia coli ER2738 (NEB), and then titrated and purified. Twenty phage clones were randomly selected. An empty phage was found. The identified peptide (LTHPRWP) was designated as an hSMSCs-affinity peptide (L7). PRHLPTW (P7), a randomly scrambled peptide, was used as the negative control, whereas RGD, a peptide consisting of three amino acids (arginine, glycine, and aspartic acid), was used as the positive control. The hSMSCs were incubated for 1 h with FITC-labeled L7, RGD, or P7, and measured via FCM. The average fluorescence intensity was 126.2 ± 13.0 for the hSMSCs incubated with FITC-L7, 118.1 ± 17.3 for the hSMSCs incubated with FITC-RGD, and 5.4 ± 2.2 for the hSMSCs incubated with FITC-P7. To investigate the species specificity of the identified peptide, rat and rabbit SMSCs were also cultured and incubated with FITC-L7, RGD, or P7, and the fluorescent intensities were measured via FCM. This result suggests that L7 has high affinity and specificity for SMSCs and can bind with human, rat, and rabbit SMSCs without species specificity. Thereafter, L7 was covalently conjugated onto both polycaprolactone (PCL) electrospun meshes and human decalcified bone scaffolds (hDBSc) to investigate its TE applications. After 24 h coculture with human SMSCs (hSMSCs), L7-conjugated PCL electrospun meshes had significantly more adherent hSMSCs than the control group, and the cells expanded well. Similar results were obtained using hDBSs. These results suggest that the novel L7 peptide sequence has a high specific affinity to SMSCs. Covalently conjugating this peptide to either artificial polymer material (PCL mesh) or natural material (hDBS) significantly enhances the adhesion of SMSCs. 2460 LTHPRWP(11/20) SILPYPY(2/20) SHPASHD(1/20) TFAKSAY(1/20) TDRLHFL(1/20) VRPHTSS(1/20) SAWTYEY(1/20) DFKLPAS(1/20) 8 Phage display (subtractive panning) 4 PMID:24659568 Human synovium-derived mesenchymal stem cells, hSMSCs Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the negative selection procedure, the Ph.D.-7 phage display library was incubated with the fibroblasts for 1 h to exclude the fibroblast affinity phage clones. The supernatant was then collected and incubated with hSMSCs for 1 h to allow hSMSCs internalization. Subsequently, the hSMSCs were washed for 10 min to remove the poorly binding phage clones. The hSMSCs were neutralized with 150 mL of 1 M Tris-HCl (pH 9.1), and lysed in 20% NP-40 for 30 min. The phage clones from the lysed hSMSCs were collected and amplified in Escherichia coli ER2738 (NEB), and then titrated and purified. Twenty phage clones were randomly selected. An empty phage was found. The identified peptide (LTHPRWP) was designated as an hSMSCs-affinity peptide (L7). PRHLPTW (P7), a randomly scrambled peptide, was used as the negative control, whereas RGD, a peptide consisting of three amino acids (arginine, glycine, and aspartic acid), was used as the positive control. The hSMSCs were incubated for 1 h with FITC-labeled L7, RGD, or P7, and measured via FCM. The average fluorescence intensity was 126.2 ± 13.0 for the hSMSCs incubated with FITC-L7, 118.1 ± 17.3 for the hSMSCs incubated with FITC-RGD, and 5.4 ± 2.2 for the hSMSCs incubated with FITC-P7. To investigate the species specificity of the identified peptide, rat and rabbit SMSCs were also cultured and incubated with FITC-L7, RGD, or P7, and the fluorescent intensities were measured via FCM. This result suggests that L7 has high affinity and specificity for SMSCs and can bind with human, rat, and rabbit SMSCs without species specificity. Thereafter, L7 was covalently conjugated onto both polycaprolactone (PCL) electrospun meshes and human decalcified bone scaffolds (hDBSc) to investigate its TE applications. After 24 h coculture with human SMSCs (hSMSCs), L7-conjugated PCL electrospun meshes had significantly more adherent hSMSCs than the control group, and the cells expanded well. Similar results were obtained using hDBSs. These results suggest that the novel L7 peptide sequence has a high specific affinity to SMSCs. Covalently conjugating this peptide to either artificial polymer material (PCL mesh) or natural material (hDBS) significantly enhances the adhesion of SMSCs. 2461 CHLGYPGRC[0.656 ± 0.032] CHYSYPGVC[0.811 ± 0.045] CHLNYPGYC[0.846 ± 0.152] CHLRYPGEC[1.087 ± 0.105] CYKGYPGYC[0.293 ± 0.009] YPG 5 Phage display (common panning) 3 PMID:24497417 Anti-Claudin 18.2 monoclonal antibody IMAB362 Claudin 18.2 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Binding of picked clones to IMAB362 was detected by Phage ELISA. Absorbance at 450 nm was determined. Data shown were reproduced from the graph and expressed as the mean of two experiments ± S.D. Three of the peptide sequences (CHLGYPGRC, CHYSYPGVC and CHLNYPGYC) were selected for further binding, SAR analysis, and optimization via peptide microarrays. 2462 PRTGTWRSSIAYGGG(17) PGTGTWRSYLRFGGG(1) PVTGTWTSSIASWMG(1) PYTGTWRSSIWVLSG(1) PGTGTWRSWLSFNVG(1) 5 Phage display (common panning) 4 PMID:24356963 Fc region of human immunoglobulin G (IgG) Not determined. Not determined. Not determined. T7 PXTGTWX8G phage display library The phage library was biopanned against a biotinylated human Fc region (Jackson ImmunoResearch) immobilized on streptavidin MagneSphere@ paramagnetic particles (Promega). 2463 SWDIAWPPLKVP(6)[0.752 ± 0.029] NILPYNKTMLVK(3)[ND] SSSNTTTKHVFT(1)[ND] DSYYTKTERNTH(1)[ND] YEFPRSWDMETN(1)[ND] NRPDSAQFWLHH(1)[ND] GWEVTWPASYAF(1)[ND] 7 Phage display (subtractive panning) 4 PMID:24604160 Human glioblastoma cell line A172 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Specific binding of phage SWDI (displaying SWDIAWPPLKVP) to A172 was evaluated by phage ELISA. Absorbance at 405 nm was determined. The ASNL phage, the random phage from the phage libraries, was used as the control phage. The results represent the mean values ± SEM (standard error of the mean) of two independent experiments analyzed in triplicate. The value shown was reproduced from the graph. The value of the control phage was 0.202 ± 0.024. ND denotes not determined. The human embryonic kidney cell line HEK 293 was used for substraction; the human glioblastoma cell line A172 for selection. The entire subtraction/selection cycle was repeated four times. Immunofluorescence staining with anti-M13 phage antibody was performed to observe the specific binding of phage SWDI (displaying SWDIAWPPLKVP) to A172 cells. SWDI phages revealed by the red fluorescent foci were seen on the cell surface of A172; however, no fluorescent signals were present when A172 cells were incubated with the negative control, phage ASNL, which was picked randomly from phage library. Besides, results of ELISA shown that the binding activity of the phage displaying SWDIAWPPLKVP peptide in A172 was more than 10-fold higher than that of the control phage. Then the selected peptide SWDIAWPPLKVP was inserted into adenoviral hexon protein. As a result, the modified Ad5 had increased infectivity in A172 cells, compared with that in control cell lines. 2464 SRPKPPNPS(15)[4.116] CKRDSISPYSC(6)[9.892] KTKKPPNPS(4)[5.309] RLKPPNPTE(2)[4.216] RKPPNPPPP(1)[4.380] RRDTISPYS(1)[NT] KPPNP 6 Phage display (common panning) 1 PMID:8855309 IgG from cerebro spinal fluid of multiple sclerosis (MS) patient 1 Not determined. Not determined. Not determined. pVIII-9aa and pVIII-9aa.Cys phage display library pool ELISA Average values (A405) from two experiments have been determined. Results are expressed as the ratio between the average value of the tested clone and that of wild-type phage (clone/wt A405). Data shown were reproduced from the graph. NT denotes not tested. For each selection, CSF IgGs were immobilized using magnetic beads coated with goat anti-human IgG Fc-specific Ab. 2465 KPKTNQIRP(2)[NT] KKTGNITPK(1)[6.091] 2 Phage display (common panning) 1 PMID:8855309 IgG from cerebro spinal fluid of multiple sclerosis (MS) patient 2 Not determined. Not determined. Not determined. pVIII-9aa and pVIII-9aa.Cys phage display library pool ELISA Average values (A405) from two experiments have been determined. Results are expressed as the ratio between the average value of the tested clone and that of wild-type phage (clone/wt A405). Data shown were reproduced from the graph. NT denotes not tested. For each selection, CSF IgGs were immobilized using magnetic beads coated with goat anti-human IgG Fc-specific Ab. 2466 CKRDSISPYSC(6)[10.007, 1.035, 1.234] CYSATAGPSPC(2)[NT, NT, NT] CYKSTAGPWGC(1)[NT, NT, NT] CYGASDGPNAC(1)[NT, NT, NT] CKRDSIPPTSC(1)[NT, NT, NT] 5 Phage display (common panning) 1 PMID:9532590 IgG from the cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients Not determined. Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA Binding of the selected phagotopes to the CSF1, CSF2 and CSF3 was detected by ELISA on immobilized phage. Average values (A405nm) from two independent experiments have been determined. Results are expressed as the ratio between the average value of the tested phagotope and that of wild type phage (clone/wt A405 nm). Data shown were reproduced from the graph. NT denotes not tested. For each selection, 5 microgram of CSF IgG were immobilized using magnetic beads coated with goat anti-human IgG Fc-specific Ab. 2467 FDRLSLADVEQVSIEEAKAWALVQLRDW(11)[2.742, 3.362] WRTDVRWLSEFEEAEVEGKRPGVWLYGT(9)[6.704, 7.684] SSRADDVVWVSTGEEAAWRRQWLDLRTV(9)[2.339, 4.168] ADAVLPWGSSARWDIGGWQVASGGGTQA(5)[24.903, 26.285] GGGSPVRAEWAQDIRSRSEREMLVWLGK(2)[20.854, 23.347] AEPGVRERTEMEMGEIFGNLDLCMARRW(1)[13.420, 13.735] 6 Phage display (common panning) 1 PMID:9762658 IgG from cerebro spinal fluid of multiple sclerosis (MS) patient 1 Not determined. Not determined. Not determined. X28 and X28 M13 phage display library pool ELISA Binding of the selected phagotopes to the CSF and serum IgG from patient 1 was detected by ELISA. For each sample equal amounts of IgG were used and antibody recognition of the tested phagotope and wild type phage was measured. Average values (A405 nm) from two independent experiments have been determined. The background signal obtained on the wild type phage was subtracted. Results are expressed as the ratio between the subtracted value of the tested phagotope obtained by using CSF IgG and that observed using serum IgG (CSF A 405 nm/serum A 405 nm). These data shown were reproduced from the graph. Data obtained by using samples from the same patient at 2 years time interval are also shown. For each selection, CSF IgGs were immobilized using magnetic beads coated with goat anti-human IgG Fc-specific Ab. 2468 VPAWRSGGVGTYALLGCDPRISRCTNFS(7)[0.434] GLWGPPASSRSRPFPPSASVQRQQDVMM(4)[0.176] SGERARWDWSVFDRVDGLNQGEARIRIG(1)[0.324] FHPAEPRSRPLADREMCDVRLAVGGVVV(1)[0.166] 4 Phage display (common panning) 1 PMID:9762658 IgG from a mixture of 20 different CSF of multiple sclerosis (MS) patients Not determined. Not determined. Not determined. X28 and X28 M13 phage display library pool ELISA Binding of IgG from mixtures of 20 MS CSF to crude phage-containing supernatant was detected by ELISA on immobilized phage. For each sample absorbance (405 nm) was measured in two experiments with the phagotope and in two experiments with wild type phage. Data shown were reproduced from the graph. For each selection, CSF IgGs were immobilized using magnetic beads coated with goat anti-human IgG Fc-specific Ab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hage display (common panning) 3/4 PMID:10662687 Tyrosyl-tRNA synthetase, TyrRS Not determined. Not determined. Not determined. Twelve different M13 phage libraries pool ELISA Absorbance at 405 nm was measured. NT denotes not tested. 2470 SRDWGFWRLPESMA[0.83] SRDWGFWRLPNPTE[0.76] SRDWGFWDWGVDR[0.83] SREWHFWRDYNPT[0.85] SRDWSFWDVRDWA[0.70] SREWGFFDPRFLP[0.74] SRDWHMFFTGPGQ[0.74] SRDWHYFHGFPSV[0.78] SRDWALWHPPLSL[0.74] SRDWGYWHLGEIV[0.57] SRDWGYWKVPAPS[0.70] SREWGFFRMPLHD[0.70] SRDWGFWNMPPGS[0.58] SRDWGFWNPIGPL[0.80] SRDWAFWRGDASG[0.47] SREWAFFRNWDSV[0.70] SRCGWAAEVRGVC[0.70] SSCHWGREVRGLC[0.58] SSERGSGDRGEKG[0.55] R-[DE]-W-[GA]-[FMY]-[WF]-[DHKNR], C-x-W-x(2)-E-V-R-G-[VL]-C 19 Phage display (common panning) 3/4 PMID:10662687 Proline--tRNA ligase Not determined. Not determined. Not determined. Twelve different M13 phage libraries pool ELISA Absorbance at 405 nm was measured. Biotinylated-target protein and phage were incubated together in solution prior to capturing the target-phage complex on streptavidin-coated paramagnetic particles. Or the target protein was biotinylated and captured on streptavidin-coated 96-well plates. 2471 SSWQGNVLLGNWIS[0.60] SRPMWMNLVNYVS[1.07] SSVLCRGFCWTDT[0.89] SSWQCRGFCWAGE[0.68] SSPTCRGFCWSTS[0.20] SSNNAFCRFGCWQ[0.60] SSAGTTRHEELIP[0.20] MPPPEPR[0.14] 8 Phage display (common panning) 3 PMID:10662687 Alcohol dehydrogenase 2 Not determined. Not determined. Not determined. Twelve different M13 phage libraries pool ELISA Absorbance at 405 nm was measured. For the first round of panning, biotinylated-target protein was bound to streptavidin paramagnetic particles. For the second round of panning, biotinylated-target protein and phage were incubated together in solution prior to capturing the target-phage complex on streptavidin-coated paramagnetic particles. For the third round of panning, the target protein was biotinylated and captured on streptavidin-coated 96-well plates. 2472 SSQTDWRKIFQSL[0.36] SSSTDWLNVWRQL[0.15] SSATDWGRVYSIL[0.94] SSASYAPWPIYFA[0.76] SSGAFKPWPVYSF[0.36] SRQVEVFKPWPVY[0.44] SSSFKPWPIYLGS[0.52] SSEPFSVWPIYKH[0.44] SSSVPFAPWPVYA[0.62] SSTSLPFNRWPIY[0.22] T-D-W-x(2)-[IV]-[FWY]-x-L, [FY]-K-P-W-P-[IV]-Y 10 Phage display (common panning) 3/4 PMID:10662687 β-glucosidase Not determined. Not determined. Not determined. Twelve different M13 phage libraries pool ELISA Absorbance at 405 nm was measured. Biotinylated-target protein and phage were incubated together in solution prior to capturing the target-phage complex on streptavidin-coated paramagnetic particles. Or the target protein was biotinylated and captured on streptavidin-coated 96-well plates. 2473 SRYDNWGWTEKLTK[0.64] SSMYDEDQWILKLN[0.48] SSMFDDPSWTMLMR[0.61] SRYDDWGWVHRLS[0.50] SSYDDWDWVVRLN[0.66] SRWEWQEFLDGPL[0.55] SSVRWWSDDEWRM[0.83] [YF]-D-[DE]-W-x-W-x(2)-[KR]-[LM] 7 Phage display (common panning) 3/4 PMID:10662687 Hexokinase-1 and hexokinase-2 Not determined. Not determined. Not determined. Twelve different M13 phage libraries pool ELISA Absorbance at 405 nm was measured. The target protein was biotinylated and captured on streptavidin-coated 96-well plates. 2474 SSEEGRKFRWGWL[0.62] SSLPEERKLRWGWL[0.77] SSEDNRKLRWGWL[0.58] SSDARTCRWCWLG[0.44] SSWFMDCEWQCFT[0.15] SRWCGEHDRFCRQA[0.38] SRTRFCSHDDAFC[0.92] [ED]-x-R-K-[CFL]-R-W-[CG]-W-L, C-x-W-x-C, [FW]-C-x(3)-D-x-F-C 7 Phage display (common panning) 3/4 PMID:10662687 Glycogen phosphorylase, muscle form Not determined. Not determined. Not determined. Twelve different M13 phage libraries pool ELISA Absorbance at 405 nm was measured. The target protein was biotinylated and captured on streptavidin-coated 96-well plates. 2475 SRLLEVSPGWWQM[0.12] SSFRELKPGWWSY[0.09] SSWGDYFNWRDGL[0.09] E-[VL]-x-P-G-W-W-x-[MY] 3 Phage display (common panning) 3/4 PMID:10662687 Carboxypeptidase B Not determined. Not determined. Not determined. Twelve different M13 phage libraries pool ELISA Absorbance at 405 nm was measured. The target protein was biotinylated and captured on streptavidin-coated 96-well plates. 2476 RGLFTEWFRGGSWSNYRVTS 1 Phage display (common panning) 3 PMID:11602024 MRNA of amyloid precursor protein (APP) Not determined. Not determined. Not determined. X20 phage display library For the mRNA pans, all solutions and surfaces are pretreated with DEPC or RNaseZap, respectively, to eliminate RNase contamination that may compromise the integrity of the RNA. The biotinylated RNA solution was added to an appropriate number of wells in a 96-well microtiter plate precoated with Streptavidin (Pierce). 2477 TDGGRSVISDNVRGGSRLWLWIRHGSWSQAWGPQDAWSSK RVSSAQPGCTSRVRFRCPRGGLLFNGVTSTNPKTGLSNAQ 2 Phage display (common panning) 3 PMID:11602024 MRNA of amyloid precursor protein (APP) Not determined. Not determined. Not determined. X40 phage display library For the mRNA pans, all solutions and surfaces are pretreated with DEPC or RNaseZap, respectively, to eliminate RNase contamination that may compromise the integrity of the RNA. The biotinylated RNA solution was added to an appropriate number of wells in a 96-well microtiter plate precoated with Streptavidin (Pierce). 2478 WPPGRTLSDLIRGGAGARGM SSGGLHRWSALRGGHGHGLA 2 Phage display (common panning) 3 PMID:11602024 MRNA of hepatitis C virus (HCV) Not determined. Not determined. Not determined. X20 phage display library For the mRNA pans, all solutions and surfaces are pretreated with DEPC or RNaseZap, respectively, to eliminate RNase contamination that may compromise the integrity of the RNA. The biotinylated RNA solution was added to an appropriate number of wells in a 96-well microtiter plate precoated with Streptavidin (Pierce). 2479 AMRLKPIAFKGPRAGAGWVEVQPCFAAFRAACTRGGSHHH LHAGWDVTAPRRACKGAQGPGLHGRFYCHRGGLCSGLGRC DEQSSLKGKLRGALVRLGMGHAMPHRGGVWPSTGRPSKQG WTPRHGPMRCWRHQSVFPVGAGPHWALWPIKGPRGGRTAC PKTGSNIWLPLYHKVCPASTRAGNGRGGSRFLWGSMQTNC 5 Phage display (common panning) 3 PMID:11602024 MRNA of hepatitis C virus (HCV) Not determined. Not determined. Not determined. X40 phage display library For the mRNA pans, all solutions and surfaces are pretreated with DEPC or RNaseZap, respectively, to eliminate RNase contamination that may compromise the integrity of the RNA. The biotinylated RNA solution was added to an appropriate number of wells in a 96-well microtiter plate precoated with Streptavidin (Pierce). 2480 RLQRRGGGAVAVVWVGFGVGLLWGRLLLIILGWVLMWFLS QHSEHGGTEWRKRGGMAFAASFLCMRDSYRTTRLRSLLG GTRHVINRVRDSSGVPCKRFGGLQFSQMGKCTIPRGGA 3 Phage display (common panning) 3 PMID:11602024 MRNA of insulin-like growth factor-1 (IGF-1) Not determined. Not determined. Not determined. X40 phage display library For the mRNA pans, all solutions and surfaces are pretreated with DEPC or RNaseZap, respectively, to eliminate RNase contamination that may compromise the integrity of the RNA. The biotinylated RNA solution was added to an appropriate number of wells in a 96-well microtiter plate precoated with Streptavidin (Pierce). 2481 VLRGGSVGKGSLMWCQEVDWRTGGPRSNLWGLWNGRQPPK 1 Phage display (common panning) 3 PMID:11602024 MRNA of Rev Response Element (RRE) Not determined. Not determined. Not determined. X40 phage display library For the mRNA pans, all solutions and surfaces are pretreated with DEPC or RNaseZap, respectively, to eliminate RNase contamination that may compromise the integrity of the RNA. The biotinylated RNA solution was added to an appropriate number of wells in a 96-well microtiter plate precoated with Streptavidin (Pierce). 2482 VVYVGVLSYWPHLSGGGRLQVRCLIGRGGFGCRGG 1 Phage display (common panning) 4 PMID:11602024 MRNA of amyloid precursor protein (APP) Not determined. Not determined. Not determined. X40 phage display library For the mRNA pans, all solutions and surfaces are pretreated with DEPC or RNaseZap, respectively, to eliminate RNase contamination that may compromise the integrity of the RNA. The biotinylated RNA solution was added to an appropriate number of wells in a 96-well microtiter plate precoated with Streptavidin (Pierce). 2483 CRGRRST CRSRKG CKAAKNK CKGAKAR FRVGVADV 5 Phage display (in vivo) 2 PMID:14667506 RIP1-Tag2 tumors Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The screening process involved two ex vivo selection rounds followed by 2 in vivo selection rounds. Fluorescence detection assay showed that no peptides homed appreciably to normal pancreatic islets or other organs. Besides, The relative homing efficiencies in the various tumor models of the phage from the RIP1-Tag2 tumor screen fall broadly into two categories: those that selectively home to RIP1-Tag2 tumors (CKAAKNK, CRGRRST, FRVGVADV) and those that show a more general homing to other tumors in addition to RIP1-Tag2 (VGVG, CKGAKAR). The phage homing data were supported by i.v. injection of fluorescein-conjugated peptides corresponding to the phage. Peptide CRGRRST is homologous with pro-PDGF-B, which is expressed in endothelial cells, while its receptor is expressed in pericytes. 2484 CEYQLDVE 1 Phage display (in vivo) 3 PMID:14667506 RIP1-Tag2 angiogenic islets Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The screening process involved two ex vivo selection rounds followed by 3 in vivo selection rounds. Fluorescence detection assay showed that the peptide CEYQLDVE didn't home appreciably to normal pancreatic islets or other organs. 2485 CWDDGWLC(131) CWDDLWWLC(10) CWDDGLMC(7) CWDDGWMC(4) CWDD[GL]WLC 4 Phage display (competitive panning) 4 PMID:7657703 Fibronectin type Ⅲ10(RGD)-containing fragments Not determined. Not determined. Not determined. CX5C+CX6C+CX7C+CX9 phage display library pool After the fourth panning, wells with bound phages were eluted with 1 mM solution of GRGDSP or *CELRGDGWC* peptidee. A predominant cyclic motif, *CWDD[GL]WLC*, was obtained (the asterisks denote a potential disulfide bond). Studies using the purified phage and the corresponding synthetic cyclic peptides showed that *CWDDGWLC*-expressing phage binds specifically to fibronectin and to fibronectin fragments containing the RGD sequence. The binding did not require divalent cations and was inhibited by both RGD and *CWDDGWLC*-containing synthetic peptides. Conversely, RGD-expressing phage attached specifically to immobilized *CWDDGWLC*-peptide and the binding could be blocked by the respective synthetic peptides in solution. Moreover, fibronectin bound to a *CWDDGWLC*-peptide affinity column, and could be eluted with an RGD-containing peptide. The *CWDDGWLC*-peptide inhibited RGD-dependent cell attachment to fibronectin and vitronectin, but not to collagen.A region of the β3 subunit of RGD-binding integrins that has been previously demonstrated to be involved in ligand binding includes a polypeptide stretch, KDDLW (in β3) similar to WDDC/LWL. Synthetic peptides corresponding to this region in β3 were found to bind RGD-displaying phage and conversion of its two aspartic residues into alanines greatly reduced the RGD binding. Polyclonal antibodies raised against the *CWDDGWLC*-peptide recognized β1 and β3 in immunoblots. These data indicate that the *CWDDGWLC*-peptide is a functional mimic of ligand binding sites of RGD-directed integrins, and that the structurally similar site in the integrin β subunit is a binding site for RGD. 2486 CWDDGWLC(21) CLVWLLVQFY(2) CTFGGGIGRV(l) CTLRFQRSC(1) CWDD[GL]WLC 4 Phage display (common panning) 4 PMID:7657703 Fibronectin type Ⅲ10(RGD)-containing fragments Not determined. Not determined. Not determined. CX5C+CX6C+CX7C+CX9 phage display library pool After the fourth panning, wells with bound phages were eluted with 2 mM EDTA. A predominant cyclic motif, *CWDD[GL]WLC*, was obtained (the asterisks denote a potential disulfide bond). Studies using the purified phage and the corresponding synthetic cyclic peptides showed that *CWDDGWLC*-expressing phage binds specifically to fibronectin and to fibronectin fragments containing the RGD sequence. The binding did not require divalent cations and was inhibited by both RGD and *CWDDGWLC*-containing synthetic peptides. Conversely, RGD-expressing phage attached specifically to immobilized *CWDDGWLC*-peptide and the binding could be blocked by the respective synthetic peptides in solution. Moreover, fibronectin bound to a *CWDDGWLC*-peptide affinity column, and could be eluted with an RGD-containing peptide. The *CWDDGWLC*-peptide inhibited RGD-dependent cell attachment to fibronectin and vitronectin, but not to collagen.A region of the β3 subunit of RGD-binding integrins that has been previously demonstrated to be involved in ligand binding includes a polypeptide stretch, KDDLW (in β3) similar to WDDC/LWL. Synthetic peptides corresponding to this region in β3 were found to bind RGD-displaying phage and conversion of its two aspartic residues into alanines greatly reduced the RGD binding. Polyclonal antibodies raised against the *CWDDGWLC*-peptide recognized β1 and β3 in immunoblots. These data indicate that the *CWDDGWLC*-peptide is a functional mimic of ligand binding sites of RGD-directed integrins, and that the structurally similar site in the integrin β subunit is a binding site for RGD. 2487 CWDDGWLC(45) CSWDDGWLC(6) CPDDLWWLC(3) CDGWLGFC(2) CQRIVLGFT(1) CDYWLGFC(1) CFVLWLVC(1) CGNRLRC(1) CWDD[GL]WLC 8 Phage display (common panning) 4 PMID:7657703 Fibronectin type Ⅲ10(RGD)-containing fragments Not determined. Not determined. Not determined. CX5C+CX6C+CX7C+CX9 phage display library pool After the fourth panning, wells with bound phages were directly incubated with 50 μl of bacteria. A predominant cyclic motif, *CWDD[GL]WLC*, was obtained (the asterisks denote a potential disulfide bond). Studies using the purified phage and the corresponding synthetic cyclic peptides showed that *CWDDGWLC*-expressing phage binds specifically to fibronectin and to fibronectin fragments containing the RGD sequence. The binding did not require divalent cations and was inhibited by both RGD and *CWDDGWLC*-containing synthetic peptides. Conversely, RGD-expressing phage attached specifically to immobilized *CWDDGWLC*-peptide and the binding could be blocked by the respective synthetic peptides in solution. Moreover, fibronectin bound to a *CWDDGWLC*-peptide affinity column, and could be eluted with an RGD-containing peptide. The *CWDDGWLC*-peptide inhibited RGD-dependent cell attachment to fibronectin and vitronectin, but not to collagen.A region of the β3 subunit of RGD-binding integrins that has been previously demonstrated to be involved in ligand binding includes a polypeptide stretch, KDDLW (in β3) similar to WDDC/LWL. Synthetic peptides corresponding to this region in β3 were found to bind RGD-displaying phage and conversion of its two aspartic residues into alanines greatly reduced the RGD binding. Polyclonal antibodies raised against the *CWDDGWLC*-peptide recognized β1 and β3 in immunoblots. These data indicate that the *CWDDGWLC*-peptide is a functional mimic of ligand binding sites of RGD-directed integrins, and that the structurally similar site in the integrin β subunit is a binding site for RGD. 2488 SVSVGMKPSPRP(3) ASPMPTAKLRFA(1) NWTTNRPNNTSP(1) MNNDHVPSVAQG(1) AYWSNPESHPXQ(1) SNPFSKPYGLTV(1) WDSNTYTPRPLM(1) ATNPTHPGHLPV(1) NPSLHKPLTISI(1) NPYQHRNWAYVG(1) HARENSLPWLHA(1) YPHYSLPGSSTL(1) LPLALPRHNASV(1) SSLKLHQTPLDA(1) SSLEPWHRTTSR(1) GSIALSSWLSLP(1) GNFQSESYFVPH(1) GLHESTFTQRRL(1) FSHELSWKPRKA(1) NYLHNHPYGTVG(1) LNYFTLSSKRE(1) 21 Phage display (in vivo) 5 PMID:19276080 SAS-derived tumors Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) SAS cells were injected subcutaneously into the dorsolateral flank of 4–6-week-old SCID mice to produce oral cancer xenografts. The phage-displayed peptide library was injected intravenously into the tail vein of SCID mice bearing size-matched SAS-derived tumors. From 30 random selected phage clones, 23 phage-displayed peptide sequences were identified. Six novel peptides (NYLHNHPYGTVG, SNPFSKPYGLTV, GLHESTFTQRRL, YPHYSLPGSSTL, SSLEPWHRTTSR and LPLALPRHNASV) were identified as being able to recognize tumor vasculature but not normal blood vessels in severe combined immunodeficiency (SCID) mice bearing human tumors. These tumor-homing peptides also bound to blood vessels in surgical specimens of various human cancers. The peptidelinked liposomes containing fluorescent substance were capable of translocating across the plasma membrane through endocytosis. With the conjugation of peptides and liposomal doxorubicin, the targeted drug delivery systems enhanced the therapeutic efficacy of the chemotherapeutic agent against human cancer xenografts by decreasing tumor angiogenesis and increasing cancer cell apoptosis. Furthermore, the peptide-mediated targeting liposomes improved the pharmacokinetics and pharmacodynamics of the drug they delivered compared with nontargeting liposomes or free drugs. It indicates that the tumor-homing peptides can be used specifically target tumor vasculature and have the potential to improve the systemic treatment of patients with solid tumors. 2489 IVWHRWYAWSPASRI[581.1 ± 98.0] YYAWHWYAWSPKSV[69.9 ± 19.6] 2 Phage display (common panning) 3 PMID:12760424 Thomsen-Friedenreich antigen (T antigen, TF antigen) Not determined. Not determined. Not determined. fUSE5 X5WYA[WF]SPX4 phage dsiplay library Fluorescence Binding of the peptides to the TF antigen displayed on ASF was detected by observing a quenching of AlexaFluor 488 fluorescence. The dissociation constant (Kd, nM) that described binding affinity was calculated. The value of the first-generation peptide, P30 (HGRFILPWWYAFSPS), was 321.9 ± 70.0. And the control peptide, RNVPPIFNDVYWIAF, showed no detectable binding. Thomsen-Friedenreich (TF) antigen conjugated to human serum albumin (HSA) was immobilized on a streptavidin-coated Petri dish. 2490 ADITDPMGA(2) AVSPNVHDG(2) AAPDLQDAM(1) ADRLNSDAG(1) ADRPSTTSL(1) ADPPRTVST(1) ADRPSMSPT(1) ADRTSNAST(1) ADKSYIPSS(1) AVRNPSHHS(1) ADPTPRGHS(1) ADPTRQPHS(1) AEHQNSAGP(1) ADARSAGAIS(1) ADSKNAGPM(1) AETKFSGSA(1) ADPKGSGVT(1) AGLTSPNDM(1) AVGTHTPDS(1) 19 Phage display (common panning) PMID:12089009 Phytophthora capsici zoospore Not determined. Not determined. Not determined. f8-8mer phage display library (X8) In the affinity selection procedure, P. capsici zoospores were added to phage in the presence of 50 mM LiCl, which prevents zoospore encystment. The abilities of phage clones to induce encystment varied. For example, at a concentration of 2.5e9 virions/μl, clones Pc78 (displaying ADPPRTVST), Pc87 (displaying ADRPSMSPT), Pc64 (displaying ADITDPMGA), and Pc42 (displaying AVSPNVHDG) induced >80% encystment within 20 min. Conversely, only about 20% of zoospores exposed to Pc56 (displaying AAPDLQDAM), Pc45 (displaying AEHQNSAGP), Pc15B (displaying AGLTSPNDM), or the fuse5 phage vector were encysted during this period. In solution, 200 μM peptide 87, corresponding to phage Pc87, induced complete encystment of zoospores. In contrast, peptide 56, corresponding to the ineffectual phage Pc56, did not induce significant encystment at the same concentration. Peptide 42, corresponding to phage Pc42, induced an intermediate level of encystment. 2491 MTHDPVISLPTT LHRHSHGHSYKS GHWHNHRHQAPL KYPHQHLHMHDS IGWHYYLRTQHS YPFHHKHWHRPN ANIVPIHANHFQ MHKHPHGSQGPT YKLPGHHHHYRP 9 Phage display (common panning) PMID:12795832 CD147 Metuximab Not determined. Not determined. Ph.D.-12 phage display library (X12) 2492 GNVPDMRHIARP[420 ± 30] LTNGPHIPPDHP[430 ± 50] DYQHATKTGPHL[NT] APIMIQCPTPHT[NT] PYTHAATPPHSP[NT] SPPDFXMRPPHG[NT] VPPKPSRPASMW[NT] XPPYAVLAPTAL[NT] SAHEPPGSQRVA[NT] 9 Phage display (subtractive panning, competitive panning) 4 PMID:17132933 HIV-1 TAR Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay The association of the purified peptides to TAR was determined by filter binding assay. The dissociation constant KD (nM) value was calculated. The library was counter-selected by two successive incubations in the presence of streptavidin coatd beads. Nonbinding phages were recovered and incubated with 3' biotinylated TAR. The mix was then added to streptavidin beads. In the subsequent rounds, competitor yeast tRNA was added. No magnesium was used in the first selection round, but magnesium was used in subsequent selection rounds. 2493 LTNGPHIPPDHP[430 ± 50] 1 Phage display (subtractive panning, competitive panning) 4 PMID:17132933 HIV-1 TAR Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay The association of the purified peptides to TAR was determined by filter binding assay. The dissociation constant KD (nM) value was calculated. The library was counter-selected by two successive incubations in the presence of streptavidin coatd beads. Nonbinding phages were recovered and incubated with 3' biotinylated TAR. The mix was then added to streptavidin beads. In the subsequent rounds, competitor yeast tRNA was added. 2494 KVSSPPSPPHIL[550 ± 50] 1 Phage display (subtractive panning, competitive panning) 3 PMID:17132933 HIV-1 TAR Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay The association of the purified peptides to TAR was determined by filter binding assay. The dissociation constant KD (nM) value was calculated. The library was counter-selected by two successive incubations in the presence of streptavidin coatd beads. Nonbinding phages were recovered and incubated with 3' biotinylated TAR. The mix was then added to streptavidin beads. In the subsequent rounds, competitor yeast tRNA was added. 2495 GNVPDMRHIARP[420 ± 30] IXAKDQNTWPRR[NT] VEWPAQRHHHIP[NT] YASKFPHLXTLE[NT] HLSLPQAPTLLP[NT] ADLPSLFASRGT[NT] 6 Phage display (subtractive panning, competitive panning) 2 PMID:17132933 HIV-1 TAR Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay The association of the purified peptides to TAR was determined by filter binding assay. The dissociation constant KD (nM) value was calculated. NT denotes not tested. The library was counter-selected by two successive incubations in the presence of streptavidin coatd beads. Nonbinding phages were recovered and incubated with 3' biotinylated TAR. The mix was then added to streptavidin beads. In the subsequent rounds, competitor yeast tRNA was added. 2496 CGLIIQKNEC(3/24) CNAGESSKNC(2/24) 2 Phage display (competitive panning) 3 PMID:16476999 Human plasma clots Not determined. Not determined. Not determined. CX8C T7 phage display library Human plasma clots were incubated with the CX8C peptide library and extensively washed with PBS. Plasma was then added to the clot to remove phage recognizing soluble plasma components. When injected intravenously into mice bearing various types of tumors, fluorescein-conjugated CLT peptides (CGLIIQKNEC and CNAGESSKNC) accumulated in a fibrillar meshwork in the extracellular compartment of the tumors, but were not detectable in other tissues of the tumor-bearing mice. The tumor homing of both peptides was strongly reduced after coinjection with unlabeled CLT2 (CNAGESSKNC), indicating that the two peptides recognize the same binding site. The CLT peptide fluorescence colocalized with staining for fibrin(ogen) present in the extravascular compartment of tumors, but not in other tissues. The CLT peptides did not home to tumors grown in fibrinogen-null mice or in mice that lack plasma fibronectin. The CLT peptides also accumulated at the sites of injury in arteries, skeletal muscle, and skin. It is concluded that the CLT peptides recognize fibrin–fibronectin complexes formed by clotting of plasma proteins that have leaked into the extravascular space in tumors and other lesions. These peptides may be useful in targeting diagnostic and therapeutic materials into tumors and injured tissues. 2497 CHWSFGIFC(8) CHWMFSPWC(3) CPWYFLPWC(2) CPWYFQPWC(2) CPWYFHPYC(1) CPWYFFDLC(1) CPWYFFGAC(1) CSWIFWEYC(1) CSWIFWQYC(1) CSWIFWVYC(1) x-W-x-F-x(3) 10 Phage display (competitive panning) 3-5 PMID:9094664 Puumala virus Not determined. Not determined. Not determined. CX7C phage display library Biopanning with MAb 4G2 as the eluting antibody included three antibody and two acid elution rounds. In the first panning, the wells were rinsed 10 times with 5% NFDMP-TBS, pre-eluted with antibody 5A2, rinsed three times, and eluted with MAb 4G2. The second panning was done directly with MAb 4G2, whereas in the third, MAb3H9 was used as a pre-eluting MAb. For removal of residual antibody, the phage amplification product of the third panning was treated with Prosep-A (Bioprocessing Ltd.). The fourth panning was done conventionally with 0.1 M HCl-glycine, pH2.2, containing 1mg of BSA per ml as the eluent. The peptide insert, CHWMFSPWC, when displayed on the phages, completely inhibited Puumala virus infection in cell culture at the same effective concentration as the eluting antibody specificfor envelope glycoprotein G2. 2498 CLFNWPVNC(5) CLFTWPASC(1) CLFTWPVSC(1) L-F-[NT]-W-P-[VA]-[NS] 3 Phage display (competitive panning) 5 PMID:9094664 Puumala virus Not determined. Not determined. Not determined. CX7C phage display library Biopanning with MAb 4G2 as the eluting antibody included three antibody and two acid elution rounds. In the first panning, the wells were rinsed 10 times with 5% NFDMP-TBS, pre-eluted with antibody 5A2, rinsed three times, and eluted with MAb 4G2. The second panning was done directly with MAb 4G2, whereas in the third, MAb3H9 was used as a pre-eluting MAb. For removal of residual antibody, the phage amplification product of the third panning was treated with Prosep-A (Bioprocessing Ltd.). The fourth panning was done conventionally with 0.1 M HCl-glycine, pH2.2, containing 1mg of BSA per ml as the eluent. 2499 CPGWWPYPC(1) 1 Phage display (competitive panning) 5 PMID:9094664 Puumala virus Not determined. Not determined. Not determined. CX7C phage display library Biopanning with MAb 4G2 as the eluting antibody included three antibody and two acid elution rounds. In the first panning, the wells were rinsed 10 times with 5% NFDMP-TBS, pre-eluted with antibody 5A2, rinsed three times, and eluted with MAb 4G2. The second panning was done directly with MAb 4G2, whereas in the third, MAb3H9 was used as a pre-eluting MAb. For removal of residual antibody, the phage amplification product of the third panning was treated with Prosep-A (Bioprocessing Ltd.). The fourth and fifth pannings were done conventionally with 0.1 M HCl-glycine, pH2.2, containing 1mg of BSA per ml as the eluent. 2500 CPSWWPVNC(2) CPGWWPYPC(2) CPDWWPYLC(1) P-x-W(2)-P-x(2) 3 Phage display (competitive panning) 3 PMID:9094664 Puumala virus Not determined. Not determined. Not determined. CX7C phage display library The panning included one antibody and two acid elution rounds. All solutions were made with Dul-becco's salt solution containing Ca2+ and Mg2+ (panning, washing, and antibody elution). In the first and second pannings, the phages were allowed to bind to Puumala virus in a solution containing 0.33% BSA. In the first panning, MAb 5B7 was used in the preelution, followed by MAb 1C9 elution, and the well was washed with a 1% BSA solution and then without BSA. Any residual MAb was removed from the amplification product of the first panning with Prosep A. In the second panning, the phages were eluted with a low pH. The third panning was done with Puumala virus-infected cell extract. The extract was diluted 2:3 in phosphate-buffered saline (PBS) containing 0.05% Tween20 (PBST) and 0.5% BSA and captured in a microtiter well coated with MAb 5B7 (5 mg/ml). The phages were added to the well in a solution containing 0.45% BSA and 40 mg of MAb 5A2 per ml, and the well was washed with 0.05% Tween 20. Low-pH elution was then performed. 2501 SSFDQQDWDYSIAEKMHPIRPQFRELPPLPPSRASFGGGASRPSR SSSGYVVPKRLGDMREYNAHPQLHVPPNSPLPPLPTHLQSSRPSR SSRGEGNNIISSRPFLSNSDPWSNKLTGRWGPLPPLPNDSRPSR SSDNWARRVHASELIYTDLSPQILLAQRQLPPTPGRDPSHSRPSR SSYNDLGTRPVSEVIKYDYFPQYSQHVITPDGSYSTRPLPSRPSR SSESPLMYNRVGALQSLTSVQSMMHFALQRRLPRTPPPASRPSR PQYARIVSYRFRALPSPPSASRPSR RPLPPLP 7 Phage display (common panning) PMID:7929027 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. T9 M13 phage display library (X22) 2502 STAPWGLRVAHEGGVLKRPLPIPPVTRPSR STNVWVTGSVIARGAQSRPLPIPPETRPSR STNDVDWMHMWNSGGPHRRLPPTPATRPSR STRWSHSWPGYVGGANPSPATRPLPWPSR STAHSLWDWGTFSGVSHKSRLPPLPTRPSR STAVSFRFMPGGGEAFYSTRPVPPITRPSR STMYGVSWLSSGSGGILAPPVPPRNTRPSR RPLPPLP 7 Phage display (common panning) PMID:7929027 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. T12 M13 phage display library (X36) 2503 LASRPLPLLPNSAPGQ LTGRPLPALPPPFSDF PAYRPLPRLPDLSVIY RALRVRPLPPVPGTSL DAPGSLPFRPLPPVPT LKWRALPPLPETDTPY ISQRALPPLPLMSDPA LTSRPLPDIPVRPSKS NTNRPLPPTPDGLDVR MKDRVLPPIPTVESAV LQSRPLPLPPQSSYPI FINRRLPALPPDNSLL FRALPLPPTPDNPFAG LYSAIAPDPPPRNSSS 14 Phage display (common panning) 3 PMID:8643668 Src SH3 domain of Proto-oncogene tyrosine-protein kinase Src Not determined. Not determined. Not determined. X6PX2PX6 M13 phage display library 2504 ITMRPLPALPGHGQIH LPRRPLPDLPMAAGKG LGSRPLPPTPRQWPEV STIRPLPAIPRDTLLT RSGRPLPPIPEVGHNV IGSRPLPWTPDDLGSA LAQRELPGLPAGAGVS IPGRALPELPPQRALP FVGRELPPTPRTVIPW DPRSALPALPLTPLQT SPHDVLPALPDSHSKS 11 Phage display (common panning) 3 PMID:8643668 SH3 domain of tyrosine-protein kinase Yes Not determined. Not determined. Not determined. X6PX2PX6 M13 phage display library 2505 PPWWAPPPIPNSPQVL PPKFSPPPPPYWQLHA PPHWAPPAPPAMSPPI PPTWTPPKPPGWGVVF PPSFAPPAAPPRHSFG PTYPPPPPPDTAKGA GPRWSPPPVPLPTSLD APTWSPPALPNVAKYK PPDYAAPAIPSSLWVD IKGPRFPVPPVPLNGV PPAWSPPHRPVAFGST APKKPAPPVPMMAHVM 12 Phage display (common panning) 3 PMID:8643668 SH3 domain of tyrosine-protein kinase ABL1 Not determined. Not determined. Not determined. X6PX2PX6 M13 phage display library 2506 LTPQSKPPLPPKPSAV SSHNSRPPLPEKPSWL PVKPPLPAKPWWLPPL TERPPLPQRPDWLSYS LGEFSKPPIPQKPTWM YPQFRPPVPPKPSLMQ VTRPPLPPKPGHMADF VSLGLKPPVPPKPMQL LLGPPVPPKPQTLFSF YKPEVPARPIWLSEL GAGAARPLVPKKPLFL 11 Phage display (common panning) 3 PMID:8643668 SH3 domain of cortactin Not determined. Not determined. Not determined. X6PX2PX6 M13 phage display library 2507 YDASSAPQRPPLPVRKSBR EYVNASPERPPIPGRKSRP WNGIAIPGRPEIPPRASRP SMIFIYPERPSPPPRFSRP GVEEWNPERPQIPLRLSRP WVVDSRPDIPLRRSLP WVPLGRPEIPLRKSLP GGTVGRPPIPERKSVD YSHAGRPEVPPRQSKP FSAAARPDIPSRASTP LYIPKRPEVPPRRHEA NNISARPPLPSRQNPP MAGTPRPAVPQRMNPP 13 Phage display (common panning) 3 PMID:8643668 SH3 domain of apoptosis-stimulating of p53 protein 2 Not determined. Not determined. Not determined. X6PX2PX6 M13 phage display library 2508 MPPPVPPRPPGTLQVA LSYSPPPVPPRPDSTL VLAPPVPPRPGNTFFT YRPPVAPRPPSSLSVD LQCPDCPRVPPRPIPI VPPLVAPRPPSTLNSL LTPPPFPKRPRWTLPE YWPHRPPLAPPQTTLG 8 Phage display (common panning) 3 PMID:8643668 SH3 domains of 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma-1 and gamma-2 Not determined. Not determined. Not determined. X6PX2PX6 M13 phage display library 2509 GQPAGDPDPPPLPAKF FEQTGVPLLPPKSFKY IFGDPPPPIPMKGRSL SNQGSIPVLPIKRVQY NYVNALPPGPPLPAKN SSDPERPVLPPKLWSV HFGPSKPPLPIKTRIT DWKVPEPPVPKLPLKQ ATSEGLPILPSKVGSY NANVSAPRAPAFPVKT EMVLGPPVPPKRGTVV AGSRHPPTLPPKESGG SVAADPPRLPAKSRPQ P-x-L-P-x-K 13 Phage display (common panning) 3 PMID:8643668 SH3 domain of Crk N Not determined. Not determined. Not determined. X6PX2PX6 M13 phage display library 2510 KWDSLLPALPPAFTVE RWDQVLPELPTSKGQI RFDFPLPTHPNLQKAH RLDSPLPALPPTVMQN RWGAPLPPLPEYSWST YWDMPLPRLPGEEPSL RFDYNLPDVPLSLGTA TKKPNAPLPPLPAYMG KWDLDLPPEPMSLGNY YYQRPLPPLPLSHFES YYRKPLPNLPRGQTDD YFDKPLPESPGALMSL YFSRALPGLPERQEAH SLWDPLPPIPQSKTSV SYYDPLPKLPDPGDLG KLYYPLPPVPFKDTKH DPYDALPETPSMKASQ 17 Phage display (common panning) 3 PMID:8643668 SH3 domain of Grb2 N Not determined. Not determined. Not determined. X6PX2PX6 M13 phage display library 2511 SRLDYLKSSLLHLGSR[0.43] SRVDELKLKLSTLVSR[6.1] SRVDYLKDKLISLASR[NT] SRVADVKRKILGLASR[NT] SRVEELRGALLSLKSR[NT] SRLELLKESMRSLASR[NT] SREAWRGRLWDLAKSR[NT] SRVGELKEMMRTLASR[NT] SRLTELKEKLSQLNSR[NT] SRRPEFLKQEIRKASR[NT] 10 Phage display (common panning) 3 PMID:9722581 Troponin C, skeletal muscle Not determined. Not determined. Not determined. X12 M13 phage display ibrary Surface plasmon resonance (SPR) Equilibrium binding constants (μM) for troponin C-binding peptides in the presence of calcium were determined. NT denotes not tested. Nondenaturing polyacrylamide gel electrophoresis demonstrated that pT5 (SRLDYLKSSLLHLGSR) formed a stable complex with troponin C in the presence of calcium. It also showned that the pT5 peptide inhibited the maximal calcium-activated tension of rabbit psoas muscle fibers. 2512 KDYQRNRGPSKFTGLQPSVL 1 Phage display (common panning) 4 PMID:21669956 Purified serum IgG from goat-1 (G1) infection with Map Not determined. Not determined. Not determined. X20 fUSE5 phage display library 2513 VPVRHTPVRDNRLVGFRSSS YVPIVTFYSEISMHSSRAIP EARHVELCPDFKYYPFFCVP 3 Phage display (subtractive panning) 5 PMID:21669956 Purified serum IgG from goat-2 (G2) infection with Map Not determined. Not determined. Not determined. X20 fUSE5 phage display library A pre-clearance step was introduced using serum IgG from goat-2 (G2) prior to (pre-challenge) infection with Map. 2514 VFAEFLPLFSKFGSRMHILK PVLRSGRCAELIQIGFRCRA 2 Phage display (common panning) 4 PMID:16177378 Apical membrane antigen 1, AMA1 Anti-AMA1 monoclonal antibody 4G2dc1 Not determined. Not determined. X20 fUSE5 phage display library 2515 GGWYSFDSPYLMSITEMRLR 1 Phage display (subtractive panning) 3 PMID:16517852 Anti-EBV monoclonal antibody gp125 Epstein-Barr virus, EBV Not determined. Not determined. X20 fUSE5 phage display library Coated wells were blocked with 5% BLOTTO (milk powder diluted in phosphate-buffered saline [PBS]), and the 20-mer phage peptide library was preincubated in 1% BLOTTO for 15 min to remove any milk-binding phage before it was added to the MAb-coated wells. To assess their diagnostic value, a panel of 62 individual EBV IgM sera for their reactivities was screened with the peptides alone. For all peptides, there was a clear distinction between the EBV-positive and the EBV-negative samples, resulting in 100% specificity. The sensitivities were 88%, 85%, 71%, and 54% for peptides F1 (YTDSSMAVTLMKFASNFLF), A3 (SKLLYNYGACRTGCYMAGR, ELISSCLVWSARGCLFGGGI and VMDECVFSSISVLFCNHMLH), gp125 (GGWYSFDSPYLMSITEMRLR), and A2 (DNYWSFSDSTYWTLRYSSG), respectively. Any combination of peptides increased the sensitivity, indicating that individual peptides react with different subsets of antibodies. Furthermore, when the F1 and the gp125 peptides were coupled to bovine serum albumin and screened against 216 serum samples, there were dramatic improvements in sensitivities (95% and 92%, respectively) and little cross-reactivity with the other peptides encountered during acute viral infections, including rheumatoid factor. This study shows the potential for the use of peptide mimotopes as alternatives to the complex antigens used in current serodiagnostics for EBV infection. 2516 YTDSSMAVTLMKFASNFLF 1 Phage display (subtractive panning) 3 PMID:16517852 Anti-EBV monoclonal antibody F1 Epstein-Barr virus, EBV Not determined. Not determined. X20 fUSE5 phage display library Coated wells were blocked with 5% BLOTTO (milk powder diluted in phosphate-buffered saline [PBS]), and the 20-mer phage peptide library was preincubated in 1% BLOTTO for 15 min to remove any milk-binding phage before it was added to the MAb-coated wells. To assess their diagnostic value, a panel of 62 individual EBV IgM sera for their reactivities was screened with the peptides alone. For all peptides, there was a clear distinction between the EBV-positive and the EBV-negative samples, resulting in 100% specificity. The sensitivities were 88%, 85%, 71%, and 54% for peptides F1 (YTDSSMAVTLMKFASNFLF), A3 (SKLLYNYGACRTGCYMAGR, ELISSCLVWSARGCLFGGGI and VMDECVFSSISVLFCNHMLH), gp125 (GGWYSFDSPYLMSITEMRLR), and A2 (DNYWSFSDSTYWTLRYSSG), respectively. Any combination of peptides increased the sensitivity, indicating that individual peptides react with different subsets of antibodies. Furthermore, when the F1 and the gp125 peptides were coupled to bovine serum albumin and screened against 216 serum samples, there were dramatic improvements in sensitivities (95% and 92%, respectively) and little cross-reactivity with the other peptides encountered during acute viral infections, including rheumatoid factor. This study shows the potential for the use of peptide mimotopes as alternatives to the complex antigens used in current serodiagnostics for EBV infection. 2517 DNYWSFSDSTYWTLRYSSG 1 Phage display (subtractive panning) 3 PMID:16517852 Anti-EBV monoclonal antibody A2 Epstein-Barr virus, EBV Not determined. Not determined. X20 fUSE5 phage display library Coated wells were blocked with 5% BLOTTO (milk powder diluted in phosphate-buffered saline [PBS]), and the 20-mer phage peptide library was preincubated in 1% BLOTTO for 15 min to remove any milk-binding phage before it was added to the MAb-coated wells. To assess their diagnostic value, a panel of 62 individual EBV IgM sera for their reactivities was screened with the peptides alone. For all peptides, there was a clear distinction between the EBV-positive and the EBV-negative samples, resulting in 100% specificity. The sensitivities were 88%, 85%, 71%, and 54% for peptides F1 (YTDSSMAVTLMKFASNFLF), A3 (SKLLYNYGACRTGCYMAGR, ELISSCLVWSARGCLFGGGI and VMDECVFSSISVLFCNHMLH), gp125 (GGWYSFDSPYLMSITEMRLR), and A2 (DNYWSFSDSTYWTLRYSSG), respectively. Any combination of peptides increased the sensitivity, indicating that individual peptides react with different subsets of antibodies. Furthermore, when the F1 and the gp125 peptides were coupled to bovine serum albumin and screened against 216 serum samples, there were dramatic improvements in sensitivities (95% and 92%, respectively) and little cross-reactivity with the other peptides encountered during acute viral infections, including rheumatoid factor. This study shows the potential for the use of peptide mimotopes as alternatives to the complex antigens used in current serodiagnostics for EBV infection. 2518 SKLLYNYGACRTGCYMAGR ELISSCLVWSARGCLFGGGI VMDECVFSSISVLFCNHMLH 3 Phage display (subtractive panning) 3 PMID:16517852 Anti-EBV monoclonal antibody A3 Epstein-Barr virus, EBV Not determined. Not determined. X20 fUSE5 phage display library Coated wells were blocked with 5% BLOTTO (milk powder diluted in phosphate-buffered saline [PBS]), and the 20-mer phage peptide library was preincubated in 1% BLOTTO for 15 min to remove any milk-binding phage before it was added to the MAb-coated wells. To assess their diagnostic value, a panel of 62 individual EBV IgM sera for their reactivities was screened with the peptides alone. For all peptides, there was a clear distinction between the EBV-positive and the EBV-negative samples, resulting in 100% specificity. The sensitivities were 88%, 85%, 71%, and 54% for peptides F1 (YTDSSMAVTLMKFASNFLF), A3 (SKLLYNYGACRTGCYMAGR, ELISSCLVWSARGCLFGGGI and VMDECVFSSISVLFCNHMLH), gp125 (GGWYSFDSPYLMSITEMRLR), and A2 (DNYWSFSDSTYWTLRYSSG), respectively. Any combination of peptides increased the sensitivity, indicating that individual peptides react with different subsets of antibodies. Furthermore, when the F1 and the gp125 peptides were coupled to bovine serum albumin and screened against 216 serum samples, there were dramatic improvements in sensitivities (95% and 92%, respectively) and little cross-reactivity with the other peptides encountered during acute viral infections, including rheumatoid factor. This study shows the potential for the use of peptide mimotopes as alternatives to the complex antigens used in current serodiagnostics for EBV infection. 2519 GGWYSFDSPYLMSITEMRLR 1 Phage display (subtractive panning) 4 PMID:16517852 Anti-EBV monoclonal antibody gp125 Epstein-Barr virus, EBV Not determined. Not determined. X20 fUSE5 phage display library Coated wells were blocked with 5% BLOTTO (milk powder diluted in phosphate-buffered saline [PBS]), and the 20-mer phage peptide library was preincubated in 1% BLOTTO for 15 min to remove any milk-binding phage before it was added to the MAb-coated wells. To assess their diagnostic value, a panel of 62 individual EBV IgM sera for their reactivities was screened with the peptides alone. For all peptides, there was a clear distinction between the EBV-positive and the EBV-negative samples, resulting in 100% specificity. The sensitivities were 88%, 85%, 71%, and 54% for peptides F1 (YTDSSMAVTLMKFASNFLF), A3 (SKLLYNYGACRTGCYMAGR, ELISSCLVWSARGCLFGGGI and VMDECVFSSISVLFCNHMLH), gp125 (GGWYSFDSPYLMSITEMRLR), and A2 (DNYWSFSDSTYWTLRYSSG), respectively. Any combination of peptides increased the sensitivity, indicating that individual peptides react with different subsets of antibodies. Furthermore, when the F1 and the gp125 peptides were coupled to bovine serum albumin and screened against 216 serum samples, there were dramatic improvements in sensitivities (95% and 92%, respectively) and little cross-reactivity with the other peptides encountered during acute viral infections, including rheumatoid factor. This study shows the potential for the use of peptide mimotopes as alternatives to the complex antigens used in current serodiagnostics for EBV infection. 2520 YTDSSMAVTLMKFASNFLF 1 Phage display (subtractive panning) 4 PMID:16517852 Anti-EBV monoclonal antibody F1 Epstein-Barr virus, EBV Not determined. Not determined. X20 fUSE5 phage display library Coated wells were blocked with 5% BLOTTO (milk powder diluted in phosphate-buffered saline [PBS]), and the 20-mer phage peptide library was preincubated in 1% BLOTTO for 15 min to remove any milk-binding phage before it was added to the MAb-coated wells. To assess their diagnostic value, a panel of 62 individual EBV IgM sera for their reactivities was screened with the peptides alone. For all peptides, there was a clear distinction between the EBV-positive and the EBV-negative samples, resulting in 100% specificity. The sensitivities were 88%, 85%, 71%, and 54% for peptides F1 (YTDSSMAVTLMKFASNFLF), A3 (SKLLYNYGACRTGCYMAGR, ELISSCLVWSARGCLFGGGI and VMDECVFSSISVLFCNHMLH), gp125 (GGWYSFDSPYLMSITEMRLR), and A2 (DNYWSFSDSTYWTLRYSSG), respectively. Any combination of peptides increased the sensitivity, indicating that individual peptides react with different subsets of antibodies. Furthermore, when the F1 and the gp125 peptides were coupled to bovine serum albumin and screened against 216 serum samples, there were dramatic improvements in sensitivities (95% and 92%, respectively) and little cross-reactivity with the other peptides encountered during acute viral infections, including rheumatoid factor. This study shows the potential for the use of peptide mimotopes as alternatives to the complex antigens used in current serodiagnostics for EBV infection. 2521 DNYWSFSDSTYWTLRYSSG 1 Phage display (subtractive panning) 4 PMID:16517852 Anti-EBV monoclonal antibody A2 Epstein-Barr virus, EBV Not determined. Not determined. X20 fUSE5 phage display library Coated wells were blocked with 5% BLOTTO (milk powder diluted in phosphate-buffered saline [PBS]), and the 20-mer phage peptide library was preincubated in 1% BLOTTO for 15 min to remove any milk-binding phage before it was added to the MAb-coated wells. To assess their diagnostic value, a panel of 62 individual EBV IgM sera for their reactivities was screened with the peptides alone. For all peptides, there was a clear distinction between the EBV-positive and the EBV-negative samples, resulting in 100% specificity. The sensitivities were 88%, 85%, 71%, and 54% for peptides F1 (YTDSSMAVTLMKFASNFLF), A3 (SKLLYNYGACRTGCYMAGR, ELISSCLVWSARGCLFGGGI and VMDECVFSSISVLFCNHMLH), gp125 (GGWYSFDSPYLMSITEMRLR), and A2 (DNYWSFSDSTYWTLRYSSG), respectively. Any combination of peptides increased the sensitivity, indicating that individual peptides react with different subsets of antibodies. Furthermore, when the F1 and the gp125 peptides were coupled to bovine serum albumin and screened against 216 serum samples, there were dramatic improvements in sensitivities (95% and 92%, respectively) and little cross-reactivity with the other peptides encountered during acute viral infections, including rheumatoid factor. This study shows the potential for the use of peptide mimotopes as alternatives to the complex antigens used in current serodiagnostics for EBV infection. 2522 SKLLYNYGACRTGCYMAGR ELISSCLVWSARGCLFGGGI VMDECVFSSISVLFCNHMLH 3 Phage display (subtractive panning) 4 PMID:16517852 Anti-EBV monoclonal antibody A3 Epstein-Barr virus, EBV Not determined. Not determined. X20 fUSE5 phage display library Coated wells were blocked with 5% BLOTTO (milk powder diluted in phosphate-buffered saline [PBS]), and the 20-mer phage peptide library was preincubated in 1% BLOTTO for 15 min to remove any milk-binding phage before it was added to the MAb-coated wells. To assess their diagnostic value, a panel of 62 individual EBV IgM sera for their reactivities was screened with the peptides alone. For all peptides, there was a clear distinction between the EBV-positive and the EBV-negative samples, resulting in 100% specificity. The sensitivities were 88%, 85%, 71%, and 54% for peptides F1 (YTDSSMAVTLMKFASNFLF), A3 (SKLLYNYGACRTGCYMAGR, ELISSCLVWSARGCLFGGGI and VMDECVFSSISVLFCNHMLH), gp125 (GGWYSFDSPYLMSITEMRLR), and A2 (DNYWSFSDSTYWTLRYSSG), respectively. Any combination of peptides increased the sensitivity, indicating that individual peptides react with different subsets of antibodies. Furthermore, when the F1 and the gp125 peptides were coupled to bovine serum albumin and screened against 216 serum samples, there were dramatic improvements in sensitivities (95% and 92%, respectively) and little cross-reactivity with the other peptides encountered during acute viral infections, including rheumatoid factor. This study shows the potential for the use of peptide mimotopes as alternatives to the complex antigens used in current serodiagnostics for EBV infection. 2523 DGPSYHVAFKNSRGLRHS 1 Phage display (subtractive panning) 4-6 PMID:19073711 Purified serum IgG from an EBV-infected patient Epstein-Barr virus, EBV Not determined. Not determined. X20 fUSE5 phage display library The 20-mer phage peptide library was preincubated in BLOTTO to remove any milk-binding phage before it was added to the purified human anti-EBV IgG preparations-coated wells. 2524 NGALYPRFFPDYSILMFPII DQFAQAYRGDRNFFNELTST RQFSKFKDASDRYGNYLHFF SSSIKIWNKLGWNTVIAGTR FVNAFQNANFMRPRELFALA SANLNFFSPDFGLYTPNASA AITCAHTLSIKSRRCQYVFK AASYASRTVGFASVYWFSRP RLRGDYNVGPIRFGWPVAPN MSDFDRKVYTFNFITDPQHL GVTDFDFKVFSSTFPKIFLS TPNTVRDFYYNVSLPSYMLI 12 Phage display (subtractive panning) 4-6 PMID:19073711 Purified serum IgG from an EBV-immunised rabbit Epstein-Barr virus, EBV Not determined. Not determined. X20 fUSE5 phage display library The 20-mer phage peptide library was preincubated in BLOTTO to remove any milk-binding phage before it was added to the purified rabbit anti-EBV IgG preparations-coated wells. 2525 LPPNPTK(2) DPGATPP(1) TLTPPPL(1) PQSSPPH(1) TNRLHPP(1) TRLPPLS(1) LPLSGND(1) APPGRPT(1) AKALPHT(1) APRHNTQ(1) AAPLHNK(1) [TLAH]-P-P 11 Phage display (subtractive panning) 3 http://d.g.wanfangdata.com.cn/Periodical_zglx-e200504001.aspx Endoglucanase isozyme AgEGl, IsoA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The phage display library was incubated first with native polyacrylamide gel slices to remove phage clones binding to polyacrylamide gel. 2526 IWAADWV IWQWEWL VWSVEWV PWMGPEW 4 Phage display (common panning) 3 PMID:17113859 Anti-Lewis X monoclonal antibody MMA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2527 LWKFMGR IWRWAYF 2 Phage display (common panning) 3 PMID:17113859 Anti-Lewis X monoclonal antibody SSEA-1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2528 IWLQRRH LYLQRLR MWLQSVH YWLQTVR LWLMKAR 5 Phage display (common panning) 3 PMID:17113859 Anti-Lewis X monoclonal antibody Pmn6 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2529 VHHSCTKLTHCCQNWH(11/25) GGVSCMQTSPVCENNL(6/25) TKRDCSAQNYGCQKAI(2/25) 3 Phage display (subtractive panning) 2 PMID:18723274 Tumor-associated glycoprotein (TAG-72) Not determined. Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) To begin one selection cycle, the three coated wells of one plate were washed and then blocked with 0.1 M sodium bicarbonate buffer containing 0.5% BSA. The phage library, containing about 2e8 phages in 400 μl of TBS/Tween and 0.1% BSA, was added first to the well coated with the TAG-72 free eluant from the affinity column. After incubation for 20 min the library was transferred and incubated for 30 min in the BSA coated well. Thereafter, the library was transferred to the final well containing the purified TAG-72 and incubated with gentle mixing for 1 hr at room temperature. 2530 VHHSCTKLTHCCQNWH(0.52) GGVSCMQTSPVCENNL(0.24) 2 Phage display (subtractive panning) 3 PMID:18723274 Tumor-associated glycoprotein (TAG-72) Not determined. Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) To begin one selection cycle, the three coated wells of one plate were washed and then blocked with 0.1 M sodium bicarbonate buffer containing 0.5% BSA. The phage library, containing about 2e8 phages in 400 μl of TBS/Tween and 0.1% BSA, was added first to the well coated with the TAG-72 free eluant from the affinity column. After incubation for 20 min the library was transferred and incubated for 30 min in the BSA coated well. Thereafter, the library was transferred to the final well containing the purified TAG-72 and incubated with gentle mixing for 1 hr at room temperature. 2531 RNVPPTFNDVYWIAF VPPCFTLMYCAGVVR V-P(2)-x-F-x(3)-Y 2 Phage display (common panning) 3 PMID:9414130 Anti-Lc4Cer monoclonal antibody AD117m Not determined. Not determined. Not determined. X15 fUSE5 phage display library The antibody was biotinylated by incubation with sulfo-NHS-biotin (Pierce, Rockford, IL). Each round of biopanning was performed on the streptavidin-coated polystyrene Petri dish. 2532 FRSFHYHTGRWHWLR RNVPPIFNDVYWIAF ARFPKELRGSVRSAH 3 Phage display (common panning) 3 PMID:9414130 Anti-nLc4Cer monoclonal antibody H11 Not determined. Not determined. Not determined. X15 fUSE5 phage display library The antibody was biotinylated by incubation with sulfo-NHS-biotin (Pierce, Rockford, IL). Each round of biopanning was performed on the streptavidin-coated polystyrene Petri dish. 2533 FRSDVRFWHWSTPFM(11) VRVYFGFGPPPYFGG(9) WHWRHRIPLQLAAGR(2) RYWLYGDPASFPVNH(2) 4 Phage display (common panning) 4 PMID:9877157 Anti-GD1α monoclonal antibody KA17 Ganglioside GD1α SIAT7-E and SIAT7-F Not determined. Not determined. X15 fUSE5 phage display library Each round of biopanning was performed on the 96-well microtiter well (Falcon 3072) coated with streptavidin. The biotinylated KA17 was adsorbed onto the streptavidin-coated well, and affinity-isolation of phages from the peptide epitope library by four cycles of biopanning was carried out. 2534 ISLLQAR IDLMQAR IILLQAR IELLQAR ISLLGAR FSLLDAR IFLLWQR FAQLDWH 8 Phage display (common panning) 3 PMID:10667600 Anti-Lewis A monoclonal antibody Sialyl Lewis A oligosaccharide Not determined. Not determined. f3-7mer fUSE5-based phage display library A flat-bottomed Linbro/Titertek 96-well plate (ICN Biomedicals) was coated with 10 mg of goat antimouse immunoglobulins, blocked with PBS containing 3% BSA and 0.02% Tween 20, and then incubated with 2-3 mg monoclonal anti-Lewis A antibody (clone 7LE; mouse IgG1, Seikagaku Biochemicals). After washing with PBS containing 0.02% Tween 20, wells were blocked with PBS containing 3% BSA. A linear heptapeptide phage library (1e1-1e12 transducing units) was added to the well and incubated at room temperature for 1 h. After removing unbound phage, phages eluted in the first round were amplified and subjected to another two rounds of paning with anti-Lewis A antibody in the same manner as the first selection. 2535 DRPVPY[765] KSPTPY[641] ARPLWY[583] VRPQVP[524] R-P-x(2)-Y 4 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody SA3 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. X6 phage display library ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector fd-tet without a random peptide insert is 169. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2536 MCPPLYSPSACA[957] ECNFLYPGFTCA[202] C-x(1-2)-L-Y 2 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody Strep 9 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. LX-8 phage display library (XCX8CX) ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 14. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2537 YPYCGHALCPGLYADAS[1020] VILPYDNNCALCLNLYP[644] VIDAPTPNCAWPNGRRG[256] MPPAGTGTCFLYALSCS[153] ADLSPTPYCQPSTMHTN[144] NEYINQDHCLLYAMLCP[38] P-T-P-x-C 6 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody Strep 9 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. X8CX8 f88-based phage display library ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 14. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2538 EIAPQGPSKCLLYAYCQ[13] 1 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody Strep 9 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. X15CX phage display library ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 14. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2539 ADAAPSPTPYLPRLS[643] ATYRPVPAEFARKHL[425] TITATDSPTPWPFER[244] P-x-P-x(1-2)-P 3 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody HGAC 39 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. X15 phage display library ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 72. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2540 MCRPSPYNPPCT[112] 1 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody HGAC 39 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. LX-8 phage display library (XCX8CX) ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 72. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2541 MCRPGIPTHHCA[174] HCSPGQRPGTCQ[165] DCGNMLHAEVCR[149] DCRPGVPLLSCP[115] RPG 4 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody HGAC 47 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. LX-8 phage display library (XCX8CX) ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 87. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2542 SCISAACFCI[141] 1 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody HGAC 101 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. f88-LX6 phage display library (XCX6CX) ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 29. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2543 KQLMAP[138] 1 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody HGAC 101 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. X6 phage display library ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 29. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2544 NYPMDH[141] MYPMSH[104] YPMGHL[27] IYPMPA[13] QQYPMG[12] QSTYPM[10] YPM 6 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody SE155.4 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. X6 phage display library ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 14. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2545 EPYPMSEANYVRPMP[267] YPMPASSDNAQWLLK[15] DGTNAYPMNEDISVS[15] HSTRNYSYLGSPYPM[13] NYPMSGARIEPLLHA[13] YAATEPRYMIPYPMP[13] YPMGETCQRIRSCVW[11] YPM 7 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody SE155.4 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. X15 phage display library ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 14. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2546 VCPAPYPAGTCA[11] 1 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody SE155.4 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. LX-8 phage display library (XCX8CX) ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 14. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2547 YTTQCGYGGCMNFE[938] MGVICMNMECDRNM[799] LHEYCNMETCPYNH[585] QYPQCHNMDCKSIT[472] PTHVCYNMECQGGD[275] TPTNCYNMTCQNQP[187] C-x-N-M-[ED] 6 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody SYAJ/6 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. X4CX4CX4 f88-4-based phage display library ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 66. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2548 MDWNMH[993] 1 Phage display (common panning) 3-4 PMID:9122216 Anti-GAS CWPS monoclonal antibody SYAJ/6 Cell-wall polysaccharide (CWPS) of group A Streptococcus (GAS), GAS CWPS Not determined. Not determined. X6 phage display library ELISA Absorbances are calculated as (A405 - A490) × 1000. The value of the wild-type phage vector f88-4 without a random peptide insert is 66. The library was affinity-selected on biotinylated mAbs that had been immobilized in avidin- or streptavidin-coated microwells. 2549 KSQLPWT[25.0] KSADLKR[0.5] KSCVRGR[6.5] KSCDRGR[NT] KSYCRTR[NT] KSNSQHP[NT] KSCQMDS[NT] QSQVTRT[NT] TSQSQSP[NT] VSHIQAN[NT] 10 Phage display (common panning) 5 PMID:7831288 Hairpin DNA containing an altered Zif268 binding site GCG Not determined. Not determined. Not determined. pComb3.5 XSX5 phage display library Surface plasmon resonance (SPR) The calculated equilibrium dissociation constants (Kd, nM) of zinc-finger proteins are shown. The value of the wild-type (WT) protein is 6.5. NT denotes not tested. 2550 QSTASKA[29.7] PSTHLQT[41.6] PSERTQP[NT] TSSEADH[NT] SSEQRYP[NT] HSQQNKP[NT] RSGQGMA[NT] RSARQTG[NT] ESNSFTD[NT] NSVMGHD[NT] NSRGQRK[NT] SSRPSQW[NT] TSSEADH[NT] 13 Phage display (common panning) 5 PMID:7831288 Hairpin DNA containing an altered Zif268 binding site TGT Not determined. Not determined. Not determined. pComb3.5 XSX5 phage display library Surface plasmon resonance (SPR) The calculated equilibrium dissociation constants (Kd, nM) of zinc-finger proteins are shown. The value of the wild-type (WT) protein is 81.8. NT denotes not tested. 2551 TYLNTP GYRAAP LYRYHL PTLVNA VRPHQR PFCPYR 6 Phage display (common panning) 5 PMID:7831288 Hairpin DNA containing an altered Zif268 binding site TGG Not determined. Not determined. Not determined. pComb3.5 X6 phage display library 2552 GVTMQP[15.9] REQVSR[6.4] QRMRTL[27.5] LRTGNY[4.6] PQPLSD[NT] THMWMI[NT] QRVGLF[NT] EREFSL[NT] EKESRG[NT] EGVRKN[NT] TGVNSI[NT] TQARPP[NT] THMWMI[NT] 13 Phage display (common panning) 5 PMID:7831288 Hairpin DNA containing an altered Zif268 binding site TTG Not determined. Not determined. Not determined. pComb3.5 X6 phage display library Surface plasmon resonance (SPR) The calculated equilibrium dissociation constants (Kd, nM) of zinc-finger proteins are shown. The value of the wild-type (WT) protein is 83.7. NT denotes not tested. 2553 SSGQWWR[13.1] RSDLANS[NT] SSLLVIA[NT] VSSVRGL[NT] 4 Phage display (common panning) 5 PMID:7831288 Hairpin DNA containing an altered Zif268 binding site GCG Not determined. Not determined. Not determined. pComb3.5 XSX5 phage display library Surface plasmon resonance (SPR) The calculated equilibrium dissociation constants (Kd, nM) of zinc-finger proteins are shown. The value of the wild-type (WT) protein is 6.5. NT denotes not tested. 2554 NSVGDKP SSWICGI ISAWMEL ISMMTFF RSECRML ISALLDT NSVQGLR 7 Phage display (common panning) 5 PMID:7831288 Hairpin DNA containing an altered Zif268 binding site CTG Not determined. Not determined. Not determined. pComb3.5 XSX5 phage display library 2555 SRSDHLIR(2) SRSDHLTR(1) SRSDHLLR(1) SRTDHLTR(1) SRSDHLTT(1) 5 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GGG. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2556 SRSDNLRR(5) 1 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GAG. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2557 ARRDNLQR(1) 1 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H X5LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GAG. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2558 GRHDSLLR(5) ARKDALTR(1) 2 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H X5LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GTG. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2559 SRKDSLVR(1) SRMDSLAR(1) SRLDSLKR(1) SRGDALRR(1) SRSDSLLR(1) 5 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GTG. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2560 SRLDTLGR(5) SRRDTLVR(1) SRKDTLAR(1) SRSDTLRR(1) SRSDTLKR(1) 5 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GCG. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2561 GRADTLRR(3) 1 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H X5LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GCG. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2562 SQRAHLER(3) SQAGHLRR(3) 2 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GGA. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2563 SQRSNLVR(6) 1 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GAA. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2564 SQSSSLVR(2) SERSSLVR(2) SQLSGLKR(1) SQMGSLTR(1) SQRGSLLR(1) 5 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GTA. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2565 SQSGDLRR(4) SQSGDLQR(1) SQKGTLIR(1) SQQGTLRR(1) SRSDHLTT(1) 5 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GCA. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2566 AQSGDLTR(1) AQAGTLMR(1) 2 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H X5LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GCA. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2567 TTADKLSR(3) RTKSSLRR(2) TTTGGLTR(1) 3 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H X5LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GGT. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2568 STQGGLAR(1) STSGSLAR(1) STTGGLAR(1) STSGGLAR(1) STSGGLTR(1) 5 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GGT. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2569 STSGNLVR(2) STEGNLVR(2) STSGNLKR(1) STKGNLAR(1) 4 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GAT. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2570 STSGSLTR(6) STSGALTT(4) SERWMLLR(2) 3 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GTT. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2571 ARRSDLTR(4) 1 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H X5LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GCT. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2572 SSSQTLTR(2) 1 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GCT. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2573 SERSKLAR(6) 1 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GGC. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2574 RDPGNLKR(5) 1 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H X5LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GAC. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2575 SAKRMLGQ(1) 1 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GAC. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2576 SDPGALVR(5) 1 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GTC. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2577 TEKGTLNR(1) 1 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H X5LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GTC. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2578 SGCRELSR(5) SDCRDLAR(4) SGCKDLAR(1) SGCRELSR(1) 4 Phage display (subtractive panning) 7 PMID:10077584 Hairpin DNA Not determined. Not determined. Not determined. pComb3H SX4LX2 phage display library Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GCC. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads. 2579 EERYMNVMPFGGGGS(5) EEQYVNMPWFGGGGS(4) EDERYMNLPWGGGGS(3) QLYENWPVLTGGGGS(2) QERYENVPGIGGGGS(1) RERYENVWYVGGGGS(1) RSGYENWPVIGGGDS(1) GDEHYRNSLGGGGS(1) SSERYENVIFGGGGS(1) QREKYENWPFGGGGS(1) Y-x-N 10 Phage display (subtractive panning) 4 PMID:10400318 The SH2 domain of growth factor receptor-bound protein 2 Not determined. Not determined. Not determined. X10 phagemid-based fd phage display library Both MBP and GST fusions were used as affinity selection reagents in the experiments. To enhance the possibility of isolating SH2 domain-specific phage, the fusion partner-specific phage were "subtracyed" by co-incubating the phage with soluble GST or MBP during the affinity selection. 2580 ECYINVPFTCMAGASAG(7) TECYKNVPEICAGASAG(1) Y-x-N 2 Phage display (subtractive panning) 4 PMID:10400318 The SH2 domain of growth factor receptor-bound protein 2 Not determined. Not determined. Not determined. X12 phagemid-based fd phage display library Both MBP and GST fusions were used as affinity selection reagents in the experiments. To enhance the possibility of isolating SH2 domain-specific phage, the fusion partner-specific phage were "subtracyed" by co-incubating the phage with soluble GST or MBP during the affinity selection. 2581 GGCDEVYVNWSCGG(8) GGCPERYENVMCGG(4) GGCLSYMNSPMCEG(2) GGCYENLWPYSCGG(2) GGYDELYENWPCGG(2) GGCDGYYENWDCGG(1) GGCMEEYVNWSCGG(1) GGCQEEYVNWSCGG(1) GGCVHYENYMWCGG(1) GGCHYVNWPPECGG(1) GGCYVNVYDPLCGG(1) GGCVLYENGTFCGG(1) GGCYWQNVPESCGG(1) Y-x-N 13 Phage display (subtractive panning) 4 PMID:10400318 The SH2 domain of growth factor receptor-bound protein 2 Not determined. Not determined. Not determined. CX8C phagemid-based fd phage display library Both MBP and GST fusions were used as affinity selection reagents in the experiments. To enhance the possibility of isolating SH2 domain-specific phage, the fusion partner-specific phage were "subtracyed" by co-incubating the phage with soluble GST or MBP during the affinity selection. 2582 RSCGLDYEGYWLKLK(13) GGGQWLGTWEWYGPK(10) YEKISVEGIKDVRVR(9) NVSIEGVLKYYRGLR(6) SRWLEEEVSRLLLLR(6) GEALDRFEREMKLMR(1) 6 Phage display (common panning) 7 PMID:10449733 DNA duplex containing an inverted repeat of the Zif12-binding site Not determined. Not determined. Not determined. pZif12 X15 phage display library 2583 FSRPKRPPHW(3) FKPRRPPPPG(3) FPRRPPRPRL(1) P-[RK]-R-P(2)-x-P 3 Phage display (common panning) 3 PMID:10542231 SH3 domain of endophilin-A1 Not determined. Not determined. Not determined. X9 PC89-based phage display library GST-SH3 fusion protein was the real target. 2584 FPKRPPLPRS(8) FSRPKRPPHW(2) FPRRPPPPRM(1) P-[RK]-R-P(2)-x-P-R 3 Phage display (common panning) 3 PMID:10542231 SH3 domain of endophilin-A2 Not determined. Not determined. Not determined. X9 PC89-based phage display library GST-SH3 fusion protein was the real target. 2585 FSRPKRPPHW(3) FPRRPPRPRL(3) FPPRPPGPYA(1) FQKPSRPKPW(1) FPPRPPAPYW(1) FQKPQRPPRW(1) FPPRPKPPRM(1) FPKRPPLPRS(1) FRPPRPPLP(1) P-x-R-P(2)-x-P-R 9 Phage display (common panning) 3 PMID:10542231 SH3 domain of endophilin-A3 Not determined. Not determined. Not determined. X9 PC89-based phage display library GST-SH3 fusion protein was the real target. 2586 FVRPARRVLW(1) FHRPVRRAAP(1) FVRPTRAADA(1) FXRPRRGHAL(1) FHRPTRSKLA(1) FKRPPRDGTL(1) FQRPLRRPRM(1) FFPKRPPRIL(1) FRRPRRSLPE(1) FTRPYRPTHP(1) FGRPPRNARV(1) FHRPWRRWRE(1) x(2)-R-P-x-R 12 Phage display (common panning) 3 PMID:10542231 SH3 domain of amphiphysin Not determined. Not determined. Not determined. X9 PC89-based phage display library GST-SH3 fusion protein was the real target. 2587 SSVICSEGPGGWSCVRWE GVWRCILGPDGWLCLAVQ GYEWRCWPTRGQWLCILTGL GAKLVCATDWVVILCHKVVT WDTLLCKHYDVRWICSLITR ATESQCYWSESGVLCWVTGP SSMYWCATLETMWWCWRVVE SKGWLCIWRVPGYVCIKFWT MEELHCARDGSQLWCWWGVL EWYQLCATGPRGSRCWWVQL HREFLCWSLGEEGARCFVIW EVMRTCYRAWEWGWICLLQA IEWECIALRDHVWRCWVPLP 13 Phage display (common panning) 6 PMID:12618188 Elongation factor Tu, EF-Tu Not determined. Not determined. Not determined. XnCXnCXn and Xn M13 phage display library pool In the fourth round, the amplified phages were either injected to the biosensor (BIAcore 2000). In the biosensor, the phages flowed first through a sensor chip to which MAb 5A2 was coupled and then through a control sensor chip to which MAb 5B7 was coupled (about 5 ng of each antibody). 2588 CWDDGWLC(4) YLCHARAIDTTCIWPEPCFD(3) WGCVLHMVDDGRLGAVICGS(3) RMCLQCRIGSGCRAVLCAY(2) PMCSNRMADDIWSVPWYCVR(1) RLCSVSYPEPSLLWTSSCFD(1) RLCAGGASLIEPVYALRCAE(1) VACSYPEPYLSVAAGCNQ(1) EQCWKLFPANCVGCATRCNA(1) DLCHLRKTEGIWAMHCV(1) 10 Phage display (competitive panning) 4 PMID:10502511 Anti-Puumala virus envelope glycoprotein G1 monoclonal antibody 5A2 Not determined. Not determined. Not determined. X2CX14CX2 phage display library 2589 RMCLQCRIGSGCRAVLCAY(1) 1 Phage display (common panning) 4 PMID:10502511 Anti-Puumala virus envelope glycoprotein G1 monoclonal antibody 5A2 Not determined. Not determined. Not determined. X2CX14CX2 phage display library 2590 RMCLQCRIGSGCRAVLCAY(5) EQCWKLFPANCVGCATRCNA(1) 2 Phage display (common panning) 4 PMID:10502511 Anti-Puumala virus envelope glycoprotein G1 monoclonal antibody 5A2 Not determined. Not determined. Not determined. X2CX14CX2 phage display library 2591 SLCAGQVVFERTAARCSQ(2) RGCIKKSDRPEKRLASGCGV(1) INCRIGTRLLRDAPAQVCWA(1) ASCPVQQYPSGPCEASVCDT(1) GPCVEERHWLGRRDSILCNR(1) LMCGWTMDPVWGAWTLTCTA(1) LRCLGGHNESASRRAIECTP(1) ERCKMMVPGMTGSAALVCPM(1) GHCALWTSPHFRIEARLCGL(1) 9 Phage display (competitive panning) 2 PMID:10502511 Anti-Puumala virus envelope glycoprotein G2 monoclonal antibody 4G2 Not determined. Not determined. Not determined. X2CX14CX2 phage display library In the first round, the library was allowed to react with 60 ng of MAb 4G2 coated on a microtiter well. The microtiter well was extensively washed with virus antigen. The phages were finally eluted competitively with the virus antigen. In the second round, the panning reaction was done in 1% BSA PBST in the presence of 3 mg/ml the bank vole anti-PUU N MAb 5E1. 2592 FPCDRLSGYWERGIPSPCVR(9) LYCLVYVPEVQTMYCDG(1) CYCTRLHEDGSYAPTFYCLL(1) LYCISSGGKMKDASTVYCRN(1) 4 Phage display (competitive panning) 3 PMID:10502511 Anti-Puumala virus envelope glycoprotein G2 monoclonal antibody 1C9 Not determined. Not determined. Not determined. X2CX14CX2 phage display library Negative serum in the liquid phase was used to compete the binding of phages to 1C9 in the two first panning rounds. 2593 SRAGLLSDLLEGKSR SSRSLLRDLLMVDSR SSNKLLYNLLKMESR SSKSLLLNLLSTPSR SRLEMLLRSETDFSR SRLEELLKWGSVTSR SRLEQLLKEEFSYSR SRLEQLLRSEPDFSR SRLEDLLRAPFTTSR SRLESLLRFGQLDSR SSRLLSLLVGDFNSR SRLEELLLGTNRDSR SRLEELLLMDFWRSR SRLKELLLLPTDLSR SRLECLLEGRLNCSR SSKLYCLLDESYCSR SRLSCLLMGFEDCSR SSNHQSSRLIELLSR SSRLWQLLASTDTSR SSKLWQLLSSPIDSR SRLVALLKSPWSVSR SSNSMLWKLLAAPSR SSKTLWRLLEGERSR SRAGPVLWGLLSESR SRSPILTHLLSLGSR SSTGILWKLLTAESR SSHGILWRLLSEGSR SSKLIRLLTSDEELSR SSRLMELLQEGQGWSR HSFPRESLLVRLLQGG L-x(2)-L(2) 30 Phage display (common panning) PMID:10097152 Estrogen receptor alpha, ERα Not determined. Not determined. Not determined. Not determined. Immulon 4 96-well plates were coated with streptavidin in 0.1 M sodium bicarbonate. The plates were then incubated for 1 h with 2 pmol biotinylated vitellogenin ERE per well, followed by incubation for 1 h with 3 pmol (monomer) ERα per well. Affinity selections were conducted with the ER in TBST (10 mM Tris.HCl, pH 8.0/150 mM NaCl/0.05% Tween 20) or in TBST containing 1 μM 17-β estradiol. 2594 SSNHQSSRLIELLSR L-x(2)-L(2) 1 Phage display (common panning) 3 PMID:10426998 Estrogen receptor alpha, ERα Not determined. Not determined. Not determined. Not determined. In the first round, immulon 4 96-well plates (Dynex Technologies, Chantilly, VA) were coated with streptavidin in NaHCO3 buffer (pH 8.5). Wells were blocked with bovine serum albumin (BSA) and then washed with TBST [10 mM tris-HCl (pH 8.0), 150 mM NaCl, and 0.05% Tween 20], and 2 pmol of biotinylated vitellogenin ERE was then added per well. Plates were washed with TBST, 3 pmol of baculovirus-purifed ERα was then added, and plates were incubated at room temperature for 1 hour. Estradiol was then added (1 mM) along with phage library (containing about 1.5e9 phage) in TBST and incubated at room temperature for 1 hour. Affinity selection was repeated three times. 2595 SSLTSRDFGSWYASR 1 Phage display (common panning) 3 PMID:10426998 Estrogen receptor alpha, ERα Not determined. Not determined. Not determined. Not determined. In the first round, immulon 4 96-well plates (Dynex Technologies, Chantilly, VA) were coated with streptavidin in NaHCO3 buffer (pH 8.5). Wells were blocked with bovine serum albumin (BSA) and then washed with TBST [10 mM tris-HCl (pH 8.0), 150 mM NaCl, and 0.05% Tween 20], and 2 pmol of biotinylated vitellogenin ERE was then added per well. Plates were washed with TBST, 3 pmol of baculovirus-purifed ERα was then added, and plates were incubated at room temperature for 1 hour. Estradiol or tamoxifen was then added (1 mM) along with phage library (containing about 1.5e9 phage) in TBST and incubated at room temperature for 1 hour. Affinity selection was repeated three times. 2596 SSWDMHQFFWEGVSR 1 Phage display (common panning) 3 PMID:10426998 Estrogen receptor alpha, ERα Not determined. Not determined. Not determined. Not determined. In the first round, immulon 4 96-well plates (Dynex Technologies, Chantilly, VA) were coated with streptavidin in NaHCO3 buffer (pH 8.5). Wells were blocked with bovine serum albumin (BSA) and then washed with TBST [10 mM tris-HCl (pH 8.0), 150 mM NaCl, and 0.05% Tween 20], and 2 pmol of biotinylated vitellogenin ERE was then added per well. Plates were washed with TBST, 3 pmol of baculovirus-purifed ERα was then added, and plates were incubated at room temperature for 1 hour. Tamoxifen was then added (1 mM) along with phage library (containing about 1.5e9 phage) in TBST and incubated at room temperature for 1 hour. Affinity selection was repeated three times. 2597 SSISTYHMGEWFYAMLSSR SSDLYSQMREFFQINLSR SSPGSREWFKDMLSR SSTTMFDFFYERLSR SSWNSREFFLSQLSR SSVASREWWVRELSR [SM]-x-[DE]-[WF]-[WF]-x(3)-L 6 Phage display (common panning) 3 PMID:10426998 Estrogen receptor beta, ERβ Not determined. Not determined. Not determined. Not determined. In the first round, immulon 4 96-well plates (Dynex Technologies, Chantilly, VA) were coated with streptavidin in NaHCO3 buffer (pH 8.5). Wells were blocked with bovine serum albumin (BSA) and then washed with TBST [10 mM tris-HCl (pH 8.0), 150 mM NaCl, and 0.05% Tween 20], and 2 pmol of biotinylated vitellogenin ERE was then added per well. Plates were washed with TBST, 3 pmol of baculovirus-purifed ERα was then added, and plates were incubated at room temperature for 1 hour. Tamoxifen was then added (1 mM) along with phage library (containing about 1.5e9 phage) in TBST and incubated at room temperature for 1 hour. Affinity selection was repeated three times. 2598 QPLIAKWLPYLLEETVLVG NALIVPHLYELLKREWQSV 2 Phage display (common panning) 3 PMID:11306468 Estrogen receptor, ER Not determined. Not determined. Not determined. (X)7LXXLL(X)7 library 2599 SWLIDAHLAPLLFNNTLGS HFLINQHLYKLLQDTDIVV PWLDPEKLARLLEVPVQDF 3 Phage display (common panning) 3 PMID:11306468 Estrogen receptor beta, ER-beta Not determined. Not determined. Not determined. (X)7LXXLL(X)7 library 2600 CSLLNATKC CKLNANNFC CYGYLPSRC CNTSIQRNC CLPGPSHFC 5 Phage display (subtractive panning) 3/5 PMID:15200474 Tomato-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The real target is carotenoid. After deselection on cotton/selection on stained cotton swatches for two times, amplified phages were submitted to another selection or three rounds of selection on stained cotton swatches. 2601 CPYLPSIPC CYTKPYPYC CSPTTGQSC 3 Phage display (subtractive panning) 3 PMID:15200474 Tomato-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The real target is carotenoid. Phages were submitted to deselection on cotton/selection on stained cotton swatches for three times. Bound phages were eluted by the method of 'polymerase chain reaction (PCR)' elution and sequenced after the first round of biopanning. 2602 CSTSLLNAC 1 Phage display (subtractive panning) 5 PMID:15200474 Tomato-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The real target is carotenoid. After deselection on cotton/selection on stained cotton swatches for two times, amplified phages were submitted to another three rounds of biopanning on stained cotton swatches. Bound phages were eluted by the method of 'polymerase chain reaction (PCR)' elution. 2603 CGHSMLTNC CNTSYQYRC 2 Phage display (subtractive panning) 2 PMID:15200474 Tomato-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The real target is carotenoid. Phages were submitted to deselection on cotton/selection on stained cotton swatches for two times. Bound phages were eluted by the method of 'polymerase chain reaction (PCR)' elution and sequenced after the first round of biopanning. 2604 HSFRVTSNLSPP NPTTTYKMTPTM TATHLSPGAWRP SMQLQLIPSTPT 4 Phage display (subtractive panning) 3/5 PMID:15200474 Tomato-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The real target is carotenoid. After deselection on cotton/selection on stained cotton swatches for two times, amplified phages were submitted to another one round or three rounds of biopanning on stained cotton swatches. 2605 KASNLSPILGLP TVLSPLTQTLYF SLPAQPRLTHLW TVLSPLTQTLYF GTYNLPNPPPPL HYSSQSNLADSH EPTTTTLPTVGR SRNIPLPSHFLS 8 Phage display (subtractive panning) 3 PMID:15200474 Tomato-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The real target is carotenoid. Phages were submitted to deselection on cotton/selection on stained cotton swatches for three times. Bound phages were eluted by the method of 'polymerase chain reaction (PCR)' elution and sequenced after the first round of biopanning. 2606 HFAYTKPMRIPQ PWASITPPPLLR SHAITATHLEPS LQLQLLPYAFPV 4 Phage display (subtractive panning) 2 PMID:15200474 Tomato-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The real target is carotenoid. Phages were submitted to deselection on cotton/selection on stained cotton swatches for two times. Bound phages were eluted by the method of 'polymerase chain reaction (PCR)' elution and sequenced after the first round of biopanning. 2607 LNANLPANSVLA LPAQMTPVSVVR 2 Phage display (subtractive panning) 5 PMID:15200474 Tomato-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The real target is carotenoid. After deselection on cotton/selection on stained cotton swatches for two times, amplified phages were submitted to another three rounds of biopanning on stained cotton swatches. Bound phages were eluted by the method of 'polymerase chain reaction (PCR)' elution. 2608 SPLTQYVTPRHP LPAQYQTIPGSL SPLTQYVTPRHP ALGSMTWSPPPL SYTKPDQHALAF VASRLTGSVASA LPAQYQTIPGSL 7 Phage display (subtractive panning) 3/5 PMID:15200474 Paprika-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The real target is carotenoid. After deselection on cotton/selection on stained cotton swatches for two times, amplified phages were submitted to another one round or three rounds of biopanning on stained cotton swatches. 2609 SPHSMLQNPSGP YLPSTFAPPLPL 2 Phage display (subtractive panning) 4 PMID:15200474 Paprika-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The real target is carotenoid. After deselection on cotton/selection on stained cotton swatches for three times, amplified phages were submitted to another one round of biopanning on stained cotton swatches. 2610 AVSLVPPNLATH VASANPHSMTSW KAIGMSTGPLTQ VASANPHSMTSW LHVTTTIPGGLR 5 Phage display (subtractive panning) 2 PMID:15200474 Paprika-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The real target is carotenoid. Phages were submitted to deselection on cotton/selection on stained cotton swatches for two times. Bound phages were eluted by the method of 'polymerase chain reaction (PCR)' elution and sequenced after the first round of biopanning. 2611 CSNLAPMLC CNLATSNAC CSPTAAQSC 3 Phage display (subtractive panning) 4 PMID:15200474 Paprika-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The real target is carotenoid. After deselection on cotton/selection on stained cotton swatches for three times, amplified phages were submitted to another one round of biopanning on stained cotton swatches. 2612 CINTPHSMC 1 Phage display (subtractive panning) 5 PMID:15200474 Paprika-stained cotton swatches Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The real target is carotenoid. After deselection on cotton/selection on stained cotton swatches for two times, amplified phages were submitted to another three rounds of biopanning on stained cotton swatches. Bound phages were eluted by the method of 'polymerase chain reaction (PCR)' elution. 2613 DGVCQCRGDCFCP(4) DGFCECRGDCLCF(2) DGVCQCRGDCLCV(1) DGPCHCRGDCLCQ(1) DGARSCRGDCVEA(1) DGARYCRGDCFDG(1) 6 Phage display (subtractive panning) 3-4 PMID:15235124 Human dermal microvascular endothelial cells, HDMECs Not determined. Not determined. Not determined. CPL4b phage display library (DGXXXCRGDCXXX) HEK293 was used in the negative selection. 2614 DGVCQCRGDCFCP(4) DGECQCRGDCLCP(1) DGFPGCRGDCSQE(1) 3 Phage display (subtractive panning) 3-4 PMID:15235124 Human melanoma cell line MeWo Not determined. Not determined. Not determined. CPL4b phage display library (DGXXXCRGDCXXX) HEK293 was used in the nagative selection. 2615 DGALYCRGDCVDQ(4) DGARVCRGDCVDQ(2) DGEACCRGDCVCE(1) DGSRLCRGDCVDA(1) DGAFLCRGDCVDF(1) 5 Phage display (subtractive panning) 3 PMID:15235124 Mixture of murine melanoma cell lines B16.F1 and B16.F10 Not determined. Not determined. Not determined. CPL4b phage display library (DGXXXCRGDCXXX) NIH 3T3 was used in the negative selection. 2616 LSGCRGDCFQQ(1) LSACRGDCQRV(1) 2 Phage display (subtractive panning) 3 PMID:15235124 Human melanoma cell line MeWo Not determined. Not determined. Not determined. CPL4c phage display library (XXXCRGDCXXX) HEK293 was used in the nagative selection. 2617 LTAELTP(2) AASARLP(1) IVAQPRL(1) QPRLLHH(1) TRSQPPL(1) SHFSHLR(1) SPDHLFC(1) IPMHLHN(1) NHLSALY(1) ETIKTNT(1) TSFFMPP(1) SPSVVPF(1) HFTHPTH(1) SWNHARV(1) EAVPTYS(1) SDLATVR(1) LPTKTLF(1) KIDGTPR(1) WTSDELH(1) SLPRNSD(1) LCMTTLV(1) VYFPAPN(1) FPQTYTT(1) NIIPAPT(1) NTGPNRV(1) AFNYPPH(1) LEHPPTT(1) TSESPTV(1) TYSLKSA(1) ATGFATP(1) TAAYRFW(1) HSLTFSI(1) AGATAMS(1) NHWHGGL(1) INSNAPG(1) INSVSPH(1) AAWPTSS(1) VEPARAS(1) TLGLHMS(1) GYQQVFQ(1) SEVAVQG(1) FSMSTPS(1) NIAAFSL(1) SPTYPRR(1) PPPDWTF(1) DFLQVSP(1) TYPSSEW(1) YSLSRSL(1) TSTMPSR(1) TNSQPSP(1) LPPSLYS(1) WNSTTQA(1) STYPIIR(1) GILSPSH(1) YSTHSTR(1) MSSPGPA(1) SIGYPLP(1) LSNFHSS(1) QPRL, HL, PS, INS 58 Phage display (common panning) PMID:10889136 Human umbilical vein endothelial cells, HUVECs Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The peptide SIGYPLP retargeted gene delivery specifically to endothelial cells with a significantly enhanced efficiency over nontargeted adenovirus and without transduction of nontarget cells. 2618 SIGYPLP(45) MSPPGPA(33) LSNFHSS(2) 3 Phage display (subtractive panning) 4 PMID:10889136 Human umbilical vein endothelial cells, HUVECs Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Phages were biopanned on HUVECs after biopanning on 2 successive cultures of VSMCs, HepG2 cells, and peripheral blood mononuclear cells (preclearing steps). The peptide SIGYPLP retargeted gene delivery specifically to endothelial cells with a significantly enhanced efficiency over nontargeted adenovirus and without transduction of nontarget cells. 2619 LRGDWSED(1) VRGDWYEY(1) VRGDCSSS(1) GRGDLCDF(1) GRGDSPAS(1) 5 Phage display (common panning) 2 PMID:12595259 Integrin alpha-V-beta-5, integrin αvβ5 Not determined. Not determined. Not determined. JCFN-RGD phage display library (XRGDXXXX) 2620 SRGDVVPP(2) PRGDWIEF(1) GRGDSPAS(1) GRGDDDRL(1) GRGDYVLG(1) GRGDFSFL(1) GRGDWTEH(1) ARGDWVEG(1) PRGDWTEG(1) GRGDAFSL(1) FRGDSPLD(1) TRGDPPPH(1) PRGDWNEG(1) FRGDWIEL(1) 14 Phage display (common panning) 3 PMID:12595259 Integrin alpha-V-beta-5, integrin αvβ5 Not determined. Not determined. Not determined. JCFN-RGD phage display library (XRGDXXXX) 2621 CPGPEGAGC(0.14) 1 Phage display (in vivo) 4 PMID:11854520 Mouse breast tissue Not determined. Not determined. Not determined. T7 CX7C phage display library The phage-displayed peptide library was injected intravenously into mice and subsequently rescued bound phage from breast tissue. 2622 PRPGAPLAGSWPGTS(5)[24.8, 45.0] DRWRPALPVVLFPLH(1)[64.2, 36.1] ASSSYPLIHWRPWAR(1)[53.5, 29.3] AAEWLDAFFVRHVDR(1)[8.0, NT] GDVWLFLTSTSHFAR(1)[5.2, NT] EGCSVSSVGALCTHV(1)[4.8, NT] APCCSHLDASPFQRP(1)[3.6, NT] PAQSNFVTWGYNVAV(1)[2.8, NT] RASDVGSDVVPRYPF(1)[1.5, NT] 9 Phage display (in vivo) 5 PMID:11965539 Angiogenic vessel Not determined. Not determined. Not determined. X15 fUSE5 phage display library Binding assay For affinity screening, B16BL6 cells were implanted subcutaneously into the posterior fank of 5-week-old C57BL/6 male mice. Each sample of phage clones (1.0e11 c.f.u.) was injected into tumor-bearing mice via a tail vein when the tumor size had become about 10 mm in diameter. Four minutes after injection, the phages that had accumulated in tumor tissue were recovered and titrated. Phage affinity was expressed as a fold of the control value obtained with random peptide expressing-phage. Both phage affinity against B16BL6 melanoma and phage affinity against Meth A sarcoma are shown. NT denotes not tested. The phage-displayed peptide library was injected into DAS model mice via a tail vein. Four minutes after injection, the phages that had accumulated in angiogenic vessels were recovered and titrated. Biopanning steps were repeated for five cycles. 2623 GECRENVCMG(10) NTCKRFNCNL(2) 2 Phage display (competitive panning) 3 PMID:9288781 Angiogenin Actin Not determined. Not determined. XXCX4CXX phage display library In the first round, bound phages were eluted by a high concentration of actin (5 μM) following washing with a lower concentration of actin (50 nM). In the second round, bound phages were first eluted with 50 nM actin and then by glycine-HCI, pH 2.2. In the third round, bound phages were eluted by a high concentration of actin (5 μM) following washings with lower concentrations of actin (first 50 nM, second 500 nM). 2624 GECRENVCMG(13/15) 1 Phage display (competitive panning) 3 PMID:9288781 Angiogenin Actin Not determined. Not determined. XXCX4CXX phage display library In the first round, bound phages were eluted by a high concentration of actin (5 μM) following washing with a lower concentration of actin (50 nM). In the second round, bound phages were first eluted with 50 nM actin and then by glycine-HCI, pH 2.2. In the third round, bound phages were eluted by a high concentration of actin (5 μM) following washings with lower concentrations of actin (first 50 nM, second 500 nM) and subsequenctly eluted with glycine-HCI, pH 2.2. After three rounds of panning, 15 phages were harvested. Thirteen clones encode GECRENVCMG, except for two clones that did not yield any identifiable sequence. 2625 VVACSWDWTMGAVVCYERI(1/25) WTGWCLNPEESTWGFCTGSF(1/25) 2 Phage display (subtractive panning) 1 PMID:15300809 Extracellular domain of receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Not determined. X4CX10CX4 fUSE5 phage display library In the first round, 0.9 μg ECDAP were incubated with anti-AP conjugated agarose beads on a nutator overnight at 4°C. The amplified phage library was incubated with anti-AP conjugated agarose beads alone for 2 hr on a nutator at room temperature to remove those phage specific for AP. The unbound phage were filtered with a 0.45 μm cellulose acetate syringe filter and incubated with ECDAP bound to beads on a nutator for 2 hr at room temperature. The unbound phage were removed by washing 5 times with PBS containing a cocktail of prokaryotic protease inhibitors (PPI) (Sigma). One clone (EC-1, displaying WTGWCLNPEESTWGFCTGSF) bound all preparations of ErbB2 including live cells and fresh frozen human breast cancer specimens. The synthetic peptide based on the deduced sequence of the EC-1 clone and its biotin-conjugated form retained binding affinity for purified ErbB2 and ErbB2 overexpressing cell lysates. EC-1 peptide was able to effectively inhibit the phosphorylation of ErbB2 on residues Y1248 and Y877 in a dose- and time-dependent manner. Furthermore, EC-1 peptide selectively inhibits the proliferation of ErbB2 overexpressing breast cancer cells. The linear portion of the cyclic EC-1 peptide was shown to be essential for binding ErbB2. In addition, 4 biased phage libraries were constructed allowing 4 different regions of the EC-1 peptide to have random sequence. 2626 VVACSWDWTMGAVVCYERI(1/39) 1 Phage display (subtractive panning) 3 PMID:15300809 Extracellular domain of receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Not determined. X4CX10CX4 fUSE5 phage display library In the first round, 0.9 μg ECDAP were incubated with anti-AP conjugated agarose beads on a nutator overnight at 4°C. The amplified phage library was incubated with anti-AP conjugated agarose beads alone for 2 hr on a nutator at room temperature to remove those phage specific for AP. The unbound phage were filtered with a 0.45 μm cellulose acetate syringe filter and incubated with ECDAP bound to beads on a nutator for 2 hr at room temperature. The unbound phage were removed by washing 5 times with PBS containing a cocktail of prokaryotic protease inhibitors (PPI) (Sigma). Additional two rounds of screening were performed. 2627 WTGWCLNPEESTWGFCTGSF(9/25) LNWECWYDYRLEAWDCRGDI(4/25) DTDMCWWWSREFGWECAGAG(2/25) VVACSWDWTMGAVVCYERI(1/25) 4 Phage display (subtractive panning) 3 PMID:15300809 Extracellular domain of receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Not determined. X4CX10CX4 fUSE5 phage display library In the first round, 0.9 μg ECDAP were incubated with anti-AP conjugated agarose beads on a nutator overnight at 4°C. The amplified phage library was incubated with anti-AP conjugated agarose beads alone for 2 hr on a nutator at room temperature to remove those phage specific for AP. The unbound phage were filtered with a 0.45 μm cellulose acetate syringe filter and incubated with ECDAP bound to beads on a nutator for 2 hr at room temperature. The unbound phage were removed by washing 5 times with PBS containing a cocktail of prokaryotic protease inhibitors (PPI) (Sigma). Additional two rounds of screening were performed. One clone (EC-1, displaying WTGWCLNPEESTWGFCTGSF) bound all preparations of ErbB2 including live cells and fresh frozen human breast cancer specimens. The synthetic peptide based on the deduced sequence of the EC-1 clone and its biotin-conjugated form retained binding affinity for purified ErbB2 and ErbB2 overexpressing cell lysates. EC-1 peptide was able to effectively inhibit the phosphorylation of ErbB2 on residues Y1248 and Y877 in a dose- and time-dependent manner. Furthermore, EC-1 peptide selectively inhibits the proliferation of ErbB2 overexpressing breast cancer cells. The linear portion of the cyclic EC-1 peptide was shown to be essential for binding ErbB2. In addition, 4 biased phage libraries were constructed allowing 4 different regions of the EC-1 peptide to have random sequence. 2628 VVACSWDWTMGAVVCYERI(1/25) WTGWCLNPEESTWGFCTGSF(1/25) 2 Phage display (subtractive panning) 1 PMID:15300809 Extracellular domain of receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Not determined. X4CX10CX4 fUSE5 phage display library In the first round, 20 μg CBECD was incubated in a polystyrene tube overnight at 4°C on a nutator. The isolated phage were pre-incubated in a casein blocked polystyrene tube to remove peptide-phage that bound to casein or polystyrene. The pre-cleared phage not bound to the casein/polystyrene tube were then added to the tube coated with CBECD and incubated on a nutator for 2 hr at room temperature. The non-bound phage were washed from beads by washing 5 times with PBS containing PPI. One clone (EC-1, displaying WTGWCLNPEESTWGFCTGSF) bound all preparations of ErbB2 including live cells and fresh frozen human breast cancer specimens. The synthetic peptide based on the deduced sequence of the EC-1 clone and its biotin-conjugated form retained binding affinity for purified ErbB2 and ErbB2 overexpressing cell lysates. EC-1 peptide was able to effectively inhibit the phosphorylation of ErbB2 on residues Y1248 and Y877 in a dose- and time-dependent manner. Furthermore, EC-1 peptide selectively inhibits the proliferation of ErbB2 overexpressing breast cancer cells. The linear portion of the cyclic EC-1 peptide was shown to be essential for binding ErbB2. In addition, 4 biased phage libraries were constructed allowing 4 different regions of the EC-1 peptide to have random sequence. 2629 WTGWCLNPEESTWGFCTGSF(8/25) LNWECWYDYRLEAWDCRGDI(4/25) DTDMCWWWSREFGWECAGAG(2/25) 3 Phage display (subtractive panning) 2 PMID:15300809 Extracellular domain of receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Not determined. X4CX10CX4 fUSE5 phage display library In the first round, 20 μg CBECD was incubated in a polystyrene tube overnight at 4°C on a nutator. The isolated phage were pre-incubated in a casein blocked polystyrene tube to remove peptide-phage that bound to casein or polystyrene. The pre-cleared phage not bound to the casein/polystyrene tube were then added to the tube coated with CBECD and incubated on a nutator for 2 hr at room temperature. The non-bound phage were washed from beads by washing 5 times with PBS containing PPI. Another one round of screening was performed. One clone (EC-1, displaying WTGWCLNPEESTWGFCTGSF) bound all preparations of ErbB2 including live cells and fresh frozen human breast cancer specimens. The synthetic peptide based on the deduced sequence of the EC-1 clone and its biotin-conjugated form retained binding affinity for purified ErbB2 and ErbB2 overexpressing cell lysates. EC-1 peptide was able to effectively inhibit the phosphorylation of ErbB2 on residues Y1248 and Y877 in a dose- and time-dependent manner. Furthermore, EC-1 peptide selectively inhibits the proliferation of ErbB2 overexpressing breast cancer cells. The linear portion of the cyclic EC-1 peptide was shown to be essential for binding ErbB2. In addition, 4 biased phage libraries were constructed allowing 4 different regions of the EC-1 peptide to have random sequence. 2630 VPQQSIP RGDVPP VRGRGSK NPGSSLG PGRGDL SRKDV QRSGRGP LPNRSQI VLGRGK DLISSK 10 Phage display (subtractive panning) 3 PMID:15569135 IA/IAA-enriched immunoglobulin G from REM-A Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) After this first round of‘positive' selection with IAA sera, 'negative' selection was achieved whereby phages not specifically reactive with IAA sera were removed using 100 μg of IgG prepared from pooled normal sera. The amplified eluate was enriched by two further rounds of positive selection, negative selection, and amplification. 2631 LDQRSSK PGSRLQP LQPISN VIQGSRL QVRGGS NIKSSLP PDGSRLV QQSPPI VQLNPD ARSKPN 10 Phage display (subtractive panning) 3 PMID:15569135 IA/IAA-enriched immunoglobulin G from REM-B Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) After this first round of‘positive' selection with IAA sera, 'negative' selection was achieved whereby phages not specifically reactive with IAA sera were removed using 100 μg of IgG prepared from pooled normal sera. The amplified eluate was enriched by two further rounds of positive selection, negative selection, and amplification. 2632 VPQQSIP RGDVPP VGERGSK NPGSSLG GERAKRS STGERIQ QGERSGP LPGERQI DLISSK VLGERK 10 Phage display (subtractive panning) 3 PMID:15569135 IA/IAA-enriched immunoglobulin G from NREM-A Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) After this first round of‘positive' selection with IAA sera, 'negative' selection was achieved whereby phages not specifically reactive with IAA sera were removed using 100 μg of IgG prepared from pooled normal sera. The amplified eluate was enriched by two further rounds of positive selection, negative selection, and amplification. 2633 LDQSRLK PGTRLQP PSRLVNQ VIQGSQL QVRGGS NIKSSLP LQPISN QQSRLI VSRSLLD ARSKPN 10 Phage display (subtractive panning) 3 PMID:15569135 IA/IAA-enriched immunoglobulin G from NREM-B Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) After this first round of‘positive' selection with IAA sera, 'negative' selection was achieved whereby phages not specifically reactive with IAA sera were removed using 100 μg of IgG prepared from pooled normal sera. The amplified eluate was enriched by two further rounds of positive selection, negative selection, and amplification. 2634 AKWSDLWLAAPG EQNSYWKQLFLE HHMKDFATLFST NTNAFSRLFYPS HKFQELYQSTTP DHSKLYSLLQSS [FW]-x(2)-L-[FW], F-x(2)-L-Y 6 Phage display (common panning) 3 PMID:12714604 RAR-DBD-LBD Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Each round of panning was performed on streptavidin-coated wells of a 96-well enzyme-linked immunosorbent assay plate. 2635 TATPGTWYNLWL EPNSYWKQLFLE TRQPPTTFYSLF QNFKSLFSATLP SSQRFVDLFSND HTPTTFSRLWTP DRDPSKFSLLWH QPKHFTELYFKS TSRFSHFQELYS STTKFGTLYWEN [FW]-x(2)-L-[FW] 10 Phage display (common panning) 4 PMID:12714604 RAR-DBD-LBD Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2636 TGGGVSLLLHLLNTEQGES RRDDFPLLISLLKDGALSQ YGLKMSLLESLLREDISTV MSYDMLSLYPLLTNSLLEV FPAEFPLLTYLLERQGMDE VESEFPYLLSLLGEVSPQP 6 Phage display (common panning) 3 PMID:11117531 Estrogen receptor beta, ER-beta Not determined. Not determined. Not determined. (X)7LXXLL(X)7 library Each round of panning was performed in the absence of 17β-estradiol. 2637 VSSEGRLLIDLLVDGQQSE DTPQSPLLWGLLSSDRVEG GGTQDGYLWSLLTGMPEVS SLPEEGFLMKLLTLEGDAE VMGNNPILVSLLEEPSEEP VLVEHPILGGLLSTRVDSS QTPLLEQLLTEHIQQG 7 Phage display (common panning) 3 PMID:11117531 Estrogen receptor beta, ER-beta Not determined. Not determined. Not determined. (X)7LXXLL(X)7 library Each round of panning was performed in the presence of 17β-estradiol. 2638 EDGGSLLRWYLEHEFGGGGSGK EDGGSPLWQIFSRELGGGGSK EDGGSWVEWVEEERRGGGGK EDGGSRRFTFQYGSV EDGGSLFERVWLREL 5 Phage display (common panning) 2-3 PMID:14976226 Estrogen receptor, ER Not determined. Not determined. Not determined. fGWX10 phage display library Biotinylated ERα LBD or biotinylated ERα FL was immobilized on a neutravidin-coated microtiter plate well. Each round od panning was performed on neutravidin-coated microtiter plate well. 2639 EYHEKRWLEGHIHHRIKSLLENS(16) QETIQRWLRGHIQRELGTMELKD(14) HSTTLTGLASIIRERILTELRDE(1) PENFRQALRAHIADLITNQDYRS(1) 4 Phage display (common panning) 3 PMID:12145334 Vitellogenin estrogen response element (ERE) Not determined. Not determined. Not determined. X7LXX[HI]IXXX[IL]X7 CoRNR box phage display library The target was biotinylated. Each round of panning was performed on streptavidin-coated Immulon 4 plates. 2640 ELFDAFQLRQLILRGLQDDIPYH(16) EYHEKRWLEGHIHHRIKSLLENS(1) 2 Phage display (common panning) 3 PMID:12145334 Estrogen receptor beta, ER-beta Not determined. Not determined. Not determined. X7LXX[HI]IXXX[IL]X7 CoRNR box phage display library The real target is ERβ treated with 4-hydroxytamoxifen. The target was biotinylated. Each round of panning was performed on streptavidin-coated Immulon 4 plates. 2641 EMEWMKALRQHISGELRRNYTEE(5) AAYDPAALNNNIRYALVKHSQIK(3) KWESLDALQGLISSHLSAMGPIP(2) FMNNGVLLATNIENLLRTQPGSN(1) NAAPRTALSHHIHSDLLDGPTTT(1) PKTPGVPLNPLISPEITSDTSML(1) NTYNTGALRFNIVESIWASKKLR(1) VPRFTKGLVGNIPLAIDTNSGTV(1) ESERANLLKEHIKMTLPEERKKT(1) NVIAQPTLASIIPPSLKRQSEAR(1) SPFTQVTLKGNIAPSIVGSQGMA(1) PIAHRVNLRHNITEDITLSHRFL(1) IDDGHPHLWKNIWDTLDKPGLGA(1) FKPGTSSLDTHIPLGLNKSFHHN(1) NQGNTKELLGNINYFLTHHTVPA(1) LTYPIRELKMNITSGIRLDKRVL(1) HKDSQLTLANNIMGQLSSTGGKH(1) WDVGEIRLRRHIKMPLSEEAIAE(1) SHYEKYSLPGIIVRKISTTDWRP(1) QHQNRQQLGSNIAATLPGKRESV(1) KSQTKAELPYLIGKQITKNQPEQ(1) DALDNQRLLGNIDNVLKVTNPNA(1) GQSARFALSQHIPSKIYDHPRPN(1) RAVHERALNPLILDRLLHELKSE(1) 24 Phage display (common panning) 3 PMID:12145334 Estrogen receptor beta, ER-beta Not determined. Not determined. Not determined. X7LXX[HI]IXXX[IL]X7 CoRNR box phage display library The real target is ERβ treated with ICI 182, 780. The target was biotinylated. Each round of panning was performed on streptavidin-coated Immulon 4 plates. 2642 DWEYSVWLSN(23) STEHSEADLW(8) LYFEDYRCEL(3) DWDYGALMWA(1) YSDWDYSEGL(1) [DE]-W-[DE]-Y-[SG] 5 Phage display (common panning) 3/4 PMID:9050886 Anti-dsDNA monoclonal antibody R4A Not determined. Not determined. Not determined. L100 phage display library (X10) For the first round of selection, 50 μl of streptavidin-coated magnetic beads were incubated with 100 μl of 5 μM biotinylated protein G for 1 hr at room temperature with agitation. In subsequent rounds of selection, 10 μl of streptavidin beads and 0.05 μM of biotinylated protein G were used. The streptavidin bead/biotinylated protein G complex was combined with the phage/antibody mixture and incubated. 2643 WCEADYGRCP(23) FSDCYHSGCP(3) VPVCDWELNC(1) 3 Phage display (common panning) 3/4 PMID:9050886 Anti-dsDNA monoclonal antibody R4A Not determined. Not determined. Not determined. L100 phage display library (X10) For the first round of selection, 50 μl of streptavidin-coated magnetic beads were incubated with 100 μl of 5 μM biotinylated protein G for 1 hr at room temperature with agitation. In subsequent rounds of selection, 10 μl of streptavidin beads and 0.05 μM of biotinylated protein G were used. The streptavidin bead/biotinylated protein G complex was combined with the phage/antibody mixture and incubated. 2644 SLDELDWDSM(4) RHEDGDWPRV(1) LLDDGFWPRV(1) CGVDGRWPRW(1) SLISDEWPRW(1) DGEWPREGWS(1) EDLEGEWPMR(1) [DE]-G-[DE]-W-P-R 7 Phage display (common panning) 3/4 PMID:9050886 Anti-dsDNA monoclonal antibody 52b3 Not determined. Not determined. Not determined. L100 phage display library (X10) For the first round of selection, 50 μl of streptavidin-coated magnetic beads were incubated with 100 μl of 5 μM biotinylated protein G for 1 hr at room temperature with agitation. In subsequent rounds of selection, 10 μl of streptavidin beads and 0.05 μM of biotinylated protein G were used. The streptavidin bead/biotinylated protein G complex was combined with the phage/antibody mixture and incubated. 2645 HIPAYATHV[1.40] EHFWEQRPR[1.11] WLVQSPPW[1.14] QSHFLLQGT[1.12] KRHFLSQRQ[1.23] HFLSQNFFG[1.66] WGPFQYAAG[1.98] LLRQARERP[0.96] PPLSQRRAL[1.46] TRQQNNPER[0.90] 10 Phage display (common panning) 2 PMID:9368618 Anti-the 0-Ag of the S. flexneri serotype 5a LPS monoclonal antibody C5 O-antigen of lipopolysaccharide, O-Ag of LPS Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA Wild-type phage (pwt) (cross-reacting with the Shigeffa LPS core moiety) was used as the negative control and its absorbance value is 0.05. The absorbance is shown. In the first round, the mAb was incubated at 1 μM concentration overnight at 4°C with 1.0e10 Amp(R) tranducing unit (TU) of library in a total volume of 10 pl. The mixture was incubated with 0.25 μg of a biotin-conjugated goat antimouse IgA secondary antibody ((α-chain specific), which was previously pre-adsorbed overnight at 4°C with 2.0e11 phage particles of UV-killed M13K07 to prevent nonspecific binding, and then the phage-mAb-secondary Ab complexes were tethered on streptavidin-coated dishes. The second round of affinity selection was carried out in the same way, but using 10 nM or 0.1 nM concentrations of mAb and proportionally lower amounts of the secondary antibody. 2646 KVPAWARRL[0.18] KVPPWARTA[0.51] 2 Phage display (common panning) 2 PMID:9368618 Anti-the 0-Ag of the S. flexneri serotype 5a LPS monoclonal antibody I3 O-antigen of lipopolysaccharide, O-Ag of LPS Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA Wild-type phage (pwt) (cross-reacting with the Shigeffa LPS core moiety) was used as the negative control and its absorbance value is 0.05. The absorbance is shown. In the first round, the mAb was incubated at 1 μM concentration overnight at 4°C with 1.0e10 Amp(R) tranducing unit (TU) of library in a total volume of 10 pl. The mixture was incubated with 0.25 μg of a biotin-conjugated goat antimouse IgA secondary antibody ((α-chain specific), which was previously pre-adsorbed overnight at 4°C with 2.0e11 phage particles of UV-killed M13K07 to prevent nonspecific binding, and then the phage-mAb-secondary Ab complexes were tethered on streptavidin-coated dishes. The second round of affinity selection was carried out in the same way, but using 10 nM or 0.1 nM concentrations of mAb and proportionally lower amounts of the secondary antibody. 2647 CGSPLRQRRSC[0.20] CGSPLRQRSLC[0.19] CSQGRWPWURC[0.31] CYKPLGALTHC[0.85] 4 Phage display (common panning) 2 PMID:9368618 Anti-the 0-Ag of the S. flexneri serotype 5a LPS monoclonal antibody I3 O-antigen of lipopolysaccharide, O-Ag of LPS Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA Wild-type phage (pwt) (cross-reacting with the Shigeffa LPS core moiety) was used as the negative control and its absorbance value is 0.05. The absorbance is shown. In the first round, the mAb was incubated at 1 μM concentration overnight at 4°C with 1.0e10 Amp(R) tranducing unit (TU) of library in a total volume of 10 pl. The mixture was incubated with 0.25 μg of a biotin-conjugated goat antimouse IgA secondary antibody ((α-chain specific), which was previously pre-adsorbed overnight at 4°C with 2.0e11 phage particles of UV-killed M13K07 to prevent nonspecific binding, and then the phage-mAb-secondary Ab complexes were tethered on streptavidin-coated dishes. The second round of affinity selection was carried out in the same way, but using 10 nM or 0.1 nM concentrations of mAb and proportionally lower amounts of the secondary antibody. 2648 CTRGHFLQNRC[1.79] CRRHFLEQRGC[1.92] CSPHFFNQIRC[1.26] 3 Phage display (common panning) 2 PMID:9368618 Anti-the 0-Ag of the S. flexneri serotype 5a LPS monoclonal antibody C5 O-antigen of lipopolysaccharide, O-Ag of LPS Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) ELISA Wild-type phage (pwt) (cross-reacting with the Shigeffa LPS core moiety) was used as the negative control and its absorbance value is 0.05. The absorbance is shown. In the first round, the mAb was incubated at 1 μM concentration overnight at 4°C with 1.0e10 Amp(R) tranducing unit (TU) of library in a total volume of 10 pl. The mixture was incubated with 0.25 μg of a biotin-conjugated goat antimouse IgA secondary antibody ((α-chain specific), which was previously pre-adsorbed overnight at 4°C with 2.0e11 phage particles of UV-killed M13K07 to prevent nonspecific binding, and then the phage-mAb-secondary Ab complexes were tethered on streptavidin-coated dishes. The second round of affinity selection was carried out in the same way, but using 10 nM or 0.1 nM concentrations of mAb and proportionally lower amounts of the secondary antibody. 2649 YRNQYQPPPYASSQ LQRDRPPPPYPTP SYYRQPPPPYPTSA SQRGTPPPPYPFAA SRQVLEPFYPQKA RCWWGPPPPYPKVA LMRIDPPPPYPSM WWNYDEPPAYWSTI PWVLEPPPPYPGT RAAGRLPPPYWELE WQRCTSPPAYYDAI EWKREPPPEYRPPP x-P-P-x-Y 12 Phage display (common panning) 3 PMID:9224934 Yorkie homolog Not determined. Not determined. Not determined. SSX6PPX6SR M13 phage display library 2650 DQLRDEPPAYWDVV QMEREPPPPYPQPS GWGEVPPPPYRPKE RDGYGPPPPYRPPT RAESGSPPPYPLES ASAKIMPPPYPSNN WLRRGIPPPYPSTE GLYQGPPPTYPTR SHERIPPPPYPHS RISRYPPPAYNTDN RGELSNPPPYPPLR SSRQAPPPPYPRDY x-P-P-x-Y 12 Phage display (common panning) 3 PMID:9224934 Yorkie homolog Not determined. Not determined. Not determined. SSX6PPX6SR M13 phage display library 2651 GLHSRCHIGRDCSSA(9/21) GFVCSGIFAVGVGRC(7/21) HYGDCRYDLGSCRGA(2/21) APGVRLGCAVLGRYC(1/21) ACVMYDFVLRGMCAR(1/21) 5 Phage display (common panning) 3 PMID:9528007 Anti-CTLA4 monoclonal antibody Not determined. Not determined. Not determined. X15 fUSE5 phage display library 2652 CKGGRAKDC CVMGSVTGC CRGEVLWSC 3 Phage display (subtractive panning, in vivo) 4 PMID:15133506 White fat Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In each biopanning round, an adult obese leptin-deficient [Lep(ob/ob)] female mouse was injected intravenously through the tail vein with 1.0e10 transducing units (TU) of the library. Phage (～300 TU/g in round 1 increased to ～e4 TU/g in round 3) were recovered after 5 min of circulation from subcutaneous fat and bulk-amplified for each subsequent round. Phage amplified after the third round of panning were enriched for fat-specific binders by adapting an in vivo subtraction step: a lean, C57BL/6 female was injected intravenously with 1.0e9 TU of phage selected in round 3. After 5 min, the unbound phage were recovered from circulation and amplified for the fourth and final round of biopanning in a Lep(ob/ob) female mouse. 2653 CYPLRCDYSSAAPC 1 Phage display (competitive panning) 2 PMID:9387141 Angiogenin Not determined. Not determined. Not determined. CX12C phage display library The phages bound specifically to the actin-binding site of angiogenin were competitively eluted with bovine actin. 2654 CRGDGWMC(10) CRGDGWC(8) CRGDGLMC(7) CRGDGWLC(5) CRGDGMLC(5) CRGDGFC(3) CRGDGLMCGL(2) CRGDGMLC(1) CRGDGWIC(1) CRGDGWWC(1) CRGDGLIC(1) CRGDGLDC(1) CRGDGLLC(1) CRGDGLWC(1) CRGDGFLC(1) CRGDGQHC(1) CRGDGAFC(1) CRGDGAWC(1) CRGDNVWC(1) CRGDNAWC(1) CRGDAAWC(1) CRGDAAHC(1) CRGDRAWC(1) CPVRRGDGWC(1) CDWRGDNQFC(1) CLRGDGLALC(1) CWRGDHVMPC(1) CGQRGDGFCL(1) CRGDGYCVFF(1) CRGDCLPTPR(1) CYVNGRAWAC(1) CTNVNGRSAC(1) CQGMHGTPAC(1) CQGIDGTPAC(1) CGQGMHRLAC(1) CMWLSVNYSC(1) CREQPASRSC(1) CKWRSARDLC(1) CVDCILRYLC(1) CGADSEEGPC(1) RGD 40 Phage display (common panning) 2/3/4 PMID:9634769 Integrin alpha-5-beta-1, integrin α5β1 Fibronectin, FN Not determined. Not determined. CX5C+CX6C+CX9 FUSE5-based phage display library pool 2655 CWRGDTPC(1) CWRGDRAC(1) CLRGDRVC(1) CSRGDGRC(1) CRGDSLRC(1) CRGDGRNC(1) CTTRGDSFC(1) CRVRGDSWC(1) CLRRGDSGC(1) CISRGDTFC(1) CPSRGDALC(1) CAGRGDALC(1) CSPRGDAGC(1) CTRRGDATC(1) CVRRGDAFC(1) CLSRGDVVC(1) CNARGDGFC(1) CVTRGDHFC(1) CEVRGDRIC(1) CNIRGDKIC(1) CNARGDKLC(1) CPRGDSTLC(1) CTRGDSIFC(1) CTRGDSLDC(1) CGRGDSHHC(1) CDRGDSQSC(1) CSRGDTYLC(1) CLRGDIANC(1) CGRGDLIHC(1) CSRGDGAIC(1) CFRGDDRKC(1) CRGDSFVGC(1) CRGDSHLQC(1) CRGDNTFGC(1) CRGDTVYAC(1) CRGDHGTLC(1) CRGDAWPGC(1) CRGDLAWVC(1) CRGDGIRFC(1) CRGDKGWNC(1) CARRLDAPC(1) CPSRLDSPC(1) CKTPGRLDC(1) CTTRSDSFC(1) CWSISPYFC(1) CPDLLAQSC(1) CLVLPSTGC(1) CYSLGFLVC(1) CQARGDRPRC(1) CNRRGDNWGC(1) RGD 50 Phage display (common panning) 2/3/4 PMID:9634769 Integrin alpha-V-beta-3, integrin αvβ3 Fibronectin, FN Not determined. Not determined. CX5C+CX6C+CX7C+CX9 phage display library pool 2656 CRRGDFGGC(2) CYARGDYPC(2) CSGRHDDYC(2) CFSRGDFPC(1) CHIRGDFPC(1) CRYRGDLPC(1) CSARGDWPC(1) CKRGDWIRC(1) CGARGDSRC(1) CRRMDMPDC(1) CWARRDMPC(1) CWVRSDLGC(1) CPLRRDWIC(1) CTARSDRRC(1) CMSRADRPC(1) CHSTRDELC(1) CRGDNYWC(1) CRGDNSAC(1) CPRGDWPC(1) CGRGDQLC(1) CVRGDRMC(1) CRGDRALC(1) CRGDTRSC(1) CGRGDGDC(1) CRQDVPQC(1) CRADVPLC(1) CGRLDVPC(1) CYRRDVPC(1) CKGDMPRC(1) CRHDSPRC(1) CKRRDYPC(1) CTRTDGWC(1) CMRTDGRC(1) CRTRDSPC(1) CVVRDMPC(1) RGD 35 Phage display (common panning) 2/3/4 PMID:9634769 Integrin alpha-M-beta-3, integrin αmβ3 Fibrinogen alpha, beta and gamma chain 2VDO,2VDP,2VDQ,2VDR, Not determined. CX5C+CX6C+CX7C FUSE5 phage display library pool 2657 CVTRGDNFC(2) CDCRGDCFC(2) CDCRGDCLC(2) CIRRGDTFGC(2) CAPRGDHFAC(2) CRGDTFC(1) CRGDVFLC(1) CWTRGDSFC(1) CEGRGDSFC(1) CYARGDSFC(1) CEPRGDSFC(1) CELRGDSAC(1) CLVRGDSLC(1) CHTRGDTFC(1) CISRGDTFC(1) CVVRGDTFC(1) CEMRGDTFC(1) CDLRGDTYC(1) CVLRGDNFC(1) CVRRGDVFC(1) CGRGDTPTC(1) CRGDTYLIC(1) CLCRGDCIC(1) CRGDCCQSC(1) CLHPNVRSC(1) CDSVLRVFC(1) CQGRGDTFYC(1) CPRRGDTFSC(1) CAHRGDTPQC(1) CVSRGDTPKC(1) CVTRGDSFSC(1) CQVRGDQFAC(1) CTQRGDNFFC(1) CQSRGDDFSC(1) CGRRGDVPRC(1) CRGDTPGFLC(1) CRGDLPRAWC(1) CRGDVPAVGC(1) CYRGDADFWC(1) CSQKRGDTWC(1) CPDRGDKTYC(1) CGPRERFLSC(1) CIRQRIYPWC(1) CGQRSSARAS(1) CGSPLKSIKC(1) CIEIQHGKAC(1) CLESRGPQKC(1) CRKQVMACTA(1) CEAKFQLHWV(1) CVGKELHKRV(1) CTRKRAVGAA(1) RGD 51 Phage display (common panning) 2/3/4 PMID:9634769 Integrin alpha-V-beta-5, integrin αvβ5 Not determined. Not determined. Not determined. CX5C+CX6C+CX7C+CX9 phage display library pool 2658 EFKHSVVGCEALLRYNTPPDPELIC(1) CPRHSIVEAAALLRY(1) SRHSVLEPALLRYSEIQAQSFHG(1) CPRHSIVETAALLRY(1) GRHSVLGPSMALLRY(1) AELRHSVMLGALLRYGAIEPRGKSH(1) SRHSVLAPALLRYEEIIAYSGSS(1) AASTSPGIGAALLRYYQIKKHSLIP(1) SRHSVLAPALLRY(1) VFRHSLVWSQALLRY(1) RHSMVSVDVRALLRY(1) TRHSILRPVNALLRYCIETNNDC(1) SSLSDASMVDALLRYYQIKKHSLIP(1) VHADLHDNTKALLRYYQIKKHSLIP(1) EFKHSVVGCEALLRYHVTHGLT(1) CIMRHSVVPDALLRYAPEDQRQICH(1) VFRHSLVWSQALLRYAPEDQRQICH(1) GRHSVLGPSMALLRY(1) SRHSVLEPALLRYSIIEAHSGTC(1) EFKHSVVGCEALLRYTRSALTSDPC(1) EFKHSVVGCEALLRYGGIAPHRCTC(1) 21 Phage display (common panning) 4 PMID:8755549 Anti-P53 monoclonal antibody PAb240 Not determined. Not determined. Not determined. X10ALLRYX10 phage display library 2659 SISFGQLWRPALLRYTEASNIIRRT(2) SISFGQLWRPALLRLLHKDNAFVR(1) SISFGQLWRPALLRYESHAGSPRGR(1) SISFGQLWRPALLRYTDAVNTSLRI(1) SISFGQLWRPALLRYDHSTTARINL(1) EVRWFDWIHKALLRYPTVLINVRNP(1) SISFGQLWRPALLRYEISPASVRLR(1) 7 Phage display (common panning) 3 PMID:8755549 Alkaline phosphatase, tissue-nonspecific isozyme Not determined. Not determined. Not determined. X10ALLRYX10 phage display library 2660 SISFGQLWRPALLRYGSTPVTLAIS(4) SISFGQLWRPALLRYDVKILHGSQR(2) 2 Phage display (common panning) 4 PMID:8755549 Alkaline phosphatase, tissue-nonspecific isozyme Not determined. Not determined. Not determined. X10ALLRYX10 phage display library 2661 DLMGLRNSVLALLRYKLPKPTPGPN(19) 1 Phage display (common panning) 3-4 PMID:8755549 Alkaline phosphatase, tissue-nonspecific isozyme Not determined. Not determined. Not determined. X10ALLRYX10 phage display library 2662 WYEYGWDETVALLRYIEGTSISNAA(3) 1 Phage display (common panning) 3-4 PMID:8755549 Beta-glucuronidase, GUS Not determined. Not determined. Not determined. X10ALLRYX10 phage display library 2663 DPVTDEWVWEALLRYQQATARILLS(3) 1 Phage display (common panning) 3 PMID:8755549 Beta-glucuronidase, GUS Not determined. Not determined. Not determined. X10ALLRYX10 phage display library 2664 DPLSAFGWNAALLRYKHDIQTVYAQ(4) DPVFYVDVLPALLRYQKIFANNI(3) DPVFYVDVLPALLRYTGSTIPTTIR(2) 3 Phage display (common panning) 4 PMID:8755549 Beta-glucuronidase, GUS Not determined. Not determined. Not determined. X10ALLRYX10 phage display library 2665 CRNPWC 1 Phage display (common panning) PMID:9050835 BCL1 IgM idiotype Not determined. Not determined. Not determined. f3-6mer phage display library (X6) 2666 TAAGLCEFDQ 1 Phage display (common panning) PMID:9050835 BCL1 IgM idiotype Not determined. Not determined. Not determined. X10ALLRYX10 phage display library 2667 MSRPACPNDKYE MSARHVPNDKYE MTSRTSPNDKYE MSRSDVPNDKYE GSRVPWPNDKYE GARLPYPNDKYE SAAARVPNDKYE SQVCRRPNDKYE SFVWRMPNDKYE RMAKPVPNDKYE MSQRIQPNDKYE MTHPVAPNDKYE MHAGELPNDKYE MMIGTNPNDKYE MMISPNDKYE GSRGDWPNDKYE 16 Phage display (common panning) 3 PMID:7990127 Thrombin receptor Not determined. Not determined. Not determined. X6PNDKYEPF phage display library Platelets were mixed with phage particles. After several cycles of washing, the resulting platelet/phage pellet was resuspended in PBS, milk containing SFLLRNPNDKYE (thrombin receptor activating peptide). 2668 ADGAARPIAPGAAG ADGAARTVSPGAAG ADGAARVPSTGAAG ADGASRAFSYGAAG ADGAARMPSAGAAG ADGAARVFLRGAAG ADGASRPVYVGAAG ADGASRMVHVGAAG 8 Phage display (common panning) 3 PMID:7990127 Thrombin receptor Not determined. Not determined. Not determined. f3-6mer phage display library (X6) The real target is thrombin receptor on platelets. Platelets were mixed with phage particles. After several cycles of washing, the resulting platelet/phage pellet was resuspended in PBS, milk containing SFLLRNPNDKYE (thrombin receptor activating peptide). 2669 SVWRWLPYDKYE 1 Phage display (common panning) PMID:9050835 BCL1 IgM idiotype Not determined. Not determined. Not determined. X6PNDKYEPF phage display library 2670 AEWPSLTEIKTLSHFSV AEWPSISTLSTYTHSLH AEWPSIHHMSHLLYSTY AEWPSVRTILNTDLLHP AEWPSPTRVISTTYFGS AEWPSPHKIMSTLQYLR AEWPTTRELRSLKSFLT AEWPKMTALQSTMKYVT AEWPKSFLMWMPKATQL AEWPRLSTLASMTNKAI AEWPRVKDLSTYLEGHV AEPWPMLKQLRLLKSSL AEPMNWPTVHAIRSLRK AEHISWPTLAQMSLMNF AELAHWPPVKTVLRSFT AEVTKWPNLTQLRMLAT AEGSTNRWPTVAKLMST AEPWKLIRDSLQLNYLP AEGSWLQLLNLMKQMNN AEASWTTVRSHFGSTMQ AEQLSWKALVTSVLSPT AELSELWRNYSVRLMSS AEAQLNRWKSLSKTMMS AETPSTSKWHQLIKSHR AEISLPQLMJLTHRLKQ AEFPLLPY7WMARHHGS AENLTPQYFKKWQDLTK AETSSLRNTLQMJRSLI 28 Phage display (competitive panning) 3 PMID:8226817 Calmodulin, CaM Not determined. Not determined. Not determined. X15 M13 phage display library Calmodulin was coupled to CNBr-activated Sepharose 4B (1 mg of protein/ml of resin). An aliquot (e12 pfu) of the random sequence bacteriophage library containing 1 mM CaCl2, was applied to a column of calmodulin-Sepharose. The column was then washed successively with sterile column buffers containing 1 mM CaCl2, then 1 mM CaCl2 and 1 M NaCl, and eluted with the calcium chelator, 1 mM EGTA and 1 M NaCl. The EGTA-eluted bacteriophage were amplified. An aliquot of the amplified bacteriophage was reselected two times. 2671 SSWDVLREAFTSRHPADLVHQADSQLSRASR SSWDTVRERLLKSYTADHSKTPPNRTAISSR SSRWEIVRTGLLTRPAGITNASPPTITESRR SSKWDLLRGVFWEGDRGDALSGTDTHGRTSR SSGWERVRSWAASSSAARNTSVSVTPSDQSR SSSHWDVLRGAVTLPAADSNAGRSWRTSTSR SSSRDHWSMLRGCFSSAGCSYWPDSRSHINSR SSSYALRWDALRDCIAAGCHRTDHYVRSVDSR SRCEADLHWALDRWSAAVKAGGTMPGSACSR SSGDARGSHWGFLRSAVNSSQLINTRSLTSR SSSTSNRTPGWERLRAAVNNGMKSLNDLGPSR SSATGGSTASRWAAARLRSFSPPSVIQSR 12 Phage display (common panning) 3 PMID:8635738 Calmodulin, CaM Not determined. Not determined. Not determined. R26 M13 phage display library (SRX12(A/S/P/T)A(A/D/E/G/V)X12SR) CaM (0.2 μg/well) was immobilized to 96-well polystyrene microtiter plates by non-specific absorption in 0.1 M NaHCO3 (pH 8.5). Twelve CaM-binding peptides contained the sequence motif: +W-OλR where +, -, O and λ are positively charged, negatively charged, hydrophobic, and Leu/Val, respectively. 2672 SSTSSVGRAFDVWRAAVLYSTHAVPPEQSSR SSLGLNSGDRTGWRAAVDQLLRLNKNKFDSR SSAVTDPATRSTKWAAAVADIIRSKNMQKSR WRAAV 3 Phage display (common panning) 3 PMID:8635738 Calmodulin, CaM Not determined. Not determined. Not determined. R26 M13 phage display library (SRX12(A/S/P/T)A(A/D/E/G/V)X12SR) CaM (0.2 μg/well) was immobilized to 96-well polystyrene microtiter plates by non-specific absorption in 0.1 M NaHCO3 (pH 8.5). Biotinylated CaM (0.2 μg/well) was immobilized using streptavidin (0.5 μg/well) absorbed as above. For libraries screening, e11 phage particles were incubated with immobilized CaM, non-binding phage washed away, binding phage recovered by acid elution and amplified in E. coll. Three rounds of screening were performed. 2673 SSSELSSSSRWRGLAAALMGYDTSSQQGPSR SSERDSIGRMWRQSAAALRSSFAHSQTNDSR W-R-x-x-A-A-A-L 2 Phage display (common panning) 3 PMID:8635738 Calmodulin, CaM Not determined. Not determined. Not determined. R26 M13 phage display library (SRX12(A/S/P/T)A(A/D/E/G/V)X12SR) CaM (0.2 μg/well) was immobilized to 96-well polystyrene microtiter plates by non-specific absorption in 0.1 M NaHCO3 (pH 8.5). Biotinylated CaM (0.2 μg/well) was immobilized using streptavidin (0.5 μg/well) absorbed as above. For libraries screening, e11 phage particles were incubated with immobilized CaM, non-binding phage washed away, binding phage recovered by acid elution and amplified in E. coll. Three rounds of screening were performed. 2674 SSADHALGEARRANTADKSSWPSVKRVLHSR SSTPNSELGVYKQYSAANIFRSWASRAASR SRGNGERELWWKAFSAVTEGKIKKAPGHTSR 3 Phage display (common panning) 3 PMID:8635738 Calmodulin, CaM Not determined. Not determined. Not determined. R26 M13 phage display library (SRX12(A/S/P/T)A(A/D/E/G/V)X12SR) CaM (0.2 μg/well) was immobilized to 96-well polystyrene microtiter plates by non-specific absorption in 0.1 M NaHCO3 (pH 8.5). Biotinylated CaM (0.2 μg/well) was immobilized using streptavidin (0.5 μg/well) absorbed as above. For libraries screening, e11 phage particles were incubated with immobilized CaM, non-binding phage washed away, binding phage recovered by acid elution and amplified in E. coll. Three rounds of screening were performed. 2675 CLRSGRGC(12) CLRSGFGC(5) 2 Phage display (subtractive panning) 3 PMID:9261123 Echovirus 22, EV22 Not determined. Not determined. Not determined. CX5C+CX6C+CX7C+CX9 phage display library pool Prior to panning with the virus, the phages were incubated for 1 h at 4°C in BSA-coated wells in 500 ml of Tris-buffered saline supplemented with 1 mg/ml BSA and 0.5 mM MgCl2. 2676 CVWDWGDC(11) CVWDLGRC(2) CVWDQGIC(2) 3 Phage display (subtractive panning) 3 PMID:9261123 Coxsackievirus A9, CAV9 Not determined. Not determined. Not determined. CX5C+CX6C+CX7C+CX9 phage display library pool Prior to panning with the virus, the phages were incubated for 1 h at 4°C in BSA-coated wells in 500 ml of Tris-buffered saline supplemented with 1 mg/ml BSA and 0.5 mM MgCl2. 2677 (D/N)TPVCYMNWCVE(S/T)D N(N/T)KRCYMDLCIQTP ANTWCYVDECMRIA ETYGCFMDWCKLVT 4 Phage display (common panning) 3 PMID:11298226 Anti-aflatoxins B1 and G1 monoclonal antibody 24, MAb 24 Not determined. Not determined. Not determined. f88-Cys4 phage display library (X4CX4CX5) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2678 (E/M)GTICP(M/T)DIKGCN(H/Q)TP GCEQCYVDYCYCSDAG GCEQCYVDYCYCSDAG MRGQCYMNQNMCKHPA 4 Phage display (common panning) 3 PMID:11298226 Anti-aflatoxins B1 and G1 monoclonal antibody 24, MAb 24 Not determined. Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2679 FHPRCNEMTCHIKP NNPTCPWLTCPLPS NTNHCYMDHCIQTH VANGCEKPWCNTTR TRW(N/D)C(P/R)TTYCPPSG 5 Phage display (common panning) 3 PMID:11298226 Anti-aflatoxins B1, G1 and B2 monoclonal antibody, MAb 13 Not determined. Not determined. Not determined. f88-Cys4 phage display library (X4CX4CX5) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2680 RQSYCHPWEAICHQHK RQSYCHPWEAICHQHK VDLWCPPAPWQCLPSD 3 Phage display (common panning) 3 PMID:11298226 Anti-aflatoxins B1, G1 and B2 monoclonal antibody, MAb 13 Not determined. Not determined. Not determined. f88-Cys6 phage display library (X4CX6CX4) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2681 AIGTAFL(6) INNALPT(4) LPNNALP(1) NNNALPT(1) NPDVTRS(1) NPDDRKA(1) LRTTAKL(1) LLTTAKP(1) MRTTSKT(1) LSLVPPA(1) LLTTNKD(1) 11 Phage display (subtractive panning) 3 PMID:17236253 MH01 plasma Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2682 TNCVGKDRCVTL(9) HACTGKLRCTTT(6) HKYPCSTTNCSP(6) NWGYSGMLAKIA(5) SHFPPWSLAWTH(4) MCQGKNICTTIQ(3) EQVQSPPWAPAW(1) HSVKLPPWSSVW(1) YPPWNLTWLSTP(1) KTAIGQLSSTLL(1) KAPLSTLSGSLL(1) VTPTRILSSSFT(1) KGFQPSLPLWPR(1) EGPSPLNNATLT(1) TLNSNTTKPPLA(1) GSLIQHTQVPWE(1) KLVMHTAVPYHI(1) TCITKTADLTRR(1) TDITKTADLTRR(1) QNNALPYPVSPL(1) TATPDLTLYMPG(1) 21 Phage display (subtractive panning) 3 PMID:17236253 MH01 plasma Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2683 CLGPGRAFC(18) CNLNTSKEC(3) CNNNTSKAC(1) CNNNASPDC(1) CNPDDRKAC(1) 5 Phage display (subtractive panning) 3 PMID:17236253 MH01 plasma Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2684 VPWETVW(4) VPWNAAW(2) LRTTAKL(1) LLTTAKP(1) MRTTSKT(1) LSLVPPA(1) LLTTNKD(1) AVPIWAR(1) 8 Phage display (subtractive panning) 3 PMID:17236253 MH04 plasma Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2685 TNCVGKDRCVTL(9) HACTGKLRCTTT(6) HKYPCSTTNCSP(6) SHFPPWSLAWTH(4) EQVQSPPWAPAW(1) HSVKLPPWSSVW(1) YPPWNLTWLSTP(1) KTAIGQLSSTLL(1) KAPLSTLSGSLL(1) VTPTRILSSSFT(1) KGFQPSLPLWPR(1) AKTITTGPLSSF(1) TLNSNTTKPPLA(1) ELPPWSSVWRTP(1) YKQPIDGGRLLF(1) TATPDLTLYMPG(1) 16 Phage display (subtractive panning) 3 PMID:17236253 MH04 plasma Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2686 CLGPGRAFC(18) CAWAPPWEPAWC(8) CNTNTGKTC(4) CNLNTSKEC(3) CNNNTSKAC(1) 5 Phage display (subtractive panning) 3 PMID:17236253 MH04 plasma Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2687 SKTIMAGPELRL(1) TLNSNTTKPPLA(1) LSPQFLSPTHWP(1) TATPDLTLYMPG(1) 4 Phage display (subtractive panning) 3 PMID:17236253 MH03 plasma Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2688 SVLSVWEIPGKL(2) ASAKWSIGPGRA(1) GIIWDHSSLPTL(1) 3 Phage display (subtractive panning) 3 PMID:17236253 MH06 plasma Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2689 KSLHYGPYRTSL(3) RLIEKTTTPWGP(1) 2 Phage display (subtractive panning) 3 PMID:17236253 MH07 plasma Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2690 LLSTPWY(1) 1 Phage display (subtractive panning) 3 PMID:17236253 MH05 plasma Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2691 TLNSNTTKPPLA(1) TATPDLTLYMPG(1) 2 Phage display (subtractive panning) 3 PMID:17236253 MH05 plasma Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2692 CDSPCVACC(1) 1 Phage display (subtractive panning) 3 PMID:17236253 MH05 plasma Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2693 STSPTHR(1) 1 Phage display (subtractive panning) 3 PMID:17236253 MH02 plasma Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2694 FCAGALTCTTLP(8) TLNSNTTKPPLA(1) TATPDLTLYMPG(1) 3 Phage display (subtractive panning) 3 PMID:17236253 MH02 plasma Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2695 CACSGRLTC(8) 1 Phage display (subtractive panning) 3 PMID:17236253 MH02 plasma Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Paramagnetic beads were precoated with a goat anti-human Fcλ-specific antibody and incubated (2 h at room tmeperature) with patient plasma (1:100). Beads were washed six times with PBS/0.25% gelatine/0.1% Tween-20 and rotated overnight at 4°C with 10 μL (1.5e11–2e11 particles) of the original phage peptide libraries. The next day, beads were washed extensively with PBS/0.25% gelatine/0.1% Tween-20 and the bound phages were eluted by pH shift [0.2 Mglycine-HCl (pH 2.2)/1 mg/mL BSA], neutralized with 1 M Tris-HCl (pH 9.1) and used for a negative selection with an HIV-negative plasma pool as described above. Supernatants of the negative selection were amplified and used for a second and third round of selection. 2696 LTFEHYWAQLTS(4) QQMHLMSYAPGP(1) TIRPSTTMDSPT(1) YANPQMEKAFES(1) LPNLTWALMPGA(1) YANPQMEKAFAS(1) LLADTTHHRPWT(1) 7 Phage display (common panning) 4 PMID:17875722 p53-binding domains of MDM2 Cellular tumor antigen p53 1YCR,4HFZ, Not determined. Ph.D.-12 phage display library (X12) 2697 LTFEHYWAQLTS(7) FAPLNRTVETSP(1) YAVSSSPRVAAL(1) VVHVPNSATPPR(1) 4 Phage display (common panning) 4 PMID:17875722 p53-binding domains of MDMX Cellular tumor antigen p53 3DAB, Not determined. Ph.D.-12 phage display library (X12) 2698 LCSPLV(4) LVYMVL(3) 2 Phage display (common panning) 5 PMID:18542831 PDZ domain of rho guanine nucleotide exchange factor 11 Not determined. Not determined. Not determined. X6 M13 phage display library The panning experiment was carried out with the library packed in the absence of IPTG. 2699 FAPFAL(1) VYGWLV(1) VARVYW(1) MRSVHV(1) LWSFSK(1) LLTYRP(1) HQESRM(1) LRSSPW(1) GPVILA(1) MEARLD(1) YVCRFA(1) YALRRA(1) LSVLDA(1) SVVLPR(1) 14 Phage display (common panning) 5 PMID:18542831 PDZ domain of rho guanine nucleotide exchange factor 11 Not determined. Not determined. Not determined. X6 M13 phage display library The panning experiment was carried out with the library packed in the presence of IPTG, which increased the number of fusion P8 protein copies on each phage particle as a consequence of Lac induction. 2700 LFFMRL(1) VQSSQI(1) TISLAL(1) RFGLRS(1) GFTHQS(1) NGCKST(1) GASTVS(1) YYSWSV(1) YGGGSF(1) GSGSHF(1) APPQYH(1) MDPTRP(1) CPRSLQ(1) 13 Phage display (common panning) 4 PMID:18542831 PDZ domain of rho guanine nucleotide exchange factor 12 Not determined. Not determined. Not determined. X6 M13 phage display library The panning experiment was carried out with the library packed in the presence of IPTG, which increased the number of fusion P8 protein copies on each phage particle as a consequence of Lac induction. 2701 LLHLRL[2.89] LALFLA[3.74] LRQVWL[3.5] PLALLH[3.14] QLGLNP[3.89] TNFMLP[3.69] TWLGQG[3.56] SPLTSM[2.53] VSDVVW[3.73] 9 Phage display (common panning) 4 PMID:21666888 PDZ domain of segment polarity protein dishevelled homolog DVL-2 Not determined. Not determined. Not determined. X6 M13 phage display library ELISA ELISA signal/control ratio is shown. The phages were propagated in the absence of IPTG to obtain medium peptide display level and allow the selection of a wider range of interacting sequences. 2702 CWGHSRDEC(15) CSGGRRLEC(4) G-[HR]-[RS]-R-[DL]-E 2 Phage display (common panning) 4 PMID:16239579 Aspergillus conidium Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2703 CLLSATPSC(11) ATPS 1 Phage display (common panning) 4 PMID:16239579 Aspergillus conidium Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2704 CGGVRGLDC(6) CSGGRRLEC(4) CTGGRVLSC(1) G-G-R-[GR]-L-[DE] 3 Phage display (common panning) 4 PMID:16239579 Aspergillus conidium Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2705 CGARASGSC(5) CARADAATC(2) G-A-R-A-[DS]-[AG]-[AS] 2 Phage display (common panning) 4 PMID:16239579 Aspergillus conidium Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2706 ATRDGSS(4) SGSSIDQ(1) GSSSGNF(1) GSS 3 Phage display (common panning) 4 PMID:16239579 Aspergillus conidium Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2707 DANRVGG(1) GAIRSGL(1) GWQGFGS(1) SGEVGRG(1) GGWGPGS(1) AGSGLSN(1) AGGQKSI(1) TGGRVLS(1) VGVRPDS(1) GWHPGQE(1) XXHAGQA(1) EGLRRSA(1) EYLGLAR(1) A-x-R-x-G, G-x(2)-G-x-G, A-G-x(3)-S, G-x-R-x(2)-S, H-x-G-Q, G-L-x-R 13 Phage display (common panning) 4 PMID:16239579 Aspergillus conidium Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2708 CRESVRDRC(1) CNEYSRPGC(1) CXAGPXRTC(1) CTVDSARSC(1) CDGRGMAC(1) CXTRSGHXC(1) CLEWTGLDC(1) CATGTHGPC(1) 8 Phage display (common panning) 4 PMID:16239579 Aspergillus conidium Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2709 CGPIVSFGC(1) CAGTGVSGC(1) CGGSKVSAC(1) CRNGSHVSC(1) CGAAVSILC(1) CGAVVSVDC(1) G-[AST]-x-V-S 6 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2710 CGGRLGPFC(2) CGDRLPHFC(1) CRSIGRLGC(1) CPAEGSLGC(1) [GD]-[RS]-L-G 4 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2711 CVLLRSSGC(1) CFLLRSNDC(1) CRLWRSTGC(1) CVPLSRSTC(1) L-x-R-S-x 4 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2712 RALAHPRDHPDL[1.25] ATCSMLLSRNEA[1.11] 2 Phage display (subtractive panning) 3 PMID:24779651 Esophageal cancer cell line Eca109 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Phage ELISA was used to test the specific binding of the selected phage clones to Eca109 cells. Normal human esophageal epithelial cells and bovine serum albumin cells served as controls. Absorbance values at 490 nm were subsequently recorded using a plate reader. The data were reproduced from the graph. Three rounds of subtractive screening of a phage peptide library were performed using Eca109 as the target cells and normal human esophageal epithelial cells as the absorber cells. For the second and third rounds of selection, the volume of added phage remained the same, yet the incubation time with the Eca109 cells decreased to 45 min and 30 min, respectively. In contrast, the incubation time with the normal esophageal cells increased to 1.25 h and 1.5 h, respectively. While ELISA, immunofluorescence and immunohistochemistry assays were used to validate the binding affinities of two positive phage clones, sequencing of the positive clones did not find any homology between the sequences obtained and a protein database. 2713 CKSLENSYC(18)[23.4 ± 1.1] CKSVGNYQC(2)[3.6 ± 0.8] CKNPTTGTC(2)[4.3 ± 0.3] CKSLENSQC(1)[NT] CNPSNRNDC(1)[NT] CKSPGNYQC(1)[NT] CKSLEQSYC(1)[NT] CNPSNRQDC(1)[NT] CTGTTNQYC(1)[NT] CNSPRPSTC(1)[NT] CNVQHNKTC(1)[NT] CSTRHTYDC(1)[NT] CIKSTHNHC(1)[NT] 13 Phage display (subtractive panning) 6 PMID:24841092 Bisphenol A, BPA Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Binding assay The binding specificity and relative affinity of the selected peptides to BPA were compared by assessing the number of BPA bound phages with varying peptide sequences displayed. Wild type M13 phage was used as a control. The number of wild type phages bound to BPA was 2.3 ± 0.2. NT denotes not tested. Two rounds of negative panning followed the six rounds of positive screening procedure. The collected phages from the final round of positive panning were incubated in a microcentrifuge tube filled with Binding Buffer lacking BPA for 1 h and centrifuged as above in order to exclude phage particles with cross binding affinity to the tube surface. 2714 CSMSARQLC(20/25) 1 Phage display (common panning) 4 PMID:24960578 Beta-amyloid protein 42 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Twenty-five Aβ42-binding clones with higher absorbance values were selected and amplified. Twenty clones showed the same peptide sequence: CSMSARQLC. His-tag-conjugated peptide CSMSARQLC bound to Aβ42 in a concentration-dependent manner. The peptide recognized Aβ13–35. In addition, molecular dynamics analysis and ELISA showed that the peptide predominately binds to I31 of Aβ42 by leucine and may also bind to F20 by alanine. 2715 DSRLEPNT ASRAPSST GGSVPTET DRATSSNA 4 Phage display (common panning) 3 PMID:24977927 Envelope glycoprotein E2 Not determined. Not determined. Not determined. f8-8mer phage display library (X8) Four phage clones specifically binding the E2 protein of CSFV were screened from f8/8 landscape phage library, and the phage E2P4 displaying octapeptide DRATSSNA significantly inhibited the CSFV infection in PK-15 cells at a higher titer. 2716 GVIMVIAVSCVF(11)[0.952 ± 0.244] GSLVMLVFGYMG(4)[0.947 ± 0.239] 2 Phage display (subtractive panning) 3 PMID:24979051 Sera IgG from patients with PMI Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The original phage library as a negative control. The absorbance value at 450 nm was measured. In the first round of affinity selection, microtiter wells were coated with purified sera IgG from patients without PMI to absorb nonspecific phages from the phage peptide library, and the unbounded phages were then incubated with sera IgG from patients with PMI to screen for phages containing potential peptide biomarkers. Another two rounds of affinity selection were carried out in the same way. Among the 17 positive phage clones, 11 had the same peptide sequence GVIMVIAVSCVF and four had the same peptide sequence GSLVMLVFGYMG. 2717 CSPLTENKC(7) CTALNENKC(1) CSQKSSLMC(1) CSSSSVGTC(1) CFPKKPNAC(1) CPKSTQPQC(1) CQYMDRSQC(1) CNRAFVTDC(1) CPHRGNEDC(1) 9 Phage display (common panning) 3 PMID:24689774 Anti-buffalo β-lactoglobulin antibody from rabbit 1 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2718 CGALEENKC(1) CSPYQENRC(1) CPLNENKDC(1) CPTNENREC(1) CVAGQSPKC(1) CTHAPLANC(1) CNALSENKC(1) CNPGSENKC(1) CSPLLENRC(1) CPFPRSAAC(1) CPLHENKSC(1) CGPQSENRC(1) CPYSENRAC(1) CSKTPPLTC(1) CPFDENKSC(1) CSALAENKC(1) 16 Phage display (common panning) 3 PMID:24689774 Anti-buffalo β-lactoglobulin antibody from rabbit 2 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2719 QAWTALT SHLNSWL YRAPWPP QGSTTPN TQWASYE SVTTKLN HTTSTTP TPRFLNY GSLYNLH YLRPPSA TVPGPSG TLVGDIV GMRWSEF NPAIISI VPPYWMS 15 Phage display (competitive panning) 4 PMID:24819195 β2-glycoprotein I, β2GPI Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Ultimately phage clones were obtained in elution step with polyclonal HAv anti-β2GPI antibody. 2720 ARLQLWL QPWPTST RADDLWL FQASLLP IAAHNPL AALTHIW AFSPLTI SGVAPRL RIGPELH 9 Phage display (competitive panning) 4 PMID:24819195 β2-glycoprotein I, β2GPI Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Ultimately phage clones were obtained in elution step with monoclonal chimeric IgG anti-β2GPI antibody (HCAL). 2721 HYYPWLP SHATGPW TTAPLLP MTTPSLH FVLPANY GLLASRS VSLAGIA FMMPHHQ VRHINIF WSRPATL LSWMPVV VNSSNYS 12 Phage display (competitive panning) 4 PMID:24819195 β2-glycoprotein I, β2GPI Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Ultimately phage clones were obtained in elution step with polyclonal LAv anti-β2GPI antibody. 2722 SAYHPWP MGTQLSW QLSTSAH IPKTPYS NPKIVLT LSKYPSA VTSYPFF TGSFPAL 8 Phage display (common panning) 4 PMID:24819195 β2-glycoprotein I, β2GPI Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2723 CKYMSVWKC CHSMSSHQC 2 Phage display (common panning) 4 PMID:24819195 β2-glycoprotein I, β2GPI Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2724 SLDSDRS(3/50) 1 Phage display (common panning) 3 PMID:24819195 Monoclonal chimeric IgG anti-β2GPI antibody, HCAL Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Fifty phage clones were randomly picked. Three phage clones exhibited significantly higher binding to HCAL than to background in preliminary ELISA and were identical in their primary structure. 2725 KMDGNHP(1/20) 1 Phage display (common panning) 3 PMID:24819195 Low avidity (LAv) IgG anti-β2GPI polyclonal antibody from patient B Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) From each selection experiment, 20 phage clones were randomly picked and tested for their affinity towards corresponding polyclonal anti-b2GPI. Only one phage clone with displayed peptide KMDGNHP significantly and selectively bound to polyclonal LAv anti-β2GPI fraction. 2726 FNPYWYV(11/20) QGPAHSK(2/20) 2 Phage display (common panning) 3 PMID:24819195 High avidity (HAv) IgG anti-β2GPI polyclonal antibody from patient A Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2727 FNPYWYV(6/20) 1 Phage display (common panning) 3 PMID:24819195 High avidity (HAv) IgG anti-β2GPI polyclonal antibody from patient B Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2728 CIGGRSTLC(1) CARIGSRSC(1) CGVRSSSAC(1) G-x-R-S 3 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2729 CRGFAWSPC(1) CRSERWSGC(1) CRTSGWSEC(1) R-[GST]-x(2)-W-S 3 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2730 CDMLGASVC(1) CASVHTLDC(1) CASVARIIC(1) ASV 3 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2731 CAAGIGGDC(1) CDMAAGMAC(1) CWGVAAGGC(1) AAG 3 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2732 CGDASGVVC(1) CGDGSWVGC(1) CGGQHSEVC(1) G-[DQ]-x-S-x-V 3 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2733 CLLGRISRC(1) CLLLGRFAC(1) CEIALRGRC(1) L-x-G-R 3 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2734 CDLTSNSRC(1) CALKSSSNC(1) CRPLGSKSC(1) L-x-S-[NSK]-S 3 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2735 CVGTDYVGC(1) CVLNDLVAC(1) CRMFNSVAC(1) CWMFGSGAC(1) CNPGYWGNC(1) CYYPGYDAC(1) CGALGDXXC(1) CQALGQLDC(1) CIGRAPQMC(1) CGRFMQLLC(1) CGLDTRGGC(1) CGAGHAGGC(1) CIHEYGVQC(1) CTRYEVGVC(1) CGRAFTSIC(1) CRYLRAVTC(1) CPIGLGLVC(1) CGPHKGLVC(1) CLGVSLIAC(1) CYFGVSRSC(1) CPFVAYDPC(1) CGSPVAYSC(1) CKSSTTASC(1) CSTTRENHC(1) V-x-[NT]-D-x-V, M-F-x-S-x-A, PGY, ALG, G-R-x(2)-Q, G-x(4)-G-G, E-x-G-V, R-A-[FV]-T, G-G-L-V, [LF]-G-V-S, VAY, STT 24 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2736 CRWSEKMRC(1) CRIWSDYRC(1) CLLYSNGRC(1) CLSRDRVRC(1) CRWTLAGCC(1) CTGRRHSVC(1) CXKXRELXC(1) CWRQSTEGC(1) CFHPLRDGC(1) CFDWQGRVC(1) CGQDTGSLC(1) CQPREFELC(1) CXCDSSVSC(1) CYNSLVTHC(1) 14 Phage display (common panning) 5 PMID:16239579 Aspergillus hypha Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library Screening of the phage library was performed by the BRASIL method. 2737 HWYDSFVPWGHQ(16) IPSPLEFLSELM(7) KMPIPSPAMPFG(1) SSAWWSYWPPVA(1) LPLMKTNEYPDL(1) GYFSPRISPSPS(1) 6 Phage display (common panning) 4 PMID:16700558 Phosphatidylcholine liposome Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The screen was designed to identify peptides that bind to multilamellar vesicles (MLVs) at pH 5. 2738 MHGKVHHPLSPR(5) MGHSHNTPAPKS(2) GHFNKHMRASVP(2) LTHHTSPPPVPA(1) MHHAPRLNPPVM(1) MHSHWQRPQSPF(1) MHHPVYPFQHPP(1) LTTHNTWFHHRS(1) LSHHKTVPIDAN(1) AHFGKPPHFGSS(1) MHRHLGPALSDA(1) MHSPHRTLPILT(1) YHPRALTPPTPL(1) 13 Phage display (common panning) 4 PMID:16700558 Endosomal lipid mix, ELM Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The screen was designed to identify peptides that bind to multilamellar vesicles (MLVs) at pH 5. 2739 TVPPKAPRSSDL LTRPNHGNTVDT 2 Phage display (common panning) 1 PMID:17622443 Silver power Not determined. Not determined. Not determined. Ph.D.-12 phage display and wild type library pool Forty-four binding phages were sequenced after five acid washes. Only two sequences were given out. The silver binding peptides were enriched in glycine, leucine, proline and serine compared with the frequencies of these amino acids in the library. 2740 SRLTHSNYATPT EHTNPILSHTHN QSFSTNVLHTHH 3 Phage display (common panning) 1 PMID:17622443 Platinum Not determined. Not determined. Not determined. Ph.D.-12 phage display and wild type library pool 2741 CPTSTGQAC 1 Phage display (common panning) 1 PMID:17579466 Polycrystalline platinum powder Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2742 APWHLSSQYSRT(53) IDTFYMSTMSHS(21) ALTLHPQPLDHP(12) TALATSSTYDPH(12) VTKHLNQISQSY(6) WSSGMTPDTGAP(6) ALSSSSNTTTRV(4) SSLGLTVSSIMY(3) NMNTHIHKDRPP(2) SMRLPLLSSHAL(1) VSPLSFGSPRYP(1) WSPAPHYIMGTT(1) STLPIPHEFSRE(1) 13 Phage display (common panning) 3-4 PMID:19095299 Bone-like mineral film and hydroxyapatite sintered disks Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Three to four rounds of panning were performed for each substrate. The entire experiment was repeated 3 times. 2743 KLPGWSG(2) AFILPTG(2) KPWYNDN(1) KPRQKDN(1) KPVQLDH(1) DPGFSPR(1) QDPGRAG(1) KLLGLSG(1) EFAFTVP(1) LSYKTSP(1) QQISTQM(1) NNTMMHH(1) 12 Phage display (common panning) 3 PMID:22725630 82 nm silica nanoparticles Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2744 IPTLPSS(1) NAIAPHR(1) HKYISAT(1) APSTPTP(1) HAIYPRH(1) SVASSLS(1) DRMPQYF(1) AFILPTG(1) HPFEHFS(1) KLPGWSG(1) 10 Phage display (common panning) 4 PMID:22725630 82 nm silica nanoparticles Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2745 KLPGWSG(10) 1 Phage display (common panning) 5 PMID:22725630 82 nm silica nanoparticles Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2746 LDHSLHS(2) VDNLLNL(1) VSSSLNL(1) LDHSLHQ(1) MDHSLHS(1) MDNSLHS(1) MDHWLHL(1) MDNSLHL(1) VDHVFKC(1) 9 Phage display (common panning) 3 PMID:22725630 82 nm silica nanoparticles Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2747 CTSPHTRAC(0.6) CSYHRMATC(0.3) 2 Phage display (subtractive panning) 3 http://onlinelibrary.wiley.com/doi/10.1002/adma.200502279/abstract Germanium thin films on silicon, Ge-on-Si Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Two rounds of positive selection, followed by two rounds of negative selection, followed by a final positive selection were performed. Negative selection was performed on a complementary substrate, in this case a germanium wafer. 2748 CRAKSKVAC(5) 1 Phage display (common panning) 1 PMID:14667505 Dysplastic skin Not determined. Not determined. Not determined. CX7C T7 phage display library 2749 CRAKSKVAC(5) 1 Phage display (common panning) 2 PMID:14667505 Dysplastic skin Not determined. Not determined. Not determined. CX7C T7 phage display library Forty-eight clones were isolated from the second ex vivo round. 2750 CDTAVVEGL CGTKRKC CGKRK CDTRL 4 Phage display (in vivo) 2 PMID:14667505 Squamous cell cancer cell Not determined. Not determined. Not determined. CX7C T7 phage display library To isolate phage that home to SCCs in K14-HPV16 mice, two rounds of ex vivo and two rounds of in vivo selection were performed on tumors histologically confirmed as squamous cell carcinoma grades Ⅱ-Ⅳ. Forty-eight clones were from ex vivo round 2, 48 from in vivo round 1, and 96 from in vivo round 2. 2751 CRGDKGPDC(0.152) CRGDRGPDC(0.091) CRGDKGPEC(0.061) CRGDKTTNC(0.03) CRGDHAGDC(0.061) CRGDHGVEC(0.03) CGRGDNLPC(0.03) CGRGDNLAC(0.03) CEKRGDNLC(0.03) CEKRGDSVC(0.061) CSGRGDSLC(0.03) CGKRGDSIC(0.03) CTGRGDALC(0.03) CRGDSAC(0.03) 14 Phage display (common panning) 1 PMID:19962669 Prostate cancer bone metastasis cell Not determined. Not determined. Not determined. CX7C T7 phage display library 2752 CRGDKGPDC(0.136) CRGDKGENC(0.045) CGRGDSPDC(0.045) 3 Phage display (common panning) 2 PMID:19962669 Prostate cancer bone metastasis cell Not determined. Not determined. Not determined. CX7C T7 phage display library 2753 CRGDKGPEC(0.10) CRGDKHADC(0.05) CRGDHAANC(0.05) CRGDAGINC(0.05) CGRGDMPSC(0.05) CEKRGDSLC(0.05) 6 Phage display (common panning) 3 PMID:19962669 Prostate cancer bone metastasis cell Not determined. Not determined. Not determined. CX7C T7 phage display library 2754 LLADTTHHRPWT(1) 1 Phage display (in vivo) 4 PMID:20621144 Heart muscle of mdx mouse Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the first round, mdx mice were injected with 2e11 phage particles in PBS each via tail vein injection under anesthesia and sacrificed 2 h later. The heart was harvested. The phage particles homing to the heart muscle were harvested. For round 2 and 3 of the selection, 2e10 and 2e9 phage harvested from the heart muscle was injected into mdx mice respectively. For the last selection, heart tissue was also taken and phage within were amplified. 2755 LLADTTHHRPWT(1) 1 Phage display (in vivo) 4 PMID:20621144 Heart muscle and liver of mdx mouse Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the first round, mdx mice were injected with 2e11 phage particles in PBS each via tail vein injection under anesthesia and sacrificed 2 h later. The heart was harvested. The phage particles homing to the heart muscle were harvested. For round 2 and 3 of the selection, 2e10 and 2e9 phage harvested from the heart muscle was injected into mdx mice respectively. For the last selection, liver tissue was also taken and phage within were amplified. 2756 KPANLTSTPWVP(1) TNAPPANMPFRS(1) SEIGMVHRSHQW(1) WSPGQQRLHNST(1) FSPLHTSTYRPS(1) IVHPSTM(1) SKTFNTHPQSTP(1) GFAKSHPMSLPS(1) GFPKSHPMSLP(1) APFKVPSLPSNT(1) 10 Phage display (in vivo) 4 PMID:20621144 Quadriceps muscle of mdx mouse Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the first round, mdx mice were injected with 2e11 phage particles in PBS each via tail vein injection under anesthesia and sacrificed 2 h later. The quadriceps was harvested. The phage particles homing to the quadriceps muscle were harvested. For round 2 and 3 of the selection, 2e10 and 2e9 phage harvested from the quadriceps muscle was injected into mdx mice respectively. For the last selection, quadriceps tissue was also taken and phage within were amplified. 2757 KPANLTSTPWVP(2) 1 Phage display (in vivo) 4 PMID:20621144 Quadriceps muscle and liver of mdx mouse Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the first round, mdx mice were injected with 2e11 phage particles in PBS each via tail vein injection under anesthesia and sacrificed 2 h later. The quadriceps was harvested. The phage particles homing to the quadriceps heart were harvested. For round 2 and 3 of the selection, 2e10 and 2e9 phage harvested from the quadriceps muscle was injected into mdx mice respectively. For the last selection, liver tissue was also taken and phage within were amplified. 2758 CPLLYSWWC(9) 1 Phage display (common panning) 3 PMID:16312021 Prostate-specific antigen Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The binding of the anti-total PSA mAb 8F8G5 to PSA was found to enhance the enzymatic activity of the bound PSA. Therefore the library was screened with PSA immobilized into microtiter wells coated with anti-total PSA mAb 8F8G5. Twenty-seven different sequences were obtained, but only one clone, obtained nine times, gave a strong positive signal with PSA presented by 8G8F5. This clone (CPLLYSWWC) did not react with either PSA, PSA-ACTor 8G8F5 alone. Moreover, CPLLYSWWC reacted also strongly with PSA presented by 11E5C6 whereas its binding to PSA presented by anti-total PSA mAb 5D5A5 was moderate. 2759 QYLSPLVTQWEW(5) 1 Phage display (common panning) 3 PMID:16312021 Prostate-specific antigen Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The dodecapeptide library was screened with PSA immobilized into microplate wells coated with anti-total PSA mAb 11E5C6. After three biopannings, 36 clones were randomly isolated and their DNA was sequenced. The deduced amino acid sequences of the corresponding inserts identified 22 different sequences. When tested in ELISA, these different clones did not recognize PSA or PSA-ACT coated directly in the wells. However, one clone (QYLSPLVTQWEW) obtained recognized specifically either PSA or PSA-ACT presented by either 11E5C6 or 8G8F5 but not 5D5A5. 2760 ACGWHPQNACG(17) ACTWHPQVGCG(2) ACNWHPQFSCG(1) ACEWHPQSGCG(1) ACVWHPQVPCG(1) ACQWHPQNGCG(1) ACSWHPQAACG(1) ACGYHPQQGCG(1) ACYHPQGDYCG(1) ACSVPMQIYCG(1) ACSVPVQVFCG(1) ACSAPGGSLCG(1) ACSAPTGSLCG(1) VCPGHATNSCG(1) HPQ 14 Phage display (subtractive panning, competitive panning) 2 PMID:24702159 Streptavidin Biotin 1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,1X2R,2DYH,2NR6,1ILP,1ILQ,2XDF,3EZA,3EZB,3EZE,2XDF,3MJG,1J7V,1Y6K,3RY6,4K0V ,3QD6,1QTY, Not determined. ACX7CG phage display library The CX7C phage library cyclized with BSBBA was subjected to two consecutive selection rounds against streptavidin as follows. The population was first depleted of trans binders by adding and discarding 7 times 10 μL magnetic streptavidin beads. The remaining phage were exposed 5 min to the UV light and incubated with 20 μL magnetic streptavidin beads to capture cis binders. Binders were eluted by addition of biotin and buffer with a low pH. 2761 ACHPQGDLYCG(9) ACHPQGDGICG(4) ACHPQGDGVCG(2) ACHPQGDVDCG(1) ACHPQGDQVCG(1) ACHPQGEMNCG(1) ACHPQGPASCG(1) ACHPQNDSDCG(1) ACHPQNDKNCG(1) ACHPQNDIFCG(1) ACHPQNNRYCG(1) ACHPQFWSSCG(1) ACHPQFHPDCG(1) ACSSPLPQFCG(1) ACSSPDVQVCG(1) ACSSPDRDHCG(1) ACSFFGAPKCG(1) ACPAHPDKGCG(1) ACHPMAPANCG(1) HPQ 19 Phage display (subtractive panning, competitive panning) 2 PMID:24702159 Streptavidin Biotin 1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,1X2R,2DYH,2NR6,1ILP,1ILQ,2XDF,3EZA,3EZB,3EZE,2XDF,3MJG,1J7V,1Y6K,3RY6,4K0V ,3QD6,1QTY, Not determined. ACX7CG phage display library The phage-peptides were not cyclized with BSBBA and exposed to UV light. The population was first depleted of trans binders by adding and discarding 7 times 10 μL magnetic streptavidin beads. The remaining phage were exposed 5 min to the UV light and incubated with 20 μL magnetic streptavidin beads to capture cis binders. Binders were eluted by addition of biotin and buffer with a low pH. 2762 AWCHPQGGCLG(5) AGCHPQGPCQG(4) ATCHPQVPCRG(4) AKCHPQVPCKG(3) AGCFLVWACQG(2) AGCWGQWACQG(2) AMCHHPQNCVG(1) ASCWHPQFCGG(1) AYCVHPQFCRG(1) HPQ 9 Phage display (subtractive panning, competitive panning) 2 PMID:24702159 Streptavidin Biotin 1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,1X2R,2DYH,2NR6,1ILP,1ILQ,2XDF,3EZA,3EZB,3EZE,2XDF,3MJG,1J7V,1Y6K,3RY6,4K0V ,3QD6,1QTY, Not determined. AXCX5CXG phage display library The XCX5CX phage library cyclized with BSBBA was subjected to two consecutive selection rounds against streptavidin as follows. The population was first depleted of trans binders by adding and discarding 7 times 10 μL magnetic streptavidin beads. The remaining phage were exposed 5 min to the UV light and incubated with 20 μL magnetic streptavidin beads to capture cis binders. Binders were eluted by addition of biotin and buffer with a low pH. 2763 AGCHPQGPCQG(12) AKCHPQVPCKG(4) ATCHPQVPCRG(4) APCHPQVPCQG(1) AYCHPQVGCWG(1) 5 Phage display (subtractive panning, competitive panning) 2 PMID:24702159 Streptavidin Biotin 1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,1X2R,2DYH,2NR6,1ILP,1ILQ,2XDF,3EZA,3EZB,3EZE,2XDF,3MJG,1J7V,1Y6K,3RY6,4K0V ,3QD6,1QTY, Not determined. AXCX5CXG phage display library The phage-peptides were not cyclized with BSBBA and exposed to UV light. The population was first depleted of trans binders by adding and discarding 7 times 10 μL magnetic streptavidin beads. The remaining phage were exposed 5 min to the UV light and incubated with 20 μL magnetic streptavidin beads to capture cis binders. Binders were eluted by addition of biotin and buffer with a low pH. 2764 ASCFPSFFCNG(6) AGCWQAWTCVG(5) AGCWGQWACQG(4) ASCFPRWVCGG(3) AWCSHPQNCIG(2) AYCGHPQNCWG(1) ASCFPVLACIG(1) ASCFPMLFCVG(1) AFCMLRWTCQG(1) 9 Phage display (competitive panning) 2 PMID:24702159 Streptavidin Biotin 1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,1X2R,2DYH,2NR6,1ILP,1ILQ,2XDF,3EZA,3EZB,3EZE,2XDF,3MJG,1J7V,1Y6K,3RY6,4K0V ,3QD6,1QTY, Not determined. AXCX5CXG phage display library The negative selection for depletion of trans binders was omitted. Binders were eluted by addition of biotin and buffer with a low pH. 2765 AGCHPQGPCQG(10) ATCHPQVPCRG(5) AKCHPQVPCKG(3) ARCHPQGPCQG(1) AECHPQAPCTG(1) AACHPQAPCRG(1) AHCHPQAPCRG(1) ASCHPQVPCQG(1) AYCHPQVGCWG(1) 9 Phage display (competitive panning) 2 PMID:24702159 Streptavidin Biotin 1SWT,2F01,2GH7,2IZF,2IZG,2IZH,2IZI,2IZJ,2RTD,2RTE,2RTF,2RTG,2Y3F,1X2R,2DYH,2NR6,1ILP,1ILQ,2XDF,3EZA,3EZB,3EZE,2XDF,3MJG,1J7V,1Y6K,3RY6,4K0V ,3QD6,1QTY, Not determined. AXCX5CXG phage display library The phage-peptides were not cyclized with BSBBA and exposed to UV light. The negative selection for depletion of trans binders was omitted. Binders were eluted by addition of biotin and buffer with a low pH. 2766 MEWSLEKGYTIK(24/66) WGWSLSHGYQVK(5/66) 2 Phage display (competitive panning) 3 PMID:24859530 Granulocytic myeloid-derived suppressor cells, granulocytic MDSCs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After the third round of biopanning, predominant MDSC-binding peptides were identified by PCR analysis of 66 individual phages eluted from granulocytic MDSCs. 2767 MEWSLEKGYTIK(25/48) WGWSLSHGYQVK(10/48) 2 Phage display (competitive panning) 3 PMID:24859530 Monocytic myeloid-derived suppressor cells, monocytic MDSCs Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After the third round of biopanning, predominant MDSC-binding peptides were identified by PCR analysis of 48 individual phages eluted from monocytic MDSCs. 2768 HPWIPKR WPWQHHR WPWHHVR WPWHNHR W-P-W-x(3)-R 4 Phage display (common panning) 3 PMID:24573486 Envelope glycoprotein E2(661) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Twenty-five randomly selected phages from the third round of biopanning bound to HCV E2661. Four clones (HPWIPKR, WPWQHHR, WPWHHVR, WPWHNHR) had higher affinities for binding HCV E2661 protein compared to the other clones. 2769 SHSEFWDWGP[3.69 ± 0.11, 1.6] 1 Phage display (common panning) 4-5 PMID:25014588 Histone demethylase KDM1A Not determined. Not determined. Not determined. X7+X12+CX7C M13 phage display library pool ELISA In direct phage ELISA, absorbance values were measured at 490nm. Absorbances of the peptide were 0.28 ± 0.03, 0.06 ± 0.01 and 0.05 ± 0.01 when GST, BSA and skim milk acted as control target. Besides, competitive ELISA experiments were performed between free and surface-immobilized KDM protein for phage binding. And apparent EC50 values (μM) were determined by competition assay from triplicate analyses. 2770 AWDVIWDQLLQH[3.68 ± 0.07, 10] 1 Phage display (common panning) 4-5 PMID:25014588 Histone demethylase KDM1B Not determined. Not determined. Not determined. X7+X12+CX7C M13 phage display library pool ELISA In direct phage ELISA, absorbance values were measured at 490nm. Absorbances of the peptide were 0.12 ± 0.03, 0.08 ± 0.01 and 0.07 ± 0.02 when GST, BSA and skim milk acted as control target. Besides, competitive ELISA experiments were performed between free and surface-immobilized KDM protein for phage binding. And apparent EC50 values (μM) were determined by competition assay from triplicate analyses. 2771 GRMDWLGWRYEL[2.09 ± 0.14, 11] SHSMSNRAPSALVRI[0.56 ± 0.06, 3.4] 2 Phage display (common panning) 4-5 PMID:25014588 Histone demethylase KDM4A Not determined. Not determined. Not determined. X7+X12+CX7C M13 phage display library pool ELISA In direct phage ELISA, absorbance values were measured at 490nm. Absorbances of GRMDWLGWRYEL were 0.05 ± 0.00, 0.05 ± 0.01 and 0.05 ± 0.02, and that of SHSMSNRAPSALVRI were 0.05 ± 0.01, 0.08 ± 0.02 and 0.08 ± 0.01 when GST, BSA and skim milk acted as control target. Besides, competitive ELISA experiments were performed between free and surface-immobilized KDM protein for phage binding. And apparent EC50 values (μM) were determined by competition assay from triplicate analyses. 2772 CKWMDDGYC[0.27 ± 0.06, 56] CYTRNMNQC[1.89 ± 0.14, 1.3] 2 Phage display (common panning) 4-5 PMID:25014588 Histone demethylase KDM4C Not determined. Not determined. Not determined. X7+X12+CX7C M13 phage display library pool ELISA In direct phage ELISA, absorbance values were measured at 490nm. Absorbances of CKWMDDGYC were 0.07 ± 0.02, 0.05 ± 0.01 and 0.04 ± 0.01, while that of CYTRNMNQC were 0.12 ± 0.03, 0.07 ± 0.02 and 0.06 ± 0.02 when GST, BSA and skim milk acted as control target. Besides, competitive ELISA experiments were performed between free and surface-immobilized KDM protein for phage binding. And apparent EC50 values (μM) were determined by competition assay from triplicate analyses. 2773 MYPWTEPSYLSN(16)[3.67 ± 0.40] SPWTEPHYMAEP(3)[3.07 ± 0.91] TPWTERWYWTSP(3)[3.43 ± 0.35] P-W-T-E-x(2)-Y 3 Phage display (common panning) 4 PMID:25048232 Human brain-seeking breast carcinoma cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The bound peptide-phage clones were detected by phage cell-binding ELISA. ELISA. Wild-type M13 phage (Insertless-M13) lacking the peptide insert served as a negative control. The relative binding affinity positive-to-negative (P/N) ratios of peptide-phage clones to 231-BR cells are computated as means ± SD. M13 phage clone displaying MYPWTEPSYLSN sequence, specifically bound to 231-BR cells and the binding could be competitively abolished by the peptide MYPWTEPSYLSN. Thus, this peptide is a promising peptide binding to human brain metastatic breast cancer. 2774 CSHMEYPRC[2 ± 0.14] CESAYMNNC[1.86 ± 0.02] CTVRTSADC[1.88 ± 0.13] CNCDYVLTC[1.87 ± 0.14] CLMRTNDQC[1.75 ± 0.29] CLKADKAKC[1.46 ± 0.03] CGFQHIGNC[1.24 ± 0.05] CVHVAQGRC[1.14 ± 0.09] CLPMTKHVC[1.13 ± 0.1] CPIKDLLAC[1.14 ± 0.12] CLKLGEKWC[0.74 ± 0.15] CNLCPTYYC[0.5 ± 0.02] 12 Phage display (common panning) 3 PMID:25058570 Human breast cancer cell line MDA-MB-231 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Phage binding to MDA-MB-231 cells was detected by cellular ELISA. Phage lacking an insert was used as the negative control. The experiments were performed in triplicate, and results are mean ± SD average OD450 nm values. The OD450 nm value of negative control phage binding to MDA-MB-231 cells was 0.23 ± 0.04. Data shown were reproduced from Fig. 1 in the reference. Screening of the phage library was performed by the BRASIL method, which allows the separation of cell-bound from unbound phage in one step. Single-photon emission CT (SPECT) and near-infrared fluorescence (NIRF) imaging showed enrichment of peptide CLKADKAKC to the xenograft tumors in nude mice. It could be used for breast cancer molecular imaging, which may represent a new avenue for breast cancer diagnostics, staging and assessments of therapeutic response. 2775 CNLSSSWIC(24/47)[0.669 ± 0.007] CPSTLGASC(7/47)[0.632 ± 0.01] CVPRLSAPC(7/47)[0.6 ± 0.003] CVATLPAGC(5/47)[0.476 ± 0.007] CHPLRSAFC(3/47)[NT] CSHTSLESC(1/47)[NT] 4 Phage display (subtractive panning) 3 PMID:25078431 Myoglobin Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The binding activity of the first four selected phage clones to myoglobin coated on plates was determined by phage binding ELISA. Data shown were optical density (OD) at 490 nm and reproduced from Fig. 4 in the reference. Phage library was used as a control phage, which had the value of 0.077 ± 0.008 when binding to the myoglobin. Each biopanning round consists of negative selection or sub-traction of phages that nonspecifically binds to the Dynabeads and subsequent positive selection of phages that binds to myoglobin. In the negative selection, phage library in PBS was added to the BSA-blocked Dynabeads and incubated to subtract the phages that binds to the beads. Forty seven phage clones were randomly picked from phage clones of the third, fourth, and fifth round. Four phage clones(CNLSSSWIC, CPSTLGASC, CVPRLSAPC, CVATLPAGC) from the third round of biopanning had higher frequency. The analysis of binding affinity showed that the peptide CPSTLGASC had higher binding affinity (Kd= 57 nM) than did the CNLSSSWIC and CVPRLSAPC peptide (Kd= 125 nM and 293 nM, respectively). Cross binding activity to other proteins, such as bovine serum albumin, troponin I, and creatine kinase-MB, was minimal. The identified peptides can be used for the detection of myoglobin and may be a cost effective alternative to antibodies. 2776 HFYQITWLPNTFPAR(15)[0.52 ±0.03, 0.45 ± 0.05] LSTHTKLNKADIQMP(3)[0.81 ± 0.02, 0.63 ± 0.01] YRWPSTPSASRQATL(1)[0.53 ± 0.06, 0.41 ± 0.01] GAARHCQPASPATMM(1)[0.55 ± 0.04, 0.42 ± 0.02] 4 Phage display (competitive panning) 5 PMID:25081558 Anti-TsCa IgG T. saginata metacestode crude antigen Not determined. Not determined. X15 phage display library ELISA Affinities of phages to Anti-TsCa IgG were measured by phage competitive ELISA. Data shown were absorbances at 490 nm and reproduced from Fig. 3b. High titer serum was pre-incubated with TsCa (500 μg/mL) and assayed on phage-coated plates. High titer serum directly incubated with phage-coated plates as control and the values of A490 are the first value in square brackets. There was a little reduction in the absorbance compared with that in the assay without the competitor. During the first panning, an immunotube (Nunc) was coated overnight with 5 μg/mL of anti-TsCa IgG. In the second panning, 1 μg/mL of anti-TsCa IgG was used for coating, with 0.5 μg/mL in the subsequent pannings. Affinity-selected phagedisplayed peptides were obtained by competition assay after incubation at room temperature for 90 min with TsCa at a concentration 100 times higher than that of bovine IgG. The peptides HFYQITWLPNTFPAR and YRWPSTPSASRQATL shared similarity with highly conserved proteins from the Taeniidae family with known immunogenicity. 2777 TCIWQWPDWACK(1)[0.5 ± 0.02, 0.41 ± 0.03] FCMSTCSGLKCQ(1)[0.36 ± 0.02, 0.31 ± 0.02] 2 Phage display (competitive panning) 5 PMID:25081558 Anti-TsCa IgG T. saginata metacestode crude antigen Not determined. Not determined. X6PPX(Y/F)X6 phage display library ELISA Affinities of phages to Anti-TsCa IgG were measured by phage competitive ELISA. Data shown were absorbances at 490 nm and reproduced from Fig. 3b. High titer serum was pre-incubated with TsCa (500 μg/mL) and assayed on phage-coated plates. High titer serum directly incubated with phage-coated plates as control and the values of A490 are the first value in square brackets. There was a little reduction in the absorbance compared with that in the assay without the competitor. During the first panning, an immunotube (Nunc) was coated overnight with 5 μg/mL of anti-TsCa IgG. In the second panning, 1 μg/mL of anti-TsCa IgG was used for coating, with 0.5 μg/mL in the subsequent pannings. Affinity-selected phagedisplayed peptides were obtained by competition assay after incubation at room temperature for 90 min with TsCa at a concentration 100 times higher than that of bovine IgG. 2778 VHTSIRPRCQPRAITPR(1)[1.11 ± 0.01, 0.92 ± 0.01] MGIRALPPCQNARQRLS(1)[0.85 ± 0.02, 0.77 ± 0.01] 2 Phage display (competitive panning) 5 PMID:25081558 Anti-TsCa IgG T. saginata metacestode crude antigen Not determined. Not determined. X8CX8 f88-based phage display library ELISA Affinities of phages to Anti-TsCa IgG were measured by phage competitive ELISA. Data shown were absorbances at 490 nm and reproduced from Fig. 3b. High titer serum was pre-incubated with TsCa (500 μg/mL) and assayed on phage-coated plates. High titer serum directly incubated with phage-coated plates as control and the values of A490 are the first value in square brackets. There was a little reduction in the absorbance compared with that in the assay without the competitor. During the first panning, an immunotube (Nunc) was coated overnight with 5 μg/mL of anti-TsCa IgG. In the second panning, 1 μg/mL of anti-TsCa IgG was used for coating, with 0.5 μg/mL in the subsequent pannings. Affinity-selected phagedisplayed peptides were obtained by competition assay after incubation at room temperature for 90 min with TsCa at a concentration 100 times higher than that of bovine IgG. The phage-displayed peptide VHTSIRPRCQPRAITPR competed with TsCa for binding sites, reducing the reactivity by approximately 30%. 2779 CPFPSPTWC[2.95 ± 0.07] CFGLPSWAC[3.39 ± 0.28] CNGFPSWC[3.34 ± 0.1] CLGRPLWAC[3.17 ± 0.06] CPHYSTLLC[4.07 ± 0.1] 5 Phage display (common panning) 3 PMID:25154906 Anti-soluble CD14-fraction ST monoclonal antibody CD14 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Affinities of phages to anti-soluble CD14-fraction ST monoclonal antibody were measured by phage ELISA. ELISA index values were read at 492 nm and data shown were reproduced from Fig. 1. An irrelevant peptide fused to a bacteriophage was used as a negative control, which had the value of 0.53 ± 0.08 when binding to the monoclonal antibody. The competitive ELISA assay between M.H2 clone (CPHYSTLLC) and the recombinant sCD14 for binding to the polyclonal anti-CD14 antibody demonstrated its specificity to the target. 2780 CTPTFPPRC[2.35 ± 0.07] CLNPVSSSC[2.52 ± 0.34] CSTSSPSYC[1.62 ± 0.25] CSLASLPAC[2.34 ± 0.18] CTAQRLPSC[2.08 ± 0.13] CTPLLSPFC[2.58 ± 0.2] CSLLATAPC[2.76 ± 0.18] CDHGPLPRC[2.78 ± 0.19] CTLPVPLHC[2.99 ± 0.34] 9 Phage display (common panning) 3 PMID:25154906 Anti-CD14 polyclonal antibody CD14 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Affinities of phages to anti-CD14 polyclonal antibody were measured by phage ELISA. ELISA index values were read at 492 nm and data shown were reproduced from Fig. 1. An irrelevant peptide fused to a bacteriophage was used as a negative control, which had the value of 0.61 ± 0.12 when binding to the polyclonal antibody. 2781 LQWRRDDNVHNFGVWARYRL(2) EEHTHRWPFWGHQERWGKQS(1) EHGDGPGGKMRWWWHGGGTR(1) GAFWKNNGSTQPWNPEDSSL(1) GDATAKEMRSTQDTPQERGA(1) LSLGRGADRIIPWELRRPGG(1) MEGRSIGGRFRHTADMMVEA(1) RVSGDNQAPTQRNNQGAEWT(1) SQTLKGWRTGKLQPETLRWS(1) SSEFAGENGGSTRGHKFDGY(1) VAPLLRSESAIARSLVSYPQ(1) 11 Phage display (common panning) 3 PMID:25188559 NCI-H1299 non-small cell lung cancer cell line Not determined. Not determined. Not determined. X20 M13 phage display library Round 1 of panning is performed against H1299 nonsmall cell lung cancer cells. The output of round 1 is then split into two groups. H1299 NSCLC cells of this group remains untreated with chlorpromazine. 2782 AARPPTAPTEMHGAEMKGMT(1) EEGGEVKGSAHASTDDTTRF(1) GDMVPWAHPWEPWLGNKVEA(1) GTYASVTRSHDRSGERIGDH(1) KVQVERKEALGIQKIAVSRR(1) LAGRQGPERSTVENNLSGTK(1) NATWGKALRDYHRGVWSRVS(1) SKSFALDGTPERYSRTLVRR(1) VVSIPSTVGKGYPDSWAVRR(1) WEGSEGTVESDNLQNKKGGK(1) WGANQNRFAMAWAGATGASS(1) 11 Phage display (common panning) 3 PMID:25188559 NCI-H1299 non-small cell lung cancer cell line Not determined. Not determined. Not determined. X20 M13 phage display library Round 1 of panning is performed against H1299 nonsmall cell lung cancer cells. The output of round 1 is then split into two groups. This group is then incubated with H1299 NSCLC cells treated with chlorpromazine. 2783 ATEPRKQYATPRVFWTDAPG(10) LQWRRDDNVHNFGVWARYRL(1) FGSWPTGWKARAYNDLPPAR(1) 3 Phage display (common panning) 4 PMID:25188559 NCI-H1299 non-small cell lung cancer cell line Not determined. Not determined. Not determined. X20 M13 phage display library Round 1 of panning is performed against H1299 nonsmall cell lung cancer cells. The output of round 1 is then split into two groups. H1299 NSCLC cells of this group remains untreated with chlorpromazine. 2784 ATEPRKQYATPRVFWTDAPG(9) AAPRLGGTQIKSESTKMGSD(1) GAWEAVRDRIAEWGSWGIPS(1) TWDGNEAERSPGSTGEDAAR(1) 4 Phage display (common panning) 4 PMID:25188559 NCI-H1299 non-small cell lung cancer cell line Not determined. Not determined. Not determined. X20 M13 phage display library Round 1 of panning is performed against H1299 nonsmall cell lung cancer cells. The output of round 1 is then split into two groups. This group is then incubated with H1299 NSCLC cells treated with chlorpromazine. 2785 ATEPRKQYATPRVFWTDAPG(6) 1 Phage display (common panning) 5 PMID:25188559 NCI-H1299 non-small cell lung cancer cell line Not determined. Not determined. Not determined. X20 M13 phage display library Round 1 of panning is performed against H1299 nonsmall cell lung cancer cells. The output of round 1 is then split into two groups. H1299 NSCLC cells of this group remains untreated with chlorpromazine. 2786 ATEPRKQYATPRVFWTDAPG(6) 1 Phage display (common panning) 5 PMID:25188559 NCI-H1299 non-small cell lung cancer cell line Not determined. Not determined. Not determined. X20 M13 phage display library Round 1 of panning is performed against H1299 nonsmall cell lung cancer cells. The output of round 1 is then split into two groups. This group is then incubated with H1299 NSCLC cells treated with chlorpromazine. 2787 RTEVPVLSFTSPLTG[1.76] GDVWLFKTSTSHFAR[1.91] AREYGTRFSLIGGYR[0.21] HAAFEPRGDVRHTLL[2.00] LGRAGQSYPSFARGL[0.52] PIFPVVSSSGSSSSP[1.58] PLSHGSVVYPRSSLG[1.36] RRDTVPRSLSAPLSW[0.59] PAVASTSSLIIDGPF[2.29] HPPLASVWHVSVPL[0.83] LHDFRSPIYASLLGF[1.53] AGDGGLGRVAAGARV[1.15] RVFHLWPHPTSTLSA[0.25] APLSYNFASMPFMSG[0.73] HPGWFDSAWFRAVSR[1.24] ARDSRCGGFLGCGVT[1.36] AMVRGFSFGMSRGSD[1.94] RSLWSDFYASASRGP[6.57] SYSVVNSPWCDGTCD[1.25] SRDGLHSFCYVGCPP[0.40] GVGDADGFIPVISAV[0.59] PVFFRLSPVTEGGGV[1.40] FPSYPFIAYSLQTPV[1.23] RRLPHLMPFEGSVFL[3.55] GPHFDYRTGLGWRFG[1.35] LGKGLTGSALSLSAL[2.29] YGVTPSPRSPWATAH[3.43] VFVDGARYSTASDSL[2.83] GAGIFGPWGVFAAVP[1.21] GYRSAFVPFVARGGH[3.65] RYRVGFTPGTIAAVL[1.13] 31 Phage display (subtractive panning, in vivo) 4 PMID:25250205 Human ovarian adenocarcinoma cell line SKOV-3 Not determined. Not determined. Not determined. f5-15mer phage display library (X15) Competition experiment,Micropanning assay Micropanning assay was used to determine binding of selected phage clones to SKOV-3 and HS-832 cells. SKOV-3 or HS-832 cells were incubated with individual phage clones and eluted using 2.5% CHAPS. Phage binding was determined by titer and the SKOV-3 to HS-832 ratio was calculated and shown. In addition, competitive binding experiments indicated that the peptide RSLWSDFYASASRGP displayed a half maximal inhibitory concentration (IC50) value of 10.5 ± 1.1 μM. The library was pre-cleared from vasculature and non-tumor target binding phage according to previous methods. The pre-cleared phage library was screened against xenografted human OC SKOV-3 tumors in mice. In the last round of selection, 31 individual phage clones were identified, and that phage clone RSLWSDFYASASRGP exhibited the highest SKOV-3-to-HS832 binding ratios of 6.57, indicating preferable binding to the human OC cell line. 2788 VTVPTMPIRTAT FTVPTVPITALH ATVPTTPITLTP GFTVPHLPIVPE FIVPTIPIRGLP SPGIHVVPTRPI FPTYVPYAPIPP VIVPVTPIKPQA QPFVVPTSPVRG HAVVPVNPVKSL TALHYAPVPVGP TPIFVQPWTPTY VTVPTQEITPNL SIFVPTHSIEPS SVVPTVTITGNY SVVPFHEIISRG VSVPTVSIKPYI KAVVPVSSIMPF DSVTPTSVMAVA YLQQDPLPRKTY AYALPGPDPLPR NVDQGRDPLPRE WQNEDPLPRFSW LSPLGGWDPLPR KLDPLPRHFSAA ITPGQWRSPLHF LTPGQLNQQIWE LTPGQITFTQGQ LTPGQHLWWRSP TPGQLRAESAMT LTPGQAERWWAR VPTXPI, VTPXXI, DPLPR, LTPGQ 31 Phage display (common panning) 4 PMID:25273631 Anti-1/3NH2TTPI polyclonal antibody Triosephosphate isomerase (EC:5.3.1.1) Not determined. Not determined. Ph.D.-12 phage display library (X12) Fourty-one phage mimotope clones were isolated and grouped according to their amino acid sequence, finding the consensus VPTXPI, VPTXXI, LTPGQ, and DPLPR. 2789 ADANRHSTLRER TPLWQHLLSGRA ADANRRSTLRER TPPWVTVLLSRQ ADSRRALLAQRA TPQWQQLLSFRQ ADSWRALLAQRA TPSWATLLAQRA ASPWHQLLAERR TYRMDIMSIKTV ATSWKEMLAERQ WHDITSLRQYSF DSPPIFDATLPK TPWEEVLLSRLR DWREILGARSQV DPVWVNILTSRQ EHVLWQQLLTSR AQWQETLSERAR ENWRLTLLQRNG KCCYYDHSHALS GGLHWTEILRSR HSLRSDWPLRPG GGVHWSEILSYR ELTGWRLLLAQR GRLQQHEIFRSG KVVDLYSGWNRS KTPWQEMLASRI HVLWQHVVDLCR PPMWADMLLARS GPFLPLTSLHWR SCKQVLEHRQGM NWRETMGVRSQV SIDGRSIISSRN SDWTHVLSQRAL SLDWTELLRLRT KIYDLSLLHPST SMPQWQELLKVR QPAWQQTLINRS SPLWQDIILTRS SWMETLRTRNMS SPTYHSSTGLND SLSNYQIAGNGL SSPSWRDVLLSR TVSWQALLEMRQ STGDWREILRNR TPAWQATLLGRQ TDWRTQLHLRQG 47 Phage display (common panning) 3 PMID:25301558 Enterobactin synthase component E 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase Not determined. Not determined. Ph.D.-12 phage display library (X12) Upon panning immobilized EntE with a random peptide phage library, we recovered 47 unique EntE-binding dodecamer peptide sequences that aligned to a region of the EntA primary sequence corresponding to helix alpha 4. 2790 KVWLPPRHEHQY(2)[0.92 ± 0.03] KVFYPAAANPNQ(1)[0.41 ± 0.06] 2 Phage display (subtractive panning) 2 PMID:25311190 Vaccinia virus strain NYCBOH Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Affinities of selected phages to vaccinia virus strain NYCBOH were measured by phage ELISA. The OD450-620 values were given and data shown were reproduced from the Fig. 1 in the reference. The wild-type M13KO7 phage (WT) was used as a control, which had the value of 0.3 ± 0.01 when binding to vaccinia virus strain NYCBOH. A negative selection was performed before every biopanning round. In the negative biopanning, the library was added to BSA-coated wells. Then the supernatant was transferred to target-coated wells. After the fourth round, binding phages were eluted and used for a further negative selection against HEp-2 cell lysate. 2791 NRPDSAQFWLHH(3)[0.92 ± 0.06] KVWLPPRHEHQY(1)[0.92 ± 0.03] KVFYPAAANPNQ(1)[0.41 ± 0.06] QALLEGNAKGGN(1)[0.91 ± 0.04] TADKLLYGLFKS(1)[2 ± 0.05] DEWDALLMRIRT(1)[1.39 ± 0.12] 6 Phage display (subtractive panning) 3 PMID:25311190 Vaccinia virus strain NYCBOH Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Affinities of selected phages to vaccinia virus strain NYCBOH were measured by phage ELISA. The OD450-620 values were given and data shown were reproduced from the Fig. 1 in the reference. The wild-type M13KO7 phage (WT) was used as a control, which had the value of 0.3 ± 0.01 when binding to vaccinia virus strain NYCBOH. A negative selection was performed before every biopanning round. In the negative biopanning, the library was added to BSA-coated wells. Then the supernatant was transferred to target-coated wells. After the fourth round, binding phages were eluted and used for a further negative selection against HEp-2 cell lysate. 2792 NPTPYPMLPLRG(8)[0.52 ± 0.02] GDLASWIITSFK(8)[1.87 ± 0.12] TADKLLYGLFKS(6)[2 ± 0.05] KIFQLPQISPPM(4)[1.11 ± 0.04] TPVWSWEPPLQE(4)[1.93 ± 0.05] KVWLPPRHEHQY(3)[0.92 ± 0.03] DEWDALLMRIRT(3)[1.39 ± 0.12] QALLEGNAKGGN(2)[0.91 ± 0.04] KPTYSWDPAQLK(2)[0.98 ± 0.01] GPTFSWDHLRGQ(2)[0.69 ± 0.02] NMELHPHSLPRP(2)[0.61 ± 0.05] ANTTKHSVLAAI(2)[1.2 ± 0.04] EALNDWVNDSEY(2)[1.45 ± 0.06] APTAYNKNDWAL(2)[0.95 ± 0.03] DPWWRGNEARAA(2)[0.78 ± 0.02] 15 Phage display (subtractive panning) 4 PMID:25311190 Vaccinia virus strain NYCBOH Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Affinities of selected phages to vaccinia virus strain NYCBOH were measured by phage ELISA. The OD450-620 values were given and data shown were reproduced from the Fig. 1 in the reference. The wild-type M13KO7 phage (WT) was used as a control, which had the value of 0.3 ± 0.01 when binding to vaccinia virus strain NYCBOH. A negative selection was performed before every biopanning round. In the negative biopanning, the library was added to BSA-coated wells. Then the supernatant was transferred to target-coated wells. After the fourth round, binding phages were eluted and used for a further negative selection against HEp-2 cell lysate. 2793 CGGIALAGC(22) CGWHRWRVC(3) CLDLGVADC(2) CLMRFQRSC(2) CLHALRGRC(2) CRTSLTGPC(2) CLRMGFRSC(2) CRFGSLVGC(1) 8 Phage display (common panning) 5 PMID:25324134 Surface glycoprotein Tc85-45 Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library One hundred phage clones from the fifth round were randomly chosen and sequenced. Sequence alignment analysis revealed that 22 % of the phages encoded the peptide motif CGGIALAGC. 2794 CLPFRWTRC(7) CRWPALLTC(4) CWMMGPYFC(4) CRLDSFNRC(3) CGWHRWRVC(3) CWGDVPSGC(3) CSGFRVEAC(2) CRFGSLVGC(1) 8 Phage display (common panning) 5 PMID:25324134 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. fUSE5-based CX7C phage display library 2795 LLSSKTL LSFPFPG FTSFSPY QATHFHS LASLPFR THVFSWI ACDPSPN TPSLHRS TAMARSA VALLPHH QSPPALL FSLLGSL VLLGPFP YPFSLLH SLGPQIK MSPTYLL DRAALSL ALTPQLL QTSPPLA FPLFGLS 20 Phage display (subtractive panning) 3 PMID:25333662 Sera IgG from asymptomatic CVL and symptomatic CVL Leishmania infantum Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage library first incubated with IgGs from healthy dogs. The supernatant containing the clones that were not adhered to the IgGs was recovered and transferred to a new tube, and this procedure was repeated for three times. Then, the supernatant incubated with IgGs purified from asymptomatic CVL. The bound phage clones were transferred to a new tube containing the IgGs purified from asymptomatic CVL, and the process was repeated for three times. After this, the recovered phage clones were transferred to a new tube containing the IgGs that had been purified from symptomatic CVL, and the process was also repeated for three times. 2796 TGFFSQR[1.60] VFGFFNTR[1.63] TGFFATP[1.66] MPGFFEMR[1.65] NGYFARN[1.63] SGYFSNN[1.69] VGYFFER[1.63] GFFATQ[1.70] SGFFSDR[1.56] SGYFAER[1.84] SGFFESK[1.76] TGFFAKK 11 Phage display (common panning) 3 PMID:25468796 Anti-GapC monoclonal antibody 1E11 Glyceraldehyde-3-phosphate dehydrogenase (EC:1.2.1.12) Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of positive phage clones to mAb1E11 were measured by sandwich ELISA. The OD values at 405 nm were read and data shown were reproduced from the Fig. 1E in the reference. The wild-type M13 phage was used as a control, which had the value of 0.35 when binding to the mAb1E11. Eleven positive clones recognized by mAb1E11 were identified, most of which matched the consensus motif TGFFAKK. Sequence of the motif exactly matched amino acids 97–103 of the S. dysgalactiae GapC. 2797 DHIHWITPSHPG[2.15 ± 0.09] DHYSYTWFSWPT[1.55 ± 0.07] ACSYHTTRAFVC[1.52 ± 0.12] ACLYHTTRAFVC[1.73 ± 0.08] GHFKWVPYDSLY[2.05 ± 0.1] THWNWLNPYMAV[0.91 ± 0.15] ALKIWPNPPRSN[1.13 ± 0.2] WHLEWITPMASD[2.16 ± 0.05] THISWMSPQKLW[1.46 ± 0.02] THERLYWYSPSE[1.33 ± 0.2] DSLRQLPLPVLS[1.44 ± 0.27] EHMQWMRATDLF[1.73 ± 0.09] QLEWSYWPQLSR[2.03 ± 0.06] 13 Phage display (common panning) 3 PMID:25446106 3-hydroxy-3-methylglutaryl-coenzyme A reductase ( (EC:1.1.1.34)) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Binding capabilities of the selected phages toward recombinant hHMGR was estimated by ELISA. The OD values at 655 nm were measured and data shown were reproduced from the Fig. 1 in the reference. The experiments were performed in triplicate and presented as mean ± standard deviation. The original library was served as the control, which had the value of 0.12 ± 0.02 when binding to the hHMGR protein. These peptides are all hydrophilic in nature (negative values of xLogP), and most are negatively charged with approximately 30% hydrophobic residues. The results indicate that the tetrapeptide PMAS inhibits hHMGR effectively (IC50 = 68 μM), could be a lead compound to develop hypocholesterolemic agents. 2798 CTPAFRYSC(20/151) CSSHSLLYC(9/151) CGQHIERGC(9/151) CAVLTHPAC(8/151) 4 Phage display (common panning) 3 PMID:25393207 Bone marrow-derived dendritic cell Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) After the third round of biopanning, 151 phages were randomly isolated and analyzed for peptide sequence. Four phage isolates frequently appeared up to 30% of phages. Besides, CTPAFRYSC (TP) conjugated to chitosan in order to develop an efficient DC-targeting vaccine delivery carrier. 2799 TDHTHNKGYANK TSHPSYYLTGSN SHQALQEMKLPM SMESLSKTHHYR KLHISKDHIYPT NRPDSAQFWLHH DPQNHNWTNKPA YLPHMLVHGSRH TYPVVGHQQNVM DIMPKLRDDVHN NAHTSNNVVAFP YGTSMTQSNWRH SYGSLQTRFGHI KFFNNTEATTRP NYALRDPVGQRY LPSVTEILGSNF TSAVTLTSDPTL QNFSQMMSIPRK 18 Phage display (common panning) 2 PMID:25343575 9-fluorenylmethoxycarbonyl-threonine and leucine-methyl ester (Fmoc-T/L-OMe) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The biopanning process revealed 18 phage clones which are able to catalyze formation of the self-assembling reaction product in the second round of panning. 2800 AMHSLVGPAFNR HDTSEQLLVAPS DLRSCTACAVNA ATTWTVAHGVSR STDDDHLLAATT HPTGSKSTTSTY HTDSDPLLAAPS PTSEVYLFSGNF 8 Phage display (common panning) 2 PMID:25343575 9-fluorenylmethoxycarbonyl-threonine and leucine-amide (Fmoc-T/L-NH2) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The biopanning process revealed 8 phage clones which are able to catalyze formation of the self-assembling reaction product in the second round of panning. 2801 TPSYDTYAAELR(16) SVSVGMKPSPRP(4) KSIALKNTNPHA(1) SPCVQCSSGLCP(1) PCLKMGIHTTKR(1) YGLHRQAACPLT(1) 6 Phage display (in vivo) 3 PMID:25408466 Cerebrospinal fluid Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the first round, the rats (n = 3) were injected intravenously (iv) with e12 pfu of the library suspended in 100 μl TBS. The phage was allowed to circulate for some time in vivo. The rats were then anesthetized using 5 % chloral hydrate (0.4 g/kg), and phage-containing supernatant was recovered from the cerebrospinal fluid, amplified by infecting competent bacteria (Escherichia coli ER2738), titered and pooled for the next round of screening. Subsequent screening rounds were conducted by intravenously injecting the newly amplified phage (1 × e12 pfu in 100 μl TBS) and repeating the procedures described above. After labeling with FITC, TPSYDTYAAELR, referred to as the TPS peptide, demonstrated significantly greater brain accumulation efficiency. The TPS peptide represents a previously unreported promising motif that can be used to design a drug delivery system that can cross the blood–cerebrospinalfluid barrier (BCSFB). 2802 SWPLYSDASGLG(11)[0.72 ± 0.17] VSLHDDASGTHR(2)[0.92/0.85] WLGESMISGWLY(1)[0.58] TKFTDAGDTSFN(1)[0.75] EHNDFPMYTWRP(1)[0.92] WPDIEMDLGPLYA(1)[0.85] 6 Phage display (common panning) 3 PMID:17658545 Anti-Lpp20 monoclonal antibody L001 LPP20 lipoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of positive phage clones to the mAb L001 were measured by ELISA. The absorbance at 492 nm was measured and data shown were reproduced from the Fig. 5 in the reference. The phage library was served as the control, which had the value of 0.32 when binding to the mAb L001. In ELISA experiment, the peptide SWPLYSDASGLG appeared 11 times and the affinity value was presented as mean ± standard deviation, while VSLHDDASGTHR appeared 2 times and the two corresponding affinity values were separated by a slash. One mimotope (SWPLYSDASGLG) showed a good match with the Lpp20 proteins at 114–117aa (DASG) and the serum of mice induced by the phage clone clearly recognized Lpp20 protein. This mimotope provided an alternative approach for the diagnosis and development of a vaccine for H. pylori. 2803 QHYNIVNTQSRV[1.8 ± 0.08] 1 Phage display (common panning) 5 PMID:25070131 Anti-EGFR monoclonal antibody ICR-62 Epidermal growth factor receptor Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of mimotope conjugated with BSA (QHYNIVNTQSRV-BSA) to the mAb ICR-62 were measured by ELISA. The OD values at 450 nm were measured and data shown were reproduced from the Figure 2 in the reference. BSA was served as the control, which had the value of 0.26 ± 0.02 when binding to the mAb ICR-62. The affinity values were presented as mean ± standard deviation. Sequence analysis of eight reactive phages demonstrated that the interacting sequence QHYNIVNTQSRV plays a significant role in binding to anti-EGFR monoclonal antibody. 2804 GFPWHHHLVHAH[3.63] VRSHFRGHWVRF[2.81] SCYYPQLTRHRF[2.95] SSYYPQWPTDRF[4.27] SSYYPQLTAHRF[3.95] 5 Phage display (common panning) 4 PMID:25100324 Tumor necrosis factor Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbances were measured at 405 nm. The value shown is the binding fold for each phage clone, that is the ratio of its specific signal and the one generated by wild-type M13 phage. The real targrt is hexahistidine-tagged recombinant human TNFα (6His-rhTNFα). G6 peptide with the sequence of SSYYPQWPTDRF was screened and its binding to rhTNFα was investigated by isothermal titration calorimetry (ITC) experiments and molecular docking (MD) simulations. After pegylation, G6 peptide was conjugated on magnetic MPs and the system was assessed in the detection of rhTNFa in human serum. 2805 ETCRASCINESA(13/20)[1.22 ± 0.15] 1 Phage display (subtractive panning) 3 PMID:25324041 IgG from patients with CHB with HBsAg seroclearance following peg-IFN-α therapy Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of positive phage clones to sera from chronic hepatitis B patients,with hepatitis B surface antigen (HBsAg) clearance, following pegylated interferon-α therapy were measured by phage ELISA. Absorbances were measured at 450 nm and data shown were reproduced from the Figure 1A in the reference. The original phage library was used as negative control, which had the value of 0.27 when binding to the sera. In ELISA experiment, the peptide ETCRASCINESA appeared 13 times and the affinity value was presented as mean ± standard deviation. To increase the screening specificity, microtiter wells were coated with purified sera IgG from the patients with CHB without HBsAg seroclearance, in order to absorb non-specific phages from the random phage peptide library. The unbound phages were then incubated with the sera IgG from patients with CHB, with HBsAg seroclearance, to screen for phage containing potential peptide biomarkers. In the validation phase, phage-ELISA results showed that the positive reaction rate of the ETCRASCINESA (named IFNC1) peptide phage clone was 92.0% with the HBsAg seroclearance group (n=50), which was significantly higher, as compared with the randomly selected HBsAg non-clearance group (12.0%, n=50) and the healthy control group (8.0%, n=50). The newly identified mimic peptide IFNC1 showed a high predictive validity HBsAg seroclearance in patients with CHB, following peg-IFN-α therapy. Therefore IFNC1 may be a potential serum biomarker, which could be used to predict the treatment outcomes of peg-IFN-α therapy. 2806 EWLFEFPTPVDA(12) DWIATWPDAVRS(12) 2 Phage display (subtractive panning) 2 PMID:25410289 Embryonic progenitor cell line SM30 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The library was subjected to cycles of selection on SM30 cells. To subtract nonspecific binding phages from the library, the phage library was pre-adsorbed against adult dermal fibroblasts cells before each of selection cycle. 2807 LTPPGRLSSWPL(2/17) SLSLVAPVLSLL(2/17) LTARQLVTNTTH(2/17) LTLPLNTKTVQD(1/17) QPPSINLTLYPM(1/17) LTAPLNVTMNPV(1/17) SVTLSLRLPFPS(1/17) MSNQQMRALSLL(1/17) PQSLQLPRFDMR(1/17) GSFLQLLPQPSP(1/17) FHPIKLAPYPAV(1/17) TYLNYSMGIDLE(1/17) STWPAFRLFTNI(1/17) HGLIEGVSNLVF(1/17) 14 Phage display (common panning) 5 PMID:25661733 Ovarian cancer cell lines OC-3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) To test the ability of these 14 phage clones to bind to other clear cell ovarian adenocarcinoma cells, the binding activity of each individual phage clone to two other clear cell ovarian cancer cell lines(ES2, SKOV-3) was further analyzed by flow cytometry. Overall, two phage clones, LTPPGRLSSWPL and SVTLSLRLPFPS, exhibited prominent binding to the ES2 and SKOV-3 cell lines. 2808 ALRINTDPDFTE(15/19) DARMWGHMTTML(1/19) VWAMENSIRMTL(1/19) KLFDPWLVQSHF(1/19) MHPNAGHGSLMR(1/19) 5 Phage display (common panning) 3 PMID:25728640 Polyclonal rabbit anti-AK IgG Arginine kinase (AK) Not determined. Not determined. Ph.D.-12 phage display library (X12) In the second and third round of biopanning, 50 ug/mL and 25 ug/mL IgG were incubated with amplified phages selected in the previous biopanning round, to enrich those phage displaying high-affinity binding. 2809 SHSLLHH(2) SIPSLNM(1) SQTQLPG(1) SQPDRNE(1) SSLFYKY(1) SPPGSPH(1) YPHLPEH(1) LSVLGDQ(1) LIRPSPD(1) DSSPLSL(1) ASAPLLG(1) QTSPPHI(1) GTPTPAI(1) NAPYGLR(1) EHFQPPL(1) TNDTSPS(1) VIKSWYQ(1) VESTGIW(1) VLHPSRN(1) 19 Phage display (subtractive panning) 1 PMID:25754556 Cyclin-dependent kinase inhibitor 2A, isoforms 1/2/3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Prior to p16 selection, non-specific binding phages were removed from phage display library by adsorption of 2ml phage library on blank plates three times. 2810 SHSLLHH(11) YAWDTYR(3) SLHQPHL(1) SHGNWWR(1) YRAPWPP(1) FPPSVIR(1) HAIYPRH(1) IPTHIRP(1) 8 Phage display (subtractive panning) 2 PMID:25754556 Cyclin-dependent kinase inhibitor 2A, isoforms 1/2/3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Prior to p16 selection, non-specific binding phages were removed from phage display library by adsorption of 2ml phage library on blank plates three times. 2811 SHSLLHH(13)[0.54] SLHQPHL(2)[0.57] YRAPWPP(2)[0.35] YAWDTYR(1)[0.36] FPPSVIR(1)[0.57] FPPLVLR(1)[NT] 6 Phage display (subtractive panning) 3 PMID:25754556 Cyclin-dependent kinase inhibitor 2A, isoforms 1/2/3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Optical densities (ODs) of the enzymatic reaction were measured by ELISA plate reader with 405 nm filter set. ODs of the reaction in p16-coated wells were subtracted with the non-specific background signals to obtain the actual p16 binding signal of each phage clone. When the amount of phage display particles (PFU) was 6.25e10, the OD values were recorded. Data were reproduced from the graph. The OD value of wild-type phage is 0.13. Prior to selection, non-specific binding phages were removed from phage display library by adsorption of 2ml phage library on blank plates three times. 2812 GGSQGAY(2.1%) 1 Phage display (in vivo) 1 PMID:25762070 Soft tissue of prostate cancer xenograft Not determined. Not determined. Not determined. CX7C fUSE5 phage display library For round 1, tumor-bearing mice received 2e9 transducing units (TU) of phage peptide library i.v. After 24 h of circulation, the mice were perfused with 10 mM PBS (pH 7.4), and tumors were collected. Soft tissue and bonelike matrix compartments of the tumor were separated physically, weighed, and homogenized for phage recovery. The recovered phage pool was amplified and subjected to another round of selection. 2813 RIDAGTT(8.4%) PKRGFQD(4.2%) SPSQRQY(2.1%) GQVGIWS(2.1%) SGPTRGM(2.1%) GSQQQGR(2.1%) PGDQPRG(1.1%) 7 Phage display (in vivo) 2 PMID:25762070 Soft tissue of prostate cancer xenograft Not determined. Not determined. Not determined. CX7C fUSE5 phage display library For round 1, tumor-bearing mice received 2e9 transducing units (TU) of phage peptide library i.v. After 24 h of circulation, the mice were perfused with 10 mM PBS (pH 7.4), and tumors were collected. Soft tissue and bonelike matrix compartments of the tumor were separated physically, weighed, and homogenized for phage recovery. The recovered phage pool was amplified and subjected to another round of selection. The two peptides, PKRGFQD and SNTRVAP, targeted the cell surface of androgen-independent prostate cancer cells in vitro, and homed to androgen receptor-null prostate cancer in vivo. Purification of tumor homogenates by affinity chromatography on these peptides and subsequent mass spectrometry revealed a receptor for the peptide PKRGFQD, α-2-macroglobulin, and for SNTRVAP, 78-kDa glucose-regulated protein (GRP78). 2814 RIDAGTT(37.6%) SGPTRGM(33.3%) PKRGFQD(18.3%) PGDQPRG(3.2%) LDGPRAS(2.2%) 5 Phage display (in vivo) 3 PMID:25762070 Soft tissue of prostate cancer xenograft Not determined. Not determined. Not determined. CX7C fUSE5 phage display library For round 1, tumor-bearing mice received 2e9 transducing units (TU) of phage peptide library i.v. After 24 h of circulation, the mice were perfused with 10 mM PBS (pH 7.4), and tumors were collected. Soft tissue and bonelike matrix compartments of the tumor were separated physically, weighed, and homogenized for phage recovery. The recovered phage pool was amplified and subjected to another round of selection. 2815 SNTRVAP(33.7%) RLGLAWG(14.5%) SNNFVAP(3.6%) 3 Phage display (in vivo) 2 PMID:25762070 Bonelike matrix tissues of prostate cancer xenograft Not determined. Not determined. Not determined. CX7C fUSE5 phage display library For round 1, tumor-bearing mice received 2e9 transducing units (TU) of phage peptide library i.v. After 24 h of circulation, the mice were perfused with 10 mM PBS (pH 7.4), and tumors were collected. Soft tissue and bonelike matrix compartments of the tumor were separated physically, weighed, and homogenized for phage recovery. The recovered phage pool was amplified and subjected to another round of selection. The two peptides, PKRGFQD and SNTRVAP, targeted the cell surface of androgen-independent prostate cancer cells in vitro, and homed to androgen receptor-null prostate cancer in vivo. Purification of tumor homogenates by affinity chromatography on these peptides and subsequent mass spectrometry revealed a receptor for the peptide PKRGFQD, α-2-macroglobulin, and for SNTRVAP, 78-kDa glucose-regulated protein (GRP78). 2816 SNTRVAP(11.7%) GAGPASV(2.4%) SNTFVAP(2.4%) RLGLAWG(2.1%) VTRGVGF(2.1%) 5 Phage display (in vivo) 3 PMID:25762070 Bonelike matrix tissues of prostate cancer xenograft Not determined. Not determined. Not determined. CX7C fUSE5 phage display library For round 1, tumor-bearing mice received 2e9 transducing units (TU) of phage peptide library i.v. After 24 h of circulation, the mice were perfused with 10 mM PBS (pH 7.4), and tumors were collected. Soft tissue and bonelike matrix compartments of the tumor were separated physically, weighed, and homogenized for phage recovery. The recovered phage pool was amplified and subjected to another round of selection. The two peptides, PKRGFQD and SNTRVAP, targeted the cell surface of androgen-independent prostate cancer cells in vitro, and homed to androgen receptor-null prostate cancer in vivo. Purification of tumor homogenates by affinity chromatography on these peptides and subsequent mass spectrometry revealed a receptor for the peptide PKRGFQD, α-2-macroglobulin, and for SNTRVAP, 78-kDa glucose-regulated protein (GRP78). 2817 GSPREYTSYMPH SLSSYNGSALAS 2 Phage display (in vivo) 3-4 PMID:25768046 The ligated left carotid artery in male C57/BL6J mice Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The ligated left carotid artery is the target for phage display, while the right artery serves as a normal flow control. Four days postligation, in vivo phage display was carried out. Phages were injected into the tail vein of the mice at a concentration of 2e11 plaque-forming units. After 2.5 h, mice were sacrificed by CO2 inhalation, and the carotid arteries were surgically isolated; phages were recovered from the vessels. Titration of phage normalized to the protein content of the corresponding LCA or RCA was used to determine the relative uptake between the target and control arteries. All four consensus clones showed a significantly increased binding to the targeted LCA. ACTPSFSKIC showed average enrichment of 7.3-fold; SLSSYNGSALAS showed enrichment of 4.9; ACNTGSPYEC showed enrichment of 6.1, and GSPREYTSYMPH showed enrichment of 4.4 in the target artery. 2818 CNTGSPYEC CTPSFSKIC 2 Phage display (in vivo) 3-4 PMID:25768046 The ligated left carotid artery in male C57/BL6J mice Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The ligated left carotid artery is the target for phage display, while the right artery serves as a normal flow control. Four days postligation, in vivo phage display was carried out. Phages were injected into the tail vein of the mice at a concentration of 2e11 plaque-forming units. After 2.5 h, mice were sacrificed by CO2 inhalation, and the carotid arteries were surgically isolated; phages were recovered from the vessels. Titration of phage normalized to the protein content of the corresponding LCA or RCA was used to determine the relative uptake between the target and control arteries. All four consensus clones showed a significantly increased binding to the targeted LCA. ACTPSFSKIC showed average enrichment of 7.3-fold; SLSSYNGSALAS showed enrichment of 4.9; ACNTGSPYEC showed enrichment of 6.1, and GSPREYTSYMPH showed enrichment of 4.4 in the target artery. 2819 WQRPSSW(2)[1.91, 1.80, 1.63] HLYWQRP(1)[1.28, 1.16, 0.98] 2 Phage display (competitive panning) 4 PMID:25771000 Human serum albumin Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Selected 7-mer peptides were tested for binding human serum albumin (HSA), rabbit serum albumin (BuSA) and mouse serum albumin (MSA) by ELISA. The OD405 values were determined. Data shown were reproduced from graph. After removing unbound phage by repeatedly washing the plate with TBST, bound phage were eluted with 1ml HSA solution (100 mg/ml). 2820 ICARQDPAGNCS[0.50] 1 Phage display (common panning) 3 PMID:25779283 Polyclonal anti-LiAg IgGs L. infantum protein antigen (LiAg) Not determined. Not determined. LX-8 phage display library (XCX8CX) ELISA Plate was sensitized with e10 phages/mL of individual clone and Wild type phage (negative control). Positive and negative dog serum were employed. Binding was detected using a peroxidase conjugated anti-dog IgG antibody. Absorbance values at 492 nm were measured. The A492 values of the phage peptide binding to positive and negative dog serum were 0.50 and 0.08, while for wild type phage the values were 0.10 and 0.06, respectively. Peptide ICARQDPAGNCS has a ability to promote a state of immunity against L. infantum infection in murine model after immunization using liposomes as vaccine carrier and is a promising vaccine candidate against CVL. 2821 ETTTEPNRTRASAPA(5) HYNTEPKARDLRTIP(1) AYYAEPKQKTYLRTP(1) HDYTEPRFTYQISNA(1) WTEPKGPNAPQDPWM(1) GELETEPRKNAGIMP(1) GPAPDGTEPRYYSRS(1) ERTRTEPKIKSIVSM(1) VSTEPKRPYAPTTGC(1) SGSTEPKRLNWDNVS(1) SEAEPHVHRLIYRPL(1) SPAEPRNERTAARMR(1) AAEPRRHLSNLPDTR(1) SVPQLEPKRPKHRIY(1) SELEPRRPRSPYPVT(1) DRRQTEPITMKYTEP(1) FTTEPPKAGRTYQVW(1) GQTEPPRQISFKVAP(1) ARPRIEPEKLRQLPG(1) RLATEPTHNGSHPAN(1) GAQLEPNKATTARLT(1) TMPSKHHFLDRAHPP(1) RTASVPSRDRTNYAP(1) Y-TEP, TEP 23 Phage display (common panning) 3 PMID:10712615 Anti-human cardiac troponin I mAb 11E12 Human cardiac troponin I (hcTnI) Not determined. Not determined. f88.4-based X15 phage display library 2822 LTWGEMHTWTVQ(20) VSWPELYKWTWS(16) TTFDILDYWTSN(4) ITTSEIYNWRDT(3) ITNAELTNWNNG(2) GQPWTTWLESNT(2) ITAPELYAWFGS(1) LTMEELTRWSVY(1) ITLPELHAWKEN(1) ITIQEITAWPES(1) LTNQELLTWTAY(1) MDLAELSNWPHA(1) LSIADLYRWNTS(1) WQIWEYWPMDHN(1) VTLGELVSWPAE(1) LTLEELLFWKSP(1) LTRLELLEWDSP(1) LSWEELLRWASP(1) LTHTELLHWNGM(1) ITQADVWAWDTS(1) 20 Phage display (common panning) 3 PMID:25785734 Anti-HIV-1 monoclonal antibody VRC01 Human immunodeficiency virus type 1, HIV-1 Not determined. Not determined. Ph.D.-12 phage display library (X12) 2823 CEWSLWSFC(6) CSWTLLGYC(5) CSWNLMGFC(5) CNWEFWKYC(5) CNWEFWKYC(5) CSWNLMGFC(5) CPWVLHGFC(4) CTWTLLSFC(4) CSWSLNGFC(3) CTFTYWGFC(1) CPWYLMGYC(1) CNWSLLSFC(1) CEWTWFGYC(1) CLWSLTGFC(1) CQWTYYNFC(1) CEWRYWEYC(1) CDWLLHGFC(1) CPWMLSGFC(1) CTWSLSGFC(1) CLWMLEKFC(1) CTHSRAGSC(1) CSWSLLDFC(1) CIWEFLGFC(1) 23 Phage display (common panning) 3 PMID:25785734 Anti-HIV-1 monoclonal antibody VRC01 Human immunodeficiency virus type 1, HIV-1 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2824 DCFVRCRVACA(2)[NT] SCFALCRVPCQ(2)[NT] KCFTACRVDCF(1)[NT] SCFNRCRVPCF(1)[NT] SCFDRCRVGGY(1)[NT] QCFELCRVVCL(1)[NT] RCFNLCRVGCL(1)[NT] VCFPLCRVPCI(1)[NT] ACFKACRVNCA(1)[7.5 ± 3.2] ACFKHCRVACA(1)[10.6 ± 0.5] DCFQGCRVFCS(1)[NT] MCFDSCRVNCT(1)[112.7 ± 22.1] TCFHRCRVTCI(1)[NT] KCFQACRASCY(1)[NT] RCFEKCRAFCY(1)[NT] VCFSLCRAGCL(1)[NT] VCFGVCRALCA(1)[NT] VCFQLCRAQCI(1)[450 ± 75.5] LCWTGCRVSCF(1)[NT] GCWAPCRVGCY(1)[NT] RCWTACRGLCF(1)[NT] RCWQTCRVSCV(1)[NT] QCWRSCRVNCL(1)[NT] GCYTRCRVDCF(1)[NT] VCYALCRVSCD(1)[NT] DCYQLCRVSCE(1)[NT] TCFRSCKVACY(1)[NT] QCFQACKTLCW(1)[NT] 28 Phage display (subtractive panning) 2 PMID:23100545 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX3CX3CX phage display library Inhibition assay The bicyclic peptides inhibited hPK with IC50s ranging from 7.5 to 450 nM. Bicyclic peptides were chemically synthesized and their inhibitory activity (IC50, nM) is indicated. NT denotes not tested. The library was joined and subjected to a first affinity selection with hPK. The second round of panning was performed using neutravidin-coated magnetic beads to prevent the enrichment of streptavidin-specific peptides. 2825 VCGTGWCQLARCI(1) 1 Phage display (subtractive panning) 2 PMID:23100545 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX4CX4CX phage display library The library was joined and subjected to a first affinity selection with hPK. The second round of panning was performed using neutravidin-coated magnetic beads to prevent the enrichment of streptavidin-specific peptides. 2826 WCFDVCRVGCL(2) VCFPLCRVPCI(2) GCYKPCRVNCP(2) DCFGRCRVRCL(2) NCFPLCRVGCE(1) GCFQECRVVCV(1) SCFQTCRVYCL(1) QCFERCRVNCD(1) DCFHQCRVGCD(1) KCWGLCRVGCL(1) VCWSPCRVACS(1) ACYEACRVNCV(1) SCYTGCRVYCI(1) TCDLVCQVRCR(1) RCPMSCNYRCV(1) KCGKKCDYRCL(1) TCEKRCCPRCS(1) 17 Phage display (subtractive panning) 2 PMID:23100545 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX3CX3CX phage display library The library was joined and subjected to a first affinity selection with hPK. Isolated phage were amplified and, prior to the second round of affinity selection, treated for 30 min with 5 mg/ml pancreatin. The second round of panning was performed using neutravidin-coated magnetic beads to prevent the enrichment of streptavidin-specific peptides. 2827 ECFQGFCWRDTCW(2) SCFSGFCWRQMCS(1) SCFQAACWRQLCG(1) RCERALCVWWWCD(1) SCGRDGCLWKSCR(1) 5 Phage display (subtractive panning) 2 PMID:23100545 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX4CX4CX phage display library The library was joined and subjected to a first affinity selection with hPK. Isolated phage were amplified and, prior to the second round of affinity selection, treated for 30 min with 5 mg/ml pancreatin. The second round of panning was performed using neutravidin-coated magnetic beads to prevent the enrichment of streptavidin-specific peptides. 2828 GCWPQCRVNCV(3)[115.7 ± 24] TCEKRCCPRCS(3)[NT] KCWQECRSLCY(2)[NT] GCYERCRVYCS(2)[NT] GCYKPCRVNCP(1)[158.7 ± 25.7] ECFTACRVACI(1)[NT] ECFDKCRVFCV(1)[NT] GCFTPCRTMCW(1)[NT] TCFHHCRVNCS(1)[NT] ACWKLCRVDCV(1)[NT] NCWARCRVMCE(1)[NT] FCFQNCKALCF(1)[NT] SCVRPCFYWCD(1)[NT] SCVDYCVGTCC(1)[NT] LCMDNCEQVCQ(1)[NT] KCGKKCDYRCL(1)[NT] 16 Phage display (subtractive panning) 2 PMID:23100545 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX3CX3CX phage display library Inhibition assay Bicyclic peptides were chemically synthesized and their inhibitory activity (IC50, nM) is indicated. NT denotes not tested. The library was joined and subjected to a first affinity selection with hPK. Isolated phage were amplified and, prior to the second round of affinity selection, treated for 30 min with 10 mg/ml pancreatin. The second round of panning was performed using neutravidin-coated magnetic beads to prevent the enrichment of streptavidin-specific peptides. 2829 SCFQAACWRQLCG(2) NCRAKGCWYGYCF(1) ACRSWSCTCGLCY(1) SCGGDQCALDRCV(1) TCSWGKCQVTDCG(1) 5 Phage display (subtractive panning) 2 PMID:23100545 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX4CX4CX phage display library The library was joined and subjected to a first affinity selection with hPK. Isolated phage were amplified and, prior to the second round of affinity selection, treated for 30 min with 10 mg/ml pancreatin. The second round of panning was performed using neutravidin-coated magnetic beads to prevent the enrichment of streptavidin-specific peptides. 2830 ACFDPCRVVCY(1)[18.2 ± 4.2] QCFQWCRTNCY(1)[178.3 ± 12.6] NCWNPCRVACL(1)[51.7 ± 10.4] QCENPCPTSCC(1)[NT] TCEKRCCPRCS(4)[NT] 5 Phage display (subtractive panning) 2 PMID:23100545 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX3CX3CX phage display library Inhibition assay Bicyclic peptides were chemically synthesized and their inhibitory activity (IC50, nM) is indicated. NT denotes not tested. The library was joined and subjected to a first affinity selection with hPK. Isolated phage were amplified and, prior to the second round of affinity selection, treated for 30 min with 20 mg/ml pancreatin. The second round of panning was performed using neutravidin-coated magnetic beads to prevent the enrichment of streptavidin-specific peptides. 2831 TCERRACVMRECA(1) GCVQGKCVLATCT(1) 2 Phage display (subtractive panning) 2 PMID:23100545 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX4CX4CX phage display library The library was joined and subjected to a first affinity selection with hPK. Isolated phage were amplified and, prior to the second round of affinity selection, treated for 30 min with 20 mg/ml pancreatin. The second round of panning was performed using neutravidin-coated magnetic beads to prevent the enrichment of streptavidin-specific peptides. 2832 CSTYNPYCQLKTSSC(7)[>75] CAQDEKSCWYTPLKC(4)[>75] CTAQNPYCALKQRSC(3)[NT] CTQGPGGCKDHLLLC(3)[NT] CPADQMLCKGLFCLC(3)[NT] CLSMERKCVKTQLCC(3)[NT] CSSLDASCWFSQTKC(2)[>75] CSRNGQDCFWSSIKC(2)[NT] CNRDTENCYWTQVKC(2)[NT] CLQNNYLCFPGTKPC(2)[NT] CAGANPYCLAKGQAC(2)[>75] CLGTPRKCTKTELCC(2)[NT] CTVKPSFCGNGGQAC(2)[NT] CSSFAQDCWFSYTKC(1)[NT] CTESPQTCWYTATKC(1)[NT] CFADTTECWFSITKC(1)[NT] CNSVNDMCWYSQTKC(1)[NT] CQGPSYDCFYSVSKC(1)[NT] CAQTDGGCYWSSVKC(1)[NT] CARDEQGCFYSVVKC(1)[>75] CQKKDEQCFWSAVKC(1)[>75] CSQNAPGCFYSKVKC(1)[NT] CVQGQADCFFSSAKC(1)[NT] CEDSWKTCWYSQVKC(1)[NT] CPGDVDSCFYSSVKC(1)[NT] CQPDEEGCWWNQVKC(1)[NT] CTIGHDECWYQTTKC(1)[NT] CGLATPDCWWNQTKC(1)[NT] CPTQGGECWQTQVKC(1)[NT] CYNAKETCWDSLTRC(1)[NT] CNTDQNLCYYSKSGC(1)[NT] CWWTSTKCRQEDNNC(1)[NT] CWQTVTKCGQSFVGC(1)[NT] CWVTQAKCPQASTEC(1)[NT] CWTTQVKCGGGAGDC(1)[NT] CDNINPYCVVKEQKC(1)[NT] CSEANPYCWLKPNQC(1)[NT] CNLIQPYCQLKDHMC(1)[NT] CPSLQPYCGVKWALC(1)[NT] CLLSSPYCAVKGQGC(1)[NT] CELRVPYCVVKNQRC(1)[NT] CIRLNPYCSQKFTMC(1)[NT] CSLNEKYCTQVKKPC(1)[NT] CEVPAVKCQFWGRGC(1)[NT] CQVPLAKCNWFLWPC(1)[>75] CQSWSRGCAKEILCC(1)[NT] CKIMNAYCAKTSLCC(1)[NT] CTNKQLFCPEKRQCC(1)[NT] CQDMNNICGSKAKFC(1)[NT] CQDWVGECFFTKIGC(1)[NT] CQAGNTACMFPAKHC(1)[NT] CQHAAEGCWHTIIKC(1)[NT] CQGKASFCEQLGHVC(1)[NT] CQGVMTLCFPQTKLC(1)[NT] CLQTADPCLKTPVKC(1)[NT] CLQNTEPCVYFATKC(1)[NT] CLQGVTVCGVKGTIC(1)[NT] CREQISKCFDGQEWC(1)[NT] CAEDLWGCIQLKSKC(1)[NT] CGQYTKFCFHNWNGC(1)[NT] CYKSLGTCQGRLSWC(1)[NT] [WF]-[FYW]-[ST]-x-[TV]-K 61 Phage display (subtractive panning) 3 PMID:23586812 Human alpha-FXIIa Not determined. Not determined. Not determined. CX6CX6C phage display library Inhibition assay Several bicyclic peptides presenting common sequence elements were chemically synthesized and their inhibitory activity (Ki, μM) is indicated. All indicated inhibitory constants Kis are averages of at least three measurements. NT denotes not tested. In the second round, neutravidin-coated magnetic beads were used to prevent the enrichment of streptavidin-specific peptides. 2833 GCVSDKCIIQYCN(5)[42 ± 4] ACVTGKCLLQLCP(3)[>75] ECVVDKCLVQFCP(3)[NT] QCVEDKCIQSLCA(3)[>75] NCVSQKCRWEYCP(3)[NT] DCVKPRCPQGGCR(3)[NT] LCKTQGCVSFSCE(3)[NT] YCVMDKCLQTVCT(2)[NT] HCFKAKCPIQGCW(2)[>75] TCVQGKCLITNCG(1)[NT] ECVQGKCLWSMCL(1)[49 ± 8] MCVQGKCLYLYCQ(1)[NT] GCVQGKCLQELCA(1)[NT] SCVQGKCLVSMCR(1)[NT] DCVVGKCLSQVCS(1)[NT] DCVVGKCLYDFCK(1)[NT] SCVVGKCLVQYCA(1)[NT] GCVIGKCLTYVCE(1)[NT] GCVSGKCLQIFCQ(1)[NT] DCVEGKCLQVNCL(1)[NT] SCVEGKCLAQFCS(1)[NT] GCVKGKCLQEFCP(1)[NT] LCVQDKCLPTRCP(1)[NT] FCVVEKCLAQQCQ(1)[NT] YCVREKCLPYNCY(1)[NT] SCVWDKCLQAICH(1)[NT] ACVAQKCLWDFCR(1)[NT] VCVMQKCLWNVCE(1)[NT] NCVQQKCLWSACL(1)[NT] ECVPVKCLWQTCA(1)[NT] QCVPSKCLYAMCN(1)[NT] ECVVGKCVWENCL(1)[NT] YCVSGKCRQEVCS(1)[NT] QCVFDKCIQNLCI(1)[NT] QCVFDKCIQNLCI(1)[NT] YCVLDKCQPLRCP(1)[NT] GCVPQHCGGPRCF(1)[NT] ACVQSLCNSVNCY(1)[NT] HCWLAKCPVQGCN(1)[NT] PCPIQLCGSSKCR(1)[NT] QCPLQLCGWVKCG(1)[NT] LCPDALCGSAKCM(1)[NT] LCPQEFCGSLKCG(1)[>75] DCQFSWCAVKRCP(1)[NT] GCQAFWCAVKQCY(1)[>75] GCQGSRCVKTLCC(1)[NT] WCQNGGCVKTLCC(1)[NT] GCQARACAKFLCC(1)[NT] GCGKKLCAKELCC(1)[NT] TCSEKRCSKLLCC(1)[NT] WCTHQRCVKARCC(1)[NT] RCKVAYCAKDLCC(1)[NT] RCGQGFCSYTKCS(1)[NT] SCSAGFCSPVKCP(1)[NT] GCWWGLCRYKECP(1)[NT] QCFSPYCMVKGCP(1)[NT] SCDYPWCQVKMCP(1)[NT] VCSPLKCGSAVCR(1)[NT] QCPKPRCQLVLCP(1)[NT] GCQNWACPVKPCH(1)[NT] K-C-[LI], AVK 60 Phage display (subtractive panning) 3 PMID:23586812 Human alpha-FXIIa Not determined. Not determined. Not determined. XCX4CX4CX phage display library Inhibition assay Several bicyclic peptides presenting common sequence elements were chemically synthesized and their inhibitory activity (Ki, μM) is indicated. All indicated inhibitory constants Kis are averages of at least three measurements. NT denotes not tested. In the second round, neutravidin-coated magnetic beads were used to prevent the enrichment of streptavidin-specific peptides. 2834 CSRTQDECWWTLTKC(42) CMQRTVACVKQTLCC(5) CTESPQTCWYTATKC(4) CSRNGQDCFWSSIKC(4) CSQGDLKCWWTLTKC(3) CDRVRPYCQVKGAAC(3) CFKGSSDCFYSQVKC(2) CNRDTENCYWTQVKC(2) CGMVNKRCVKELLCC(2) CIRYSFLCGAGGGEC(2) CMDEVWMCFGITGLC(2) CGISSRSCWYTITKC(1) CEGTDSQCWYSIVKC(1) CQGMVDTCFWSALKC(1) CDQSMQACFWSSIKC(1) CQGPSYDCFYSVSKC(1) CYKGPVSCFWNPTKC(1) CQKGPVSCFWNPTKC(1) CQGGQENCWLNPTKC(1) CQDGGYSCFFTKTGC(1) CQLVYPYCRVKVENC(1) CVYKMPYCYVKQLNC(1) CSTYNPYCQLKTSSC(1) CMYRAPYCQIKQTSC(1) CEAVNPYCQRKGRSC(1) CPADQMLCKGLFCLC(1) CEWHQKICKGTFCLC(1) CQEMKWWCPSVKYHC(1) CSQVHGWCPSAKYGC(1) CTKGQRSCAKLSLCC(1) CDAFSRGCVKQHMCC(1) CMSRMATCVKASRCC(1) CGFMRHSCSKTTLCC(1) CGGFRWCCDISKIRC(1) CSGQRWCCDVRSARC(1) CSSSRWCCSYQSVLC(1) CLRTLGSCGSTMLAC(1) CYHENAYCKRKWGDC(1) CSGKWLYCGQSAKLC(1) CLKCAGACPFKGQAC(1) CRQDWWVCEWSWSTC(1) CVSPRQMCTWPKSNC(1) CDKGTRKCGYLRFEC(1) [WF]-[WY]-[TS]-x-[TVL]-K, PYCQ 43 Phage display (subtractive panning) 3 PMID:23586812 Human beta-FXIIa Not determined. Not determined. Not determined. CX6CX6C phage display library In the second round, neutravidin-coated magnetic beads were used to prevent the enrichment of streptavidin-specific peptides. 2835 ACDARPCPQTYCL(28)[20.5 ± 5.2] SCGGRPCPPAYCK(22)[3.1 ± 0.5] GCLGRPCPMAYCS(13)[5.0 ± 1.5] DCPVVWCLQKACR(7)[46.9 ± 3.2] VCVQKFCWRGWCP(4)[>75] QCNARPCPSSYCR(2)[4.7 ± 1.5] GCMGRPCPVSYCF(2)[5.0 ± 1.3] ACQQQFCWRGWCP(2)[NT] LCEYTLCWRGWCP(2)[NT] HCRYVFCWRGWCP(2)[NT] WCVMQKCLMQYCE(2)[NT] YCVKDKCLQAMCS(2)[NT] SCGGPRCWPSTCR(1)[NT] RCGQGFCSPVKCR(1)[NT] ICKSQVCVSFSCG(1)[NT] GCWARPCPLALCQ(1)[10.2 ± 4.6] GCAARPCPLTACW(1)[33.5 ± 5.9] GCHGRPCPLQYCK(1)[11.2 ± 4.4] RCYANPCPISYCR(1)[NT] SCSGRRCPPSYCK(1)[7.8 ± 3.2] NCVNRYCWRGWCS(1)[NT] SCVVGKCLVQYCA(1)[NT] HCVQGKCLVYMCG(1)[NT] QCVPFKCLLHLCA(1)[NT] GCVLGKCLQDFCQ(1)[NT] YCVPLKCLWDRCE(1)[NT] RCSAGGCCERVCE(1)[NT] TCTTQGCAFRLCS(1)[>75] WCTHQRCVKARCC(1)[NT] YCQGTKCVKERCC(1)[NT] LCQPRKCVKALCC(1)[NT] SCANQRCVKSLCC(1)[NT] TCLCKRCIKELCC(1)[NT] VCTYRKCVKELCC(1)[NT] GCQARACAKFLCC(1)[NT] GCGRNICVKNLCC(1)[NT] [AG]-R-P-C-P, WRGW, V-K-x-[RL] 36 Phage display (subtractive panning) 3 PMID:23586812 Human beta-FXIIa Not determined. Not determined. Not determined. XCX4CX4CX phage display library Inhibition assay Several bicyclic peptides presenting common sequence elements were chemically synthesized and their inhibitory activity (Ki, μM) is indicated. All indicated inhibitory constants Kis are averages of at least three measurements. NT denotes not tested. In the second round, neutravidin-coated magnetic beads were used to prevent the enrichment of streptavidin-specific peptides. 2836 CDSRFRNCPWSMSGC[1500] CSDRFRNCPLWSGTC[400] CRDIRFRCNYDVAVC[NT] CSTERRYCPIEIFPC[400] CAPWRTMCLNIDGPC[NT] CAPWRTACYEDLMWC[500] CPVWRTLCMESEGVC[NT] CFPLWRTCVHEPTMC[NT] CWQVQVNCRVNFGKC[800] CGGNSDRCRVNNISC[2000] CGRGDQTCRVNWHPC[NT] CGTGEGRCRVNWTPC[500] APWRT, RFRN, RVN 12 Phage display (subtractive panning) 3 PMID:19483697 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. CX6CX6C phage display library Inhibition assay The IC50 (nM) was measured by incubating various concentrations of the modified peptide fusion proteins with human plasma kallikrein (0.1 nM) or cathepsin G (20 nM). In the second round of selection, neutravidin-coated magnetic beads were used to prevent the enrichment of streptavidin-specific peptides. 2837 CEYGDLWCGWDPPVC[NT] CEYDVGFCWDGFGDC[100] CLFDAGFCQQHSTEC[NT] CIFDLGFCHNDWWNC[100] CLFDLGFCGGGEGPC[150] CPRIEGFCLPIFSDC[1000] CLRAQEDCVYDRGFC[200] CTRGSGDCTYDFGFC[200] LFD 8 Phage display (subtractive panning) 3 PMID:19483697 Cathepsin G (EC:3.4.21.20) Not determined. Not determined. Not determined. CX6CX6C phage display library Inhibition assay The IC50 (nM) was measured by incubating various concentrations of the modified peptide fusion proteins with human plasma kallikrein (0.1 nM) or cathepsin G (20 nM). In the second round of selection, neutravidin-coated magnetic beads were used to prevent the enrichment of streptavidin-specific peptides. 2838 CNERFSMCGQMGLRC(5)[NT] CNAKFSGCRYSALVC(2)[3.66 ± 0.09] CNSRFSGCQIDLLMC(2)[NT] CNAKFSGCVGRGGHC(1)[NT] CNAKFSLCGNSVFGC(1)[NT] CNAYFSGCNSLTGGC(1)[NT] CNAYFSGCDWAVQHC(1)[2.638 ± 0.081] CNARFSGCQPSPAAC(1)[NT] CNNKFTLCGSRQIIC(1)[NT] CNWKFSLCETQRNQC(1)[NT] CNSKFTGCRAGTGLC(1)[11.136 ± 0.294] CNSKFTQCGAPRGTC(1)[NT] CNSRFALCSPSSQMC(1)[NT] CNSKYSGCQRSPASC(1)[NT] CNSKYSGCGGQRYDC(1)[NT] CNSRYSLCTGAILTC(1)[12.341 ± 0.331] CNERYALCGILGFQC(1)[NT] CSRYEVDCRGRGSAC(1)[0.071 ± 0.004] CAEQVIDCRGRGGLC(1)[NT] N-[AS]-[KRY]-[FY]-[ST], DCRGRG 19 Phage display (subtractive panning) 2 PMID:22304751 Urokinase-type plasminogen activator (EC:3.4.21.73) Not determined. Not determined. Not determined. CX6CX6C phage display library Competition experiment The inhibitory activity of bicyclic peptides was determined by incubating human uPA with different concentrations of inhibitor and quantification of the residual activity with a fluorogenic substrate. The inhibitory activities (Ki, μM) of five TBMB-cyclized peptides (as D1-D2 fusion peptides) are indicated. A second round of panning was performed following the same procedure but using neutravidin-coated magnetic beads in order to prevent the enrichment of streptavidin-specific peptides. 2839 AAKTPTE(8) ATTVPAS(4) MSLQQEH(4) SAPSSKN(2) LPRTPTD(2) MHAPPFY(1) DEQPYDL(1) TSLSMPS(1) 8 Phage display (common panning) 3 PMID:25663979 Visfatin Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2840 ICARQDPAGNCS[1.54] KCPSIPGAVLCV[1.57] 2 Phage display (common panning) 3 PMID:25710003 Polyclonal anti-LiPA IgG L. infantum chagasi Protein Antigen (LiPA) Not determined. Not determined. LX-8 phage display library (XCX8CX) ELISA Individual clones isolated was added to each well of microtiter plates coated with 1 μg/well of anti-LiPA IgG and anti- T. cruzi IgGs. Phages were detected using a peroxidase conjugated anti-M13 antibody (1 : 3000). Values of absorbance at 492 nm are measured. The A492 values of phages with ICARQDPAGNCS and KCPSIPGAVLCV insert for anti- T. cruzi IgGs were 0.19 and 0.19, respectively. Data shown were reproduced from graph. Anti-LiPA IgGs were coated onto a polystyrene Petri dish overnight at a concentration of 5 μg/mL for the first two rounds of panning and a concentration of 0.5 μg/mL for the last panning. 2841 TTDDDKLKKTLTYRS[1.51] 1 Phage display (common panning) 3 PMID:25710003 Polyclonal anti-LiPA IgG L. infantum chagasi Protein Antigen (LiPA) Not determined. Not determined. X15 phage display library ELISA Individual clones isolated was added to each well of microtiter plates coated with 1 μg/well of anti-LiPA IgG and anti- T. cruzi IgGs. Phages were detected using a peroxidase conjugated anti-M13 antibody (1 : 3000). Values of absorbance at 492 nm are measured. The A492 value of phages with TTDDDKLKKTLTYRS insert for anti- T. cruzi IgGs was 0.18. Data shown were reproduced from graph. Anti-LiPA IgGs were coated onto a polystyrene Petri dish overnight at a concentration of 5 μg/mL for the first two rounds of panning and a concentration of 0.5 μg/mL for the last panning. 2842 CFKACRVNC[5.2 ± 1.8] CFKQCRVNC[NT] CFKNCRVNC[NT] CFYKCRVNC[NT] CFNKCRVNC[NT] CFPKCRVAC[NT] CFKHCRVAC[5.2 ± 1.9] CFDPCRVIC[NT] RVN 8 Phage display (subtractive panning) 3 PMID:22492508 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. CX3CX3C phage display library Competition experiment Target specificity of bicyclic peptides was indicated by the Ki values (nM) of these peptides against hPK, monkeyPK and rPK. The Ki values of one peptide CFKACRVNC against monkeyPK and rPK were 2.8 ± 1.7 and 22.7 ± 14.0, and for the other peptide CFKHCRVAC, the corresponding values were 4.4 ± 1.1 and 14.0 ± 8.2. In the second round of selection, neutravidin-coated magnetic beads were used to prevent the enrichment of streptavidin-specific peptides. 2843 SCFNRCRVPCF[NT] KCFTACRVDCF[NT] RCFNLCRVGCL[NT] DCFQGCRVFCS[NT] QCFELCRVVCL[NT] MCFDSCRVNCT[NT] SCFALCRVPCQ[NT] VCFPLCRVPCI[37.1 ± 8.1] VCFGVCRVLCA[NT] KCFQACRVSCY[NT] VCFSLCRAGCL[NT] RCFEKCRAFCY[NT] RCWQTCRVSCV[NT] LCWTGCRVSCF[NT] GCWAPCRVGCY[NT] RCWTACRGLCF[NT] DCYQLCRVSCE[NT] TCFRSCKVACY[NT] QCFQACKTLCW[NT] 19 Phage display (subtractive panning) 3 PMID:22492508 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX3CX3CX phage display library Competition experiment Target specificity of bicyclic peptides was indicated by the Ki values (nM) of these peptides against hPK, monkeyPK and rPK. The Ki values of the peptide VCFPLCRVPCI against monkeyPK and rPK were 12.2 ± 7.4 and 59.7 ± 22.4 respectively. In the second round of selection, neutravidin-coated magnetic beads were used to prevent the enrichment of streptavidin-specific peptides. 2844 CAWPARCLTVDLC[0.4 ± 0.1] CRWPARCVHQDLC[0.3] CRWPARCLTTSLC[NT] CAWPAKCLTRELC[NT] CAWPARCLTTSLC[0.9 ± 0.1] CSWPARCNHQDLC[NT] CRWPARCTHQNYC[2.9 ± 1.2] CTWPARCTHQNWC[NT] CTWPARCTMQNYC[NT] CSWPYRCLHQDYC[NT] CNWPYRCLHTDLC[NT] CGVPYRCTHQEMC[NT] CTYPYKCLHQNLC[12.2 ± 4.8] CADPWACLFRRPC[NT] CGGPQNCRTWTTC[4.3 ± 0.2] CGGYNNCRAFSYC[NT] CFPSHDCDGRRMC[5.3 ± 2.8] PAR 17 Phage display (subtractive panning) 3 PMID:22492508 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. CX5CX5C phage display library Inhibition assay Target specificity of bicyclic peptides was indicated by the Ki values (nM) of these peptides against hPK. In the second round of selection, neutravidin-coated magnetic beads were used to prevent the enrichment of streptavidin-specific peptides. 2845 IGLTRY[0.36] DGSSKL[0.32] TLVSSL[0.30] LPAKSP[0.27] ALGRFQ[0.26] SIFPPK[0.26] LASNRQ[0.21] FRLIRS[0.20] LDAMVS[0.19] PWAWTS[0.19] WYTAPT[0.17] MVKWGT[0.15] SLWRLS[0.15] PLGVMR[0.14] HGSTEV[0.13] KFAGVN[0.13] HPTQST[0.12] AEKVLH[0.11] NNGGLT[0.09] SSRIGF[0.08] AWALRF[0.07] HPTGSM[0.06] 22 Phage display (subtractive panning) 7 PMID:16760366 Phosphatidylserine, PS Not determined. Not determined. Not determined. X6 fUSE5 phage dislpay library ELISA Phages bound to PS-coated plates were detected by incubation with horseradish peroxidase (HRP)–conjugated anti-M13 phage antibody followed by addition of the o-phenylenediamine dihydrochloride (OPD) substrate. Accumulation of the reaction product was detected by absorbance measured at 450 nm (A450 nm). Data shown were reproduced from graph. Four and three consecutive panning rounds of the phage library were performed, respectively, on control livers and on apoptotic livers. Four selected phages (IGLTRY, DGSSKL, TLVSSL, LPAKSP) were further evaluated for binding to PS-coated enzyme-linked immunosorbent assay (ELISA) plates. They presented an apparent affinity constant (Ka) for PS ranging from 6.08e10 M to 1.62e11 M. These phages did not bind to phosphatidylcholine, and competition with annexin V confirmed their specific interaction with PS. 2846 DAHSFS(5) DAHSLS(1) PGDLST(7) PGDLSR(3) PGDLVR(1) HGDLST(1) HGHLSI(1) VLGERG(1) LIKKPF(1) 9 Phage display (subtractive panning) 4 PMID:19743879 Phosphatidylserine, PS Not determined. Not determined. Not determined. X6 fUSE5 phage dislpay library The libary was incubated (2 h, room temperature) with a preblocked well (in the absence of PS) to remove phages that interact with plastic surface. And then, libary was transferred to the PS coated well and incubated overnight at room temperature under mechanical agitation. 2847 ARLHTSS(7) SMTKIMC(5) NGPFGKC(1) LDYKHYC(1) DYHLKPC(1) AACVQTC(1) KIPRVFC(1) MSVSKPC(1) AQHRTHL(1) 9 Phage display (common panning) 3 PMID:25469678 Anti-buffalo β-lactoglobulin antibody from rabbit 1 Beta-lactoglobulin, Beta-LG Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2848 SMTKIMC(3) IPLVVPF(2) AACVQTC(2) KIPAVFC(2) SEHGRNC(1) WMEQNGC(1) SVKPVRC(1) GIDWLYC(1) GIDWLYC(1) TQRGPAC(1) MTSVMMC(1) DYHLKPC(1) 12 Phage display (common panning) 3 PMID:25469678 Anti-buffalo β-lactoglobulin antibody from rabbit 2 Beta-lactoglobulin, Beta-LG Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2849 WHTGTPH(11) LHKHPTP(3) VHPHWRH(1) WHSGTPH(1) 4 Phage display (subtractive panning, competitive panning) 4 PMID:25565805 Mycobacterium tuberculosis (H37Rv) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the first round of biopanning, inactive MTB reference strain H37Rv was used as the target. For three further rounds of biopanning, Mycobacterium smegmatis was used as the reverse screening molecule with increased stringency of the washing steps. After four rounds of screening, Clones were eluted by H37Rv. 2850 WHTGTPH(4) VHPHWRH(4) LHKHPTP(3) CHPHTPH(1) 4 Phage display (subtractive panning, competitive panning) 4 PMID:25565805 Mycobacterium tuberculosis (H37Rv) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) In the first round of biopanning, inactive MTB reference strain H37Rv was used as the target. For three further rounds of biopanning, Mycobacterium smegmatis was used as the reverse screening molecule with increased stringency of the washing steps. After four rounds of screening, Clones were eluted by Mycobacterium smegmatis. 2851 CSAFLKE(1) VSACEKF(1) 2 Phage display (common panning) 4 PMID:24341921 B3β2γ Not determined. Not determined. Not determined. Acid-pp M13 phage display library (XSAX2KX) 2852 ISACEKF(2) CSAFLKD(1) 2 Phage display (common panning) 5 PMID:24341921 B3β2γ Not determined. Not determined. Not determined. Acid-pp M13 phage display library (XSAX2KX) 2853 CSAFLKE(3) CSAFLKD(2) VSALLKD(1) 3 Phage display (common panning) 6 PMID:24341921 B3β2γ Not determined. Not determined. Not determined. Acid-pp M13 phage display library (XSAX2KX) 2854 ASASQKS(1) QSAVYKA(1) LSACNKV(1) TSASLKV(1) GSAPPKS(1) 5 Phage display (common panning) 4 PMID:24341921 B3β2γ-variant1 Not determined. Not determined. Not determined. Acid-pp M13 phage display library (XSAX2KX) 2855 RSAVHKQ(1) 1 Phage display (common panning) 5 PMID:24341921 B3β2γ-variant1 Not determined. Not determined. Not determined. Acid-pp M13 phage display library (XSAX2KX) 2856 TSARLKL(1) TSARAKQ(1) 2 Phage display (common panning) 6 PMID:24341921 B3β2γ-variant1 Not determined. Not determined. Not determined. Acid-pp M13 phage display library (XSAX2KX) 2857 DWERPTPYELSI(7)[+++] TPRPTQYWLHL(1)[++++] SGLLPTPQTRPV(1)[+++] WPTSWEVTASFN(1)[++] DHARPTSVWLSR(1)[+] HGISEMWYLTPK(1)[+] 6 Phage display (common panning) 3 PMID:11750040 Anti-NTX monoclonal antibody BNTX21 BAG family molecular chaperone regulator 1 Not determined. Not determined. Ph.D.-12 phage display library (X12) Western blot +, relative reactivity with mAb BNTX21 obtained in immunoblot assays. 2858 NALWMPT(1)[+++] SANMPTN(1)[+++] YLGLTPM(1)[++] NRLWMPS(1)[++] NKVWPPS(1)[++] TSYTGPA(1)[++] NKAWFPN(1)[+] 7 Phage display (common panning) 3 PMID:11750040 Anti-NTX monoclonal antibody BNTX21 BAG family molecular chaperone regulator 1 Not determined. Not determined. Ph.D.-7 phage display library (X7) Western blot +, relative reactivity with mAb BNTX21 obtained in immunoblot assays. 2859 ACVWSWGWCQPTCGSCG(2) ACEPKDFSCVQDDHPCG(2) ACCWSEQVCHPQGDWCG(1) ACGGWLGCCHPQDPSCG(1) ACLWDAMCCFQDDHPCG(1) ACWQASQCCYFPDDWCG(1) ACHPQQGNCCYQTLPCG(1) ACHPQGDHCCENRESCG(1) ACNLWDPLCCHPQNTCG(1) ACESQFEWCFCNLQPCG(1) ACDNRWQTCSWWCWDCG(1) ACQVGYTNCRTPYCGCG(1) ACHPQDGRCQSLFECCG(1) ACVHGLWPCHPQGDQCG(1) ACQDWAPYCELLNHPCG(1) ACLVSDMRCPHPQAPRG(1) AVGQDLSCFSNTHPCGG(1) HPQ 17 Phage display (common panning) 2 PMID:23560397 Streptavidin Biotin Not determined. Not determined. CX6CX6C phage display library Phage of the library were cyclized by oxidation and then subjected to affinity selections with the model target streptavidin. For the selection of binders to streptavidin, the streptavidin beads (50 μL) were directly blocked. The HPQ motif was present in different positions of the peptides, preferentially following one of the cysteine residues. 2860 ACGLQPMACHPQNDNCG(1) ACVSWHGGCHPQGDICG(1) ACVQNLFTCHPQNAYCG(1) ACQPIQQDCQWHPQGCG(1) ACQQHPQNCTRTEGNCG(1) ACYWGNDLCHPQNTGWG(1) HPQ 6 Phage display (common panning) 2 PMID:23560397 Streptavidin Biotin Not determined. Not determined. CX6CX6C phage display library Phage of the library were cyclized by reaction with TBMB and then subjected to affinity selections with the model target streptavidin. For the selection of binders to streptavidin, the streptavidin beads (50 μL) were directly blocked. The HPQ motif was present in different positions of the peptides, preferentially following one of the cysteine residues. 2861 CCRERGCENMVCP(8)[4.3] CCLGRGCENHRCL(4)[7.7] MCNAYFAGDLC(2)[NT] LFQGACDTGRCA(2)[NT] CCREPPCYNPLCI(1)[73.8] CCGQAPCYLPACG(1)[299.7] WCSCTGCENGGCR(1)[82.7] SCGCRGCENRFCA(1)[118.8] QCPMDCCSRGLCW(1)[67.1] GCPDDCCSRGLCL(1)[50.3] YCSMDPCGTGRCR(1)[444.7] NCKYTLCSGVLW(1)[>1000] NCLYSGCLQVGCC(1)[>1000] YCNTARCGQAYCL(1)[>1000] CCRERGCEGQVCP(1)[NT] CCSERGCENVFCG(1)[NT] CCQGRGCENTWCV(1)[NT] CCLDRSCDGLMCI(1)[NT] GCCVSGCGYMSCG(1)[NT] YCPQDCCSRHLCM(1)[NT] SCVRGGCCGSACG(1)[NT] VCPQDGCAVCPCR(1)[NT] NCLFAGCLGSMCC(1)[NT] NCLFSACQGLTCC(1)[NT] NCTARSCASPDCC(1)[NT] GCGTGRCGQVWCT(1)[NT] VCVTARCQLQWCL(1)[NT] VCRLSYCSARTCP(1)[NT] DCLVTYCPQVRCQ(1)[NT] LCALRGCENRSCS(1)[NT] NCKYSLSASSDCQ(1)[NT] VCSIYFALGC(1)[NT] RCISSARMGTLCS(1)[NT] VCAVGGRLQENCL(1)[NT] RCDRCR(1)[NT] GCSVYFTCQ(1)[NT] R-[ED]-R-G-C-[ED]-N-x-V, P-C-Y-x-P, RGCEN, P-x-D-C-C-S-R-x-[LV], L-[YF]-S-G-C-L, TARCGQ 36 Phage display (common panning) 2 PMID:23560397 Urokinase-type plasminogen activator Not determined. Not determined. Not determined. XCX4CX4CX phage display library Inhibition assay The inhibition constants (Ki, μM) were determined by incubating different concentrations of bicyclic peptides with fixed concentration of human uPA (1.5 nM). NT denotes not tested. For the uPA selections, neutravidin-coated magnetic beads were used to immobilize the biotinylated human uPA. 2862 TCGVQACLSARCY(2)[12.9] NCRFSGCLQTMCV(2)[8.8] NCRFSGCGTVACV(1)[10.9] GCSDQACWSARCV(1)[10.5] NCKFSGCSVSVCH(1)[5.1] NCKFSGCPWELCI(1)[0.8] DCRWSSCTARTCA(1)[8.6] NCRFSGCVWAKCS(1)[NT] NCKFSGCQFLGCE(1)[NT] SCKFSGCHRKPCT(1)[NT] NCKFSACYLTTCY(1)[NT] NCTFTLCGLAPCG(1)[NT] SCLFTFCYFVPCN(1)[NT] NCRYSGCFDTPCI(1)[NT] GCQTSQCTARTCL(1)[NT] GCTESSCSARTCP(1)[NT] VCRQSDCSRRTCI(1)[NT] ECTASDCSARVCW(1)[NT] RCLPSWCSARTCF(1)[NT] RCLLSACTARACN(1)[NT] GCAQSVCTARYCE(1)[NT] ICSVLGCLSARCG(1)[NT] GCSVQACFSGRCT(1)[NT] PCGEQACYTARCR(1)[NT] LCLSQACWSARCA(1)[NT] VCVERGCSTARCR(1)[NT] LCADVSCATARCR(1)[NT] ECQDRSCPFTMCS(1)[NT] NCYFSNCGAHSAE(1)[NT] N-C-[RK]-F-S-G, R-x-S-x-C-[TS]-A-R-T, S-V-Q-A-C-x-[ST]-A-R 29 Phage display (common panning) 2 PMID:23560397 Urokinase-type plasminogen activator Not determined. Not determined. Not determined. XCX4CX4CX phage display library Inhibition assay The inhibition constants (Ki, μM) were determined by incubating different concentrations of bicyclic peptides with fixed concentration of human uPA (1.5 nM). NT denotes not tested. For the uPA selections, neutravidin-coated magnetic beads were used to immobilize the biotinylated human uPA. 2863 LPLGPHT[0.066 ± 0.002] TDHSMPP[0.155 ± 0.002] SSPPKSP[0.131 ± 0.002] 3 Phage display (common panning) 4 PMID:25515813 Fibroblast growth factor 23(180-205) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Affinities of the positive phage clones to FGF23 were measured by phage ELISA. The OD values were measured at 450 nm with a reference wavelength of 655 nm and data shown were reproduced from Fig. 1 in the reference. The affinity value of control phage vcsM13 to FGF-23 is 0.002. The affinity value was presented as mean ± standard deviation. After four rounds of panning, three positive clones with higher affinity and specificity for FGF23 were selected from 20 clones detected and further subjected to sequencing. 2864 AAAPLAQPHMWA(4)[11.50 ± 0.56, 13.7] SSTTTSDKYLSA(2)[9.31 ± 1.46, 10.8] SNLHDNNTEKNV(1)[8.06, 9.3] 3 Phage display (common panning) 4 PMID:25826583 Tumor necrosis factor ligand superfamily member 13 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Phage ELISA was used for identification of specific sAPRIL-binding phage. Clone 21 (sequence not shown) was used as a positive control. The fold change of the optical density was normalized to the positive control. Clones with an A450 that was at least 6 fold higher than the positive control were considered positive for sAPRIL binding. Besides, three binding peptides were synthesized and their binding affinity with sAPRIL was determined and compared with an unrelated polypeptide (HWDPFSLSAYFP) as a negative control using peptide ELISA. Data in the first column of affinity values were results for phage ELISA and reproduced from Fig. 1A in the reference. Data in the second column of affinity values were results for peptide ELISA and reproduced from Fig. 1B in the reference. The sAPRIL-BP1 (AAAPLAQPHMWA) had the highest binding affinity and was subsequently used in vitro to assess whether it was able to inhibit sAPRIL binding in the fixed human colorectal cancer cell line LOVO cells. sAPRIL-BP1 showed a dose-dependent inhibitory effect on sAPRIL binding to the LOVO cells. In vivo in a mouse colorectal challenge model, the sAPRIL-BP1 reduced the growth of tumor xenografts in nude mice by inhibiting proliferation and inducing apoptosis intratumorally. Moreover, in an in vivo metastasis model, sAPRIL-BP1 reduced liver metastasis of colorectal cancer cells. It might be a candidate for treating colorectal cancers that express high levels of APRIL. 2865 CTVRTSADC(5/29)[11-384] 1 Phage display (common panning) 4 PMID:25848940 Extradomain-B fibronectin (EDB-FN) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Surface plasmon resonance (SPR) SPR was performed characterize the interaction between CTVRTSADC and EDB using Biacore T100 (GE healthcare, Waukesha, WI). The binding data was analyzed using the Scatchard plot. Affinity (Kd, μM) was calculated by fitting data points into a linear trend line using Microsoft Office Excel software. Two binding sites were identified by fitting data from 1.9 μM to 7.8 μM or data from 15.6 μM to 250 μM. A tight binding site had an affinity of 11 μM, and a weak site was measured to have an affinity of 384 μM. A cyclic nonapeptide, CTVRTSADC (ZD2), was identified. Then a ZD2-Cy5 conjugate was synthesized to accomplish molecular imaging of prostate cancer in vitro and in vivo. ZD2-Cy5 demonstrated effective binding to up-regulated EDB-FN secreted by TGF-β-induced PC3 cancer cells following EMT. Following intravenous injections, the targeted fluorescent probe specifically bound to and delineated PC3-GFP prostate tumors in nude mice bearing the tumor xenografts. ZD2-Cy5 also showed stronger binding to human prostate tumor specimens with a higher Gleason score (GS9) compared to those with a lower score (GS 7), with no binding in benign prostatic hyperplasia (BPH). Thus, the ZD2 peptide is a promising strategy for molecular imaging and targeted therapy of prostate cancer. 2866 LPADEDHHFYWT(8)[2.85 ± 0.33][12.82 ± 1.73] LLQLTTTKRPIT(7)[5.80 ± 0.52][22.39 ± 2.30] QTHHTHLPIQRA(5)[3.54 ± 0.25][11.46 ± 0.42] TINSNTLNSTPP(5)[5.36 ± 0.99][10.10 ± 0.31] QKESSTHFMAIH(3)[3.61 ± 0.86][NT] SFPTMTENFYPR(3)[3.18 ± 0.63][NT] MHDASPQLYRGR(2)[4.28 ± 0.85][NT] MEDNDTHWARMA(2)[3.07 ± 0.13][NT] TMYEPTTTRSPA(1)[2.72 ± 0.28][NT] SVTAGMPNRSNR(1)[3.85 ± 0.76][NT] HLSVAKKPLHRP(1)[2.20 ± 0.24][NT] WPGLLGTTSPNI(1)[2.32 ± 2.46][NT] VHWPASQPVQIP(1)[2.24 ± 0.26][NT] HDHNTNDMTLNA(1)[2.70 ± 0.08][NT] 14 Phage display (subtractive panning) 4 PMID:25886968 Esophageal squamous carcinoma cell Eca-109 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA,Fluorescence Evaluation of the binding selectivity of 60 phage clones were performed by cellular ELISA. Eca-109 and HEK293 cells were used in ELISA. The 96-well plates were measured at 450 nm using an ELISA reader (Bio-Tek ELX800, USA). Irrelevant phage clone (IRP, an amplified phage randomly selected from the original phage peptide library) and PBS were used as control groups. Binding was showed as a ratio of cancer/normal cell binding. Data in the first column of affinity values were reproduced from Fig. 1 in the reference and expressed as mean ± the standard deviation of three replicates. The values for IRP and PBS are 1.25 ± 0.10 and 1.05 ± 0.30, respectively. In addition, cell immunofluorescence assay was performed. Fluorescence images were observed using laser scanning confocal microscope (LSCM, Leica, Germany). Fluorescence signal intensity (IOD/Area) in the second column of affinity values was reproduced from Fig. 2B in the reference. Fluorescence signal intensity for IRP and PBS controls is 3.66155 ± 0.21256 and 2.04334, respectively. NT denotes not tested. Phages were firstly incubated with HEK293 cells. After incubation, the supernatant containing unbound phages was incubated with the blocked Eca-109 cells. The 60 randomly selected phage clones were tested using cellular enzyme-linked immunosorbent assay (ELISA), and 41 phage clones were identified as positive clones with the over 2.10 ratio of absorbance higher than other clones, IRP and PBS controls. From the sequencing results of the positive clones, 14 peptide sequences were obtained and LLQLTTTKRPIT (ESCP9) consensus sequence was identified as the peptide with best affinity to ESCC cells via competitive inhibition, fluorescence microscopy, and flow cytometry. The results indicate that the peptide ESCP9 can bind to ESCC cells specifically and sensitively, and it is a potential candidate to be developed as an useful molecule to the imaging detection and targeting therapy for ESCC. 2867 QTTHRNWA (8) HTSHRNWD (4) FPHPWNPP (3) YPHPWNPT (3) PRYLPWFP (2) 5 Phage display (subtractive panning) 3 PMID:23394544 Anti-aflatoxin monoclonal antibody 1C11 Aflatoxin B1 Not determined. Not determined. CX8C M13 phage display library To eliminate nonspecific binding of phage with BSA, phages were firstly incubated with plate coated with BSA-PBS. After incubation, the supernatant containing unbound phages was incubated with the the plate containing 1C11 MAb. To elute the bound phage, 100 μL AFB1 (100 ng/mL in 10% methanol, competitive elution) was added to each well with shaking for 30 min to compete the binding phage from the coating antibody. 2868 LHSKHTYE(12)[13.9, 0.174] PTKAGIQS(5)[28.7, 0.087] PNPKSKTI(3)[44.3, 0.055] 3 Phage display (subtractive panning) 3 PMID:25908560 Anti-thiacloprid monoclonal antibody 3A5 Thiacloprid Not determined. Not determined. CX8C M13 phage display library Competition experiment The evaluations of the phage-displayed peptides were based on the IC50 (ug/L) and Amax/IC50 (the ratio of Amax to IC50), the combination of low IC50 and high Amax/IC50 was the most desirable. To eliminate nonspecific binding of phage to BSA, phages were firstly incubated with plate coated with BSA-PBS. After incubation, the supernatant containing unbound phages was incubated with the the plate containing 3A5 mAb. To elute the bound phage using competitive elution, 100 μL of thiacloprid (10 mg/L in PBS) was added to each well with shaking for 1 h to compete the binding phage from the coating antibody. 2869 LEIRSTVV(8)[55.7, 0.044] EYQYQTGL(7)[77.9, 0.031] AKSLEIDQ(5)[20.5, 0.118] 3 Phage display (subtractive panning) 3 PMID:25908560 Anti-thiacloprid monoclonal antibody 3A5 Thiacloprid Not determined. Not determined. X8 M13 phage display library Competition experiment The evaluations of the phage-displayed peptides were based on the IC50 (ug/L) and Amax/IC50 (the ratio of Amax to IC50), the combination of low IC50 and high Amax/IC50 was the most desirable. To eliminate nonspecific binding of phage to BSA, phages were firstly incubated with plate coated with BSA-PBS. After incubation, the supernatant containing unbound phages was incubated with the the plate containing 3A5 mAb. To elute the bound phage using competitive elution, 100 μL of thiacloprid (10 mg/L in PBS) was added to each well with shaking for 1 h to compete the binding phage from the coating antibody. 2870 VRFNDL[1.89] TRLNDL[1.93] TRIQDIN[1.94] TRINDV[1.95] TRVQDLPNW[2.06] TRMNDLKS[1.93] ERINDLTD[2.1] LRLNDLNTY[2.13] RYNDIEQH[1.9] LRINDLT[1.86] RWNDISE[1.89] SRVYDIL[1.94] NRINDFSD[2.02] T-R-I-N-D-L-T 13 Phage display (common panning) 3 PMID:25936577 Anti-GapC (1-150) mAb 1F2 Glyceraldehyde-3-phosphate dehydrogenase (EC:1.2.1.12) Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of 13 positive phage clones to the mAb1F2 were measured by sandwich ELISA. The OD values at 405 nm were measured and data shown were reproduced from the Fig. 3 in the reference. Wild-type M13 phage was used as a negative control, which had the value of 0.21 when binding to the mAb1F2. Amino acid sequence of the motif exactly matched 30TRINDLT36 of the S. dysgalactia GapC. Subsequently, site-directed mutagenic analysis further demonstrated that residues R31, I32, N33, D34 and L35 formed the core of 30TRINDLT36, and this core motif was the minimal determinant of the B-cell epitope recognized by the mAb1F2. The epitope 30TRINDLT36 showed high homology among different streptococcus species. Overall, our findings characterized a conserved B-cell epitope, which will be useful for the further study of epitope-based vaccines. 2871 AWTCSSRLCELG[0.2 ± 0.02] AQRYCTQSVCVW[0.56 ± 0.01] VCNEAVCYIESW[0.51 ± 0.02] AVCDALWCAFTA[0.88 ± 0.02] VGTCDDRWCVIR[1.28 ± 0.01] AYNCLHPTLCLI[1.22 ± 0.02] VCNSLWCTLPVG[0.86 ± 0.02] QNMFCSIATKSC[0.77 ± 0.02] C-D-x(2)-W-C 8 Phage display (common panning) 4 PMID:25944706 His-tagged porcine reproductive and respiratory syndrome virus (PRRSV) nonstructuralprotein (nsp) 7 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of phage clones encoding peptides with high binding affinities to His-tagged nsp7 proteins were identified by ELISA. The OD values at 450 nm were measured and data shown were reproduced from the Fig. 2 in the reference. The experiments were performed in triplicate and presented as mean ± standard deviation. Uncorrelated phage was served as the control, which had the value of 0.06 ± 0.02 when binding to the hHMGR protein. 2872 AQRYCTQSVCVW[0.56 ± 0.01] AVCDALWCAFTA[0.88 ± 0.02] VGTCDDRWCVIR[1.28 ± 0.01] AYNCLHPTLCLI[1.22 ± 0.02] VCNSLWCTLPVG[0.86 ± 0.02] QNMFCSIATKSC[0.77 ± 0.02] C-D-x(2)-W-C 6 Phage display (common panning) 5 PMID:25944706 His-tagged porcine reproductive and respiratory syndrome virus (PRRSV) nonstructuralprotein (nsp) 7 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of phage clones encoding peptides with high binding affinities to His-tagged nsp7 proteins were identified by ELISA. The OD values at 450 nm were measured and data shown were reproduced from the Fig. 2 in the reference. The experiments were performed in triplicate and presented as mean ± standard deviation. Uncorrelated phage was served as the control, which had the value of 0.06 ± 0.02 when binding to the hHMGR protein. 2873 VCNEAVCYIESW[0.51 ± 0.02] AVCDALWCAFTA[0.88 ± 0.02] VGTCDDRWCVIR[1.28 ± 0.01] AYNCLHPTLCLI[1.22 ± 0.02] VCNSLWCTLPVG[0.86 ± 0.02] C-D-x(2)-W-C 5 Phage display (common panning) 6 PMID:25944706 His-tagged porcine reproductive and respiratory syndrome virus (PRRSV) nonstructuralprotein (nsp) 7 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of phage clones encoding peptides with high binding affinities to His-tagged nsp7 proteins were identified by ELISA. The OD values at 450 nm were measured and data shown were reproduced from the Fig. 2 in the reference. The experiments were performed in triplicate and presented as mean ± standard deviation. Uncorrelated phage was served as the control, which had the value of 0.06 ± 0.02 when binding to the hHMGR protein. 2874 SGVYKVAYDWQH(16)[2.77 ± 0.29] GFPTRFEALSSN(2)[1.57 ± 0.25] VHWDFRQWWQPS(2)[1.52 ± 0.1] HTSSLWHLFRST(2)[2.25 ± 0.2] GLHTSATNLYLH(1)[2.23 ± 0.41] HSFKWLDSPRLR(1)[2.57 ± 0.12] SGVYKVAYDGQH(1)[2.3 ± 0.18] KASGSPSGFWPS(1)[3.1 ± 0.59] RRVDKVQYDRQH(1)[1.51 ± 0.12] GLHTSALSDLH(1)[NT] 10 Phage display (subtractive panning) 4, 5 PMID:25955351 Cation-independent mannose-6-phosphate receptor Insulin-like growth factor II 2L29 ,2V5P , Not determined. Ph.D.-12 phage display library (X12) ELISA Binding of the phages to IGF2R and BSA was examined with phage ELISA. The binding ratio OD450(IGF2R)/OD450(BSA) was calculated and data shown were reproduced from the Figure 3B in the reference. The phage with no insert peptide was served as the control, which had the value of 0.89 ± 0.11. The experiments were performed in triplicate and presented as mean ± standard deviation. NT denotes not tested. The first, third, and fifth rounds of biopanning were conducted on recombinant human IGF2R protein, while the second and fourth rounds of biopanning were conducted against rat hepatic stellate cells HSC-T6. Among the identified peptide candidates, peptide VHWDFRQWWQPS shows the highest binding affinity and specificity to recombinant human IGF2R protein and HSCs. The equilibrium dissociation constant (Kd) of peptide-431 is 6.19 μM for LX-2 cells and 12.35 μM for rat hepatic stellate cells HSC-T6. Cellular uptake of peptide-431 in LX-2 cells is significantly reduced after silencing IGF2R with siRNA. Peptide VHWDFRQWWQPS also enhances the uptake of a proapoptotic peptide (KLA peptide) in LX-2 and HSC-T6 cells, indicating that peptide VHWDFRQWWQPS can be used as a targeting ligand to deliver antifibrotic agents into not only rat but also human HSCs. Dimerization of peptide VHWDFRQWWQPS further increase its binding affinity to LX-2 cells by approximately 9-fold. 2875 YLDAWENEVINF[0.77 ± 0.06] YGDRWFMPVMRS[1.38 ± 0.11] DGYNGEPWIWQQ[0.77 ± 0.04] YAGDFWDLDALA[1.55 ± 0.004] WGYDWWENRSLT[0.11 ± 0.1] 5 Phage display (common panning) 3 PMID:25998380 Anti-TIM polyclonal antibody (pAb) Corticoliberin 3EHT ,3EHU , Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of positive phage clones to EC187 were measured by sandwich ELISA. The OD values at 450 nm were read. The experiments were presented as mean ± standard deviation. After 3-round biopanning, positive clones were selected and the polypeptides carried by them were identified. 5 polypeptides were found to bind with the target specifically. Among them, the peptide YAGDFWDLDALA exhibited the highest affinity. By evaluating the cAMP accumulation in the CRFR1 or CRFR2-expressing HEK293 cells, we demonstrated that peptide YAGDFWDLDALA blocking the function of CRFR1, but not CRFR2. In addition, we also found that P7 and CRF act on CRFR1 competitively. 2876 YHTTDKLFYMMR[1.78 ± 0.08] YSAYEFEYILSS[1.71 ± 0.05] KTMSAEEFDNWL[1.95 ± 0.11] LTSHTYRSQADT[0.68 ± 0.12] 4 Phage display (common panning) 4 PMID:26033236 Tocilizumab Interleukin-6 receptor Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Binding capabilities of the selected phages toward Tocilizumab was estimated by ELISA. The absorbance at 405 nm was measured and data shown were reproduced from the Fig. 1C in the reference. The experiments were performed in triplicate and presented as mean ± standard deviation. The mimotopes were conjugated to immunogenic carrier proteins and used to intraperitoneally immunize BALB/c mice. Sera from the mimotopes immunized mice not only showed specific binding to recombinant IL-6R, but can also IL-6R expressed in Hela, U-937, Jurkat cell lines and in fibroblast-like synoviocytes from patients with RA (FLS-RA). 2877 NKELQTV(7)[1.56 ± 0.2, 0.68 ± 0.18] VKELDTR(3)[1.78 ± 0.32, 1.22 ± 0.16] AKELSTW(2)[2.25 ± 0.23, 0.79 ± 0.32] K-E-L-x-T 3 Phage display (subtractive panning) 4 PMID:26048649 Imidacloprid hapten IMI Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding activities of positive phage clones to IMI-BSA were measured by competitive phage ELISA respectively using 0 and 1000 ng/mL of imidacloprid. The absorbance at 450 nm was measured and data shown were reproduced from the Figure 2 in the reference. The conjugate IMI-BSA was used as binding-templates to select peptides specific for imidacloprid. Bound phages in the wells were eluted. The eluted phage solutions were transferred onto wells coated with 3% BSA alone to eliminate the non-specific binding phages. 2878 PHSNRKKKDTQR[23.95 ± 0.38] HYKYYPTASVMK[104.1 ± 0.76] FTKVKPKSHNMT[23.95 ± 3.02] WPLSTVASVRVP[39.52 ± 2.67] VPRSMAATHSTF[14.83 ± 2.64] PHSNRKKNQYLE[23.19 ± 5.3] QTWDLPRTTHKT[11.79 ± 2.28] 7 Phage display (competitive panning) 3 PMID:26052070 Site-specifically biotinylated Complement C3 beta chain (C3b) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR) Direct binding of peptides was measured using SPR by injecting 1 mM of each peptide over site-specifically immobilized C3b. Data shown were reproduced from the Fig. 4B in the reference. A control peptide (YHPNGMNPYTKA) exhibits minimal response at this concentration, which had the Resonance Units (RU) value of 3.81 ± 0.76. Following washes of increasing stringency, bound phage were eluted using 100 μM MSCIN-A to specifically select for SCIN-competitive, C3b-binding sequences. Seven unique sequences exhibited direct C3b binding. A subset of these specifically inhibited the AP inassays of complement function. The mechanism of AP inhibition by these peptides was probed through surface plasmon resonance approaches, which revealed that six of the seven peptides disrupted C3bBb formation by interfering with factor B/C3b binding. 2879 KVWMPNNRGPVP AYTTSIPNRNQL TILKAWPNITQL AWPPPSMGPLAY KISQPTTVASLQ ILANDLTAPGPR TPFPFAPLGRPP KVWISPQSLGAT VSRHQSWHPHDL KCCTLPRPLEML NQRVLPPSHSWL LSLNFDRESETN KVWPPMYLSHTF AVPHRVGGLHSL NADNQMTWRHVL IDAPANQRLLQK KIWQMDQDSVRN HPHHETSVMRLQ KVWXMAPTTAFS SHPWNAQRELSV KVWQLQPSNSVT MKVSEPLHAHFS KAWVLTEWQTTK SIHRHHDDHFLT LLXKTLNNRLYD KVVQLSELSRLL DPALRHTHHNLR AHSHKLFLLSNR HSPSNLGFQSPP WMADTSPSLAST SWNSKAWIVVPA STMNDIHMELHR KCCFTTASSTSI GLKIWSLPPHHG KIXIFYPFKSPLI FPAMSFAXKKLA HFKPMQPAHPIP NSTLKVLPQGWM KVWPPLTSIVPS NLRTDSLTLVIR 40 Phage display (common panning) 3 PMID:26071378 Type IV secretion system protein virB10 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2880 DAQWAFM PPLSAPN GETPNPH LSSPTSN PLWDKRH NSWPPSR RLPHSVG QWTNHAL SGTPWHP DPFHVHN TTNSPPL FSSSRNY PATYTSA TQPWTRA NDFKLQF YKLPSRH ALGHSAK QKAHHEQ HEARSTM VIPQPQQ SYPTRPT FGNPHGH MRNDGAN PVPYPQL PKKLDSI KATAQGA HNPLLSF KPASHNA DSLQYPA TLPLPNL TNPVTES MSKHQAY PEAALSS TFLDTRA DPGASHP MERPAHL STHTSRL WAAIFPL PSKLQEH GPKMFRI LKITGED NRYTPTA LSYLHPT HPQLPSI NRGQLST GFLGKNQ KSAKTYH APSPWKQ PHTLSRT FSNQTML PAHRVPS CSTPTPH GMMPVAL 53 Phage display (common panning) 3 PMID:26071378 Type IV secretion system protein virB10 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2881 TQYYAYSTTQKS TQAIRVHTISSQ GLNQVLRIPSFI SPKHNLDMVKMM MIVDHLPIQVNT EYDYACGVVGYE SYPKASLALLAP 7 Phage display (common panning) 3 PMID:26058474 Magnesium fluoride (MgF2) particles Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2882 NQTHDWVPYAGR(8) YWPDERPFAARK(4) LHNPWRDTPPFA(4) TALDFPTWTAKN(4) VIDIPPFAYRNT(3) YADTPPFASTHK(2) STSDFLPYAARF(2) LPGVRDTPPFAA(2) VIGDMPPYAAMH(2) QWDLPPYAGRYD(2) GIDQLPWAARNK(2) YGIPWLDMPIKR(2) NQTDGQDRVPYA(1) TSPDAYPYAARQ(1) SDLPPFAAGRYN(1) NPFDTVPYAGRQ(1) RTLDWYDRVPYA(1) IQDWPPYASSTK(1) GERPPDYPPYAA(1) NHNQDFRPFPDR(1) KWQSDWPPFAAY(1) SDVDRPPWPART(1) SYVPDWPPFASK(1) APNVTDVPPFAA(1) IDTPPYAGRSSL(1) TLLIQRSPYEQI(1) SINPNPPLRPSY(1) D-x-[PVLP]-[YFW]-A-[AGS]-[RK] 27 Phage display (subtractive panning) 5 PMID:25520269 Anti-coagulation factor VIII monoclonal antibody (mAb) 2-76 Coagulation factor VIII Not determined. Not determined. Ph.D.-12 phage display library (X12) For selection of peptides that bind to the murine monoclonal anti-FVIII antibody (mAb) 2-76 immobilisation on streptavidin-coated beads was achieved via a biotin-conjugated rabbit anti-mouse IgG. For negative selections, an appropriate IgG isotype (murine IgG2a) was used. Three positive and two negative selections were performed. 2883 NPPKWPR(1) 1 Phage display (subtractive panning) 5 PMID:25520269 Anti-coagulation factor VIII monoclonal antibody (mAb) 2-76 Coagulation factor VIII Not determined. Not determined. Ph.D.-7 phage display library (X7) For selection of peptides that bind to the murine monoclonal anti-FVIII antibody (mAb) 2-76 immobilisation on streptavidin-coated beads was achieved via a biotin-conjugated rabbit anti-mouse IgG. For negative selections, an appropriate IgG isotype (murine IgG2a) was used. Three positive and two negative selections were performed. 2884 NFTHPGV(2) SSTVYGR(2) SKLDGEK(1) MHMYSPP(1) YSTEYTY(1) STHPLPP(1) NGTRYLG(1) WSRPPIM(1) LTLPRSF(1) FSAFPTS(1) LTSPLHI(1) RLPPQTP(1) WSTSYTM(1) DPKNQPA(1) LSAIPTR(1) 15 Phage display (subtractive panning) 5 PMID:25520269 Anti-coagulation factor VIII polyclonal antibody IgG from haemophilia A patient 1 Coagulation factor VIII Not determined. Not determined. Ph.D.-7 phage display library (X7) For positive selections, IgGs from FVIII inhibitor-positive plasma was immobilized on streptavidin-coated beads via a biotin-conjugated goat anti-human IgG. For negative selections, IgGs from a plasma pool of healthy individuals negative for FVIII-specific antibodies were immobilised likewise. 2885 NYTHPGS(1) WTHPGNR(1) NFTHPGS(1) WTHPGAQ(1) SFTHPGQ(1) YTHPGAA(1) T-H-P-G 6 Phage display (subtractive panning) 5 PMID:25520269 Anti-coagulation factor VIII polyclonal antibody IgG from haemophilia A patient 1 Coagulation factor VIII Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) For positive selections, IgGs from FVIII inhibitor-positive plasma was immobilized on streptavidin-coated beads via a biotin-conjugated goat anti-human IgG. For negative selections, IgGs from a plasma pool of healthy individuals negative for FVIII-specific antibodies were immobilised likewise. 2886 SNDFIHTWSTNW(3) DGNYTHTRSTNY(2) TPPYLHHFSTRY(2) QKSLHTFSTHYS(1) NCWTHPGATQCN(1) QGYNHVISTAYD(1) LTPITQLAGPMP(1) AIPYNHSVATYY(1) MGAHYRPQPNPH(1) LTYPDFLHNNFP(1) MYIPNNTPSMMR(1) LQVPWYATSTAK(1) FITPPSLPAFTG(1) ETQIHMVSTRYD(1) SMNYYHTFATNY(1) SPWLHTTSTHYL(1) STASNFTHPGND(1) SFTHRESTIYNA(1) DMIHTHYSSTRY(1) 19 Phage display (subtractive panning) 5 PMID:25520269 Anti-coagulation factor VIII polyclonal antibody IgG from haemophilia A patient 1 Coagulation factor VIII Not determined. Not determined. Ph.D.-12 phage display library (X12) For positive selections, IgGs from FVIII inhibitor-positive plasma was immobilized on streptavidin-coated beads via a biotin-conjugated goat anti-human IgG. For negative selections, IgGs from a plasma pool of healthy individuals negative for FVIII-specific antibodies were immobilised likewise. 2887 AMRVEPHTYKNH(7) AMLVDPQGAHSW(1) NPVEWFMSTVNT(1) GMKVEPWMPPPR(1) YMPVEPQSTYTK(1) AMPVESELYKLM(1) AMPVAPDTQRYR(1) SFNTRPLPFLSY(1) AQRVDPDLVSIS(1) [AG]-M-x-V-[ED]-P 9 Phage display (subtractive panning) 5 PMID:25520269 Anti-coagulation factor VIII polyclonal antibody IgG from haemophilia A patient 3 Coagulation factor VIII Not determined. Not determined. Ph.D.-12 phage display library (X12) For positive selections, IgGs from FVIII inhibitor-positive plasma was immobilized on streptavidin-coated beads via a biotin-conjugated goat anti-human IgG. For negative selections, IgGs from a plasma pool of healthy individuals negative for FVIII-specific antibodies were immobilised likewise. 2888 AMLVQPA(5) AMPVEPD(2) AMFVEPA(1) AMLVFPE(1) AMQVDPA(1) AMPVQPH(1) AMLVEPY(1) LTLQPTF(1) [AL]-M-x-[VQ]-E-P 8 Phage display (subtractive panning) 5 PMID:25520269 Anti-coagulation factor VIII polyclonal antibody IgG from haemophilia A patient 3 Coagulation factor VIII Not determined. Not determined. Ph.D.-7 phage display library (X7) For positive selections, IgGs from FVIII inhibitor-positive plasma was immobilized on streptavidin-coated beads via a biotin-conjugated goat anti-human IgG. For negative selections, IgGs from a plasma pool of healthy individuals negative for FVIII-specific antibodies were immobilised likewise. 2889 SLQPTEHNKFWL(2) MTTPTESHKFGH(1) DLLRPSEKNKFF(1) SYTPSELRVFQT(1) TPAPSERMHFDS(1) SPSELQKFQTRS(1) KSTPSEIRHFDY(1) [TS]-P-[ST]-E-x(2)-[KH]-F 7 Phage display (subtractive panning) 5 PMID:25520269 Anti-coagulation factor VIII polyclonal antibody IgG from haemophilia A patient 5 Coagulation factor VIII Not determined. Not determined. Ph.D.-12 phage display library (X12) For positive selections, IgGs from FVIII inhibitor-positive plasma was immobilized on streptavidin-coated beads via a biotin-conjugated goat anti-human IgG. For negative selections, IgGs from a plasma pool of healthy individuals negative for FVIII-specific antibodies were immobilised likewise. 2890 DAWIGSRIPFKG(14) LCLDISCHIPTK(2) SHSAIQDHMPRK(1) RTWHPSEIMHEY(1) VHMKQIPSWLIK(1) LDKQTLKYLHEK(1) VEGIAGHIPTKL(1) I-x(2)-[HR]-[ILM]-P-x-K 7 Phage display (subtractive panning) 5 PMID:25520269 Anti-coagulation factor VIII polyclonal antibody IgG from haemophilia A patient 10 Coagulation factor VIII Not determined. Not determined. Ph.D.-12 phage display library (X12) For positive selections, IgGs from FVIII inhibitor-positive plasma was immobilized on streptavidin-coated beads via a biotin-conjugated goat anti-human IgG. For negative selections, IgGs from a plasma pool of healthy individuals negative for FVIII-specific antibodies were immobilised likewise. 2891 GIIIPHQ[0.95 ± 0.04] MIPHDEN[1.16 ± 0.04] VHALVPH[0.85 ± 0.05] FSLIPHS[1.01 ± 0.09] ISLIPHT[1.03 ± 0.08] FVPHANK[0.95 ± 0.04] IMPLWPH[0.84 ± 0.04] LIPHDSL[1.19 ± 0.02] LNPNKLI[0.81 ± 0.05] WNLVPHT[0.88 ± 0.04] [IV]-P-H-D 10 Phage display (common panning) 3 PMID:25741007 Anti-avian hepatitis E virus (HEV) capsid protein monoclonal antibody 3E8 Capsid protein Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Affinities of phages to mAb 3E8 were measured by phage ELISA. The OD values at 490 nm were measured and data shown were reproduced from the Fig. 2A in the reference. Error bars indicate the standard range of individual values from two independent experiments and the results were presented as mean ± standard deviation. 2892 KLAPHAN[1.07 ± 0.13] GLKLYQS[1.68 ± 0.06] FKLYMEP[1.63 ± 0.04] VKLYMTF[1.81 ± 0.04] VHALYLQN[0.96 ± 0.02] LDPARF[1.09 ± 0.15] GNLFMTL[1.43 ± 0.07] VKLFMSTL[1.73 ± 0.05] RDAVRPF[1.25 ± 0.03] V-K-L-Y-[MT]-S 9 Phage display (common panning) 3 PMID:25741007 Anti-avian hepatitis E virus (HEV) capsid protein monoclonal antibody 1B5 Capsid protein Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Affinities of phages to mAb 1B5 were measured by phage ELISA. The OD values at 490 nm were measured and data shown were reproduced from the Fig. 2B in the reference. Error bars indicate the standard range of individual values from two independent experiments and the results were presented as mean ± standard deviation. 2893 RATQLPQ(4/12) QHIPKPP(2/12) HRRPSRS(1/12) AFDTHTM(1/12) TPPEPAP(1/12) NQDVPLF(1/12) HRLRISP(1/12) GDTQRVA(1/12) 8 Phage display (common panning) 4 PMID:25947144 δ2 CDR3 peptide CACDVLGVNTDKLIFGKG Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Four peptides that bound to the predominant δ2 CDR3 fragments and showed homology to M.tb genes in a BLAST search. Notably, one fragment was related to M.tb H37Rv (QHIPKPP), and this fragment was confirmed as a ligand for the γδ TCR. 2894 SEISAST(2/10) ATKTRQP(2/10) LNPVHRL(1/10) YPTGLPL(1/10) RFRPTPP(1/10) ADHPPPH(1/10) SRVRLGA(1/10) HRSHSTH(1/10) 8 Phage display (common panning) 4 PMID:25947144 δ2 CDR3 peptide CACDTLGDRDYTDKLIFGKG Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2895 GWHHHPR(5/12) HKRPRNN(1/12) HRRPSRS(1/12) RRRPMAI(1/12) HGSTKRT(1/12) WNPHKHH(1/12) WRQTRKD(1/12) GTTTTLL(1/12) 8 Phage display (common panning) 4 PMID:25947144 δ2 CDR3 peptide CACDTIGTGGHYTDKLIFGKG Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2896 SPRVGAT(2/13) ATKTRQP(1/13) HKRPRNN(1/13) AFDTHTM(1/13) SEISAST(1/13) QHIPKPP(1/13) RRAPTPP(1/13) YMQLPSH(1/13) KLAPMTS(1/13) GWHHHPR(1/13) ASPDTPQ(1/13) 11 Phage display (common panning) 4 PMID:25947144 δ2 CDR3 peptide CACDTLWGIQGNTDKLIFGKG Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2897 TDFNTMAKNNPP[4.97] 1 Phage display (common panning) 3 PMID:26040578 Biotinylated anti-lysozyme polyclonal antibody IgG Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Binding capabilities of the selected phage toward biotinylated anti-lysozyme polyclonal antibody IgG was estimated by ELISA. The absorbances at 418 nm were measured and data shown were reproduced from the Fig. 4 in the reference. The affinity value was 4.97 when the biotinylated anti-lysozyme polyclonal antibody IgG was at a concentration of 3e-8. 2898 STGTPRA(3) KSTSQPE(2) TRTPQHS(1) LSTRAPL(1) DRAPGRT(1) LSPARTT(1) PPYLSTR(1) TTTPTLH(1) TMTGSTT(1) PYSAKAH(1) PPYLSHL(1) RPAPNQT(1) PHPISKQ(1) TNTAWTS(1) MTSSGML(1) HHTTSTG(1) ISQPIRQ(1) 17 Phage display (common panning) 4 PMID:25802296 Biopolymer transport protein ExbD (43–141) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2899 FSISTLQSAPTR(2) QTTAWWGAPARL(2) QMMQTSSSPPTV(1) STNPAALYSDYS(1) HHRAHNSHLSVR(1) HSIFYPIYLPSQ(1) FHESWPSPAGGR(1) FPHYPVSTLYSL(1) STPQPSLFSYPV(1) FHSSTPTAPPQK(1) NGLTSSRPWSFL(1) SPIYVTWVPTAL(1) NDPGRLRVPVST(1) RHYEPLSRVSSS(1) STPQVYNVFYAP(1) FHSHWPSMADNS(1) TVYWITPPALPI(1) FHETWPARVSYL(1) SHHWEPISSPLR(1) KIYPITLTYLAP(1) SSKVLPSSFFTR(1) APTTQTPPINWK(1) QTNSQHPISALR(1) RHSEPISVFYIT(1) HPIYVTYYPDPS(1) HLLMKPPQTSPA(1) 26 Phage display (common panning) 3 PMID:25802296 Biopolymer transport protein ExbD (43–141) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2900 SHSNTTQTRPSD SHALPLTWSTAA NTIPMHTSTHTI IHPASQSRQNTT AALGTYSTHTPT HLPTSSLFDTTH HGLPVTTRGAFG TKTVAQTTTSIS KLVDESSTSPLS HSNLPTKRPTSL 10 Phage display (common panning) 3 PMID:25802296 Protein TonB (33-239) Ferrienterobactin receptor Not determined. Not determined. Ph.D.-12 phage display library (X12) 2901 NSTSSPQ LPASWHP SMNTFQP NPTPEKR KTSLPRL RTFDLPA PQPKTYQ SHLPAAL LTQTPTR 9 Phage display (common panning) 4 PMID:25802296 Protein TonB (33-239) Ferrienterobactin receptor Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2902 LPLSRHY(3) NTHHTSM(3) HLRHLTH(3) HTWRHHS(2) ANITHHH(2) HLRPHIH(2) HLNAHKH(1) QTDHSHS(1) HKHIPSH(1) HSSHHNH(1) HTRDHHH(1) HVHPSNA(1) HSGPMIR(1) LPTPPAP(1) RASPPAT(1) HEWTTHA(1) HAPTAHL(1) LTQSHTG(1) 18 Phage display (common panning) 3 PMID:25930968 Membrane protein (FXYD2)γa Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2903 LPLSRHY(3) NTHHTSM(3) HDRLKSH(1) IHAHLPQ(1) QSNLHTI(1) HRFHHQE(1) HTNHQLH(1) PLQSSPR(1) SMLPHSP(1) LGRVQMM(1) NHAHSTP(1) DPAPRPR(1) DPWPWVK(1) LAVRSPP(1) HTRHIPH(1) 15 Phage display (common panning) 4 PMID:25930968 Membrane protein (FXYD2)γa Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2904 HSMSLLTSHYHL(2)[2.07 ± 0.04] HYGFFSTHPLQH(1)[0.48 ± 0.06] HQPHHRYGSPFN(1)[0.63 ± 0.04] HNYPQYPRPPDV(1)[0.18 ± 0.01] WTKHHNHVHTPP(1)[0.91 ± 0.03] DISKMYLGPPPY(1)[0.63 ± 0.06] TLNMHLFPFHTG(1)[1.4 ± 0.04] AAMPLHWPGITQ(1)[0.65 ± 0.04] FDFPFWLRNPAP(1)[0.46 ± 0.03] 9 Phage display (common panning) 4 PMID:26013256 Recombinant S-AD (rS-AD) protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of the selected phage clones to the recombinant S-AD (rS-AD) protein were measured by ELISA. The OD values at 490 nm was measured and data shown were reproduced from the Fig. 1 in the reference. The experiments were performed in triplicate and presented as mean ± standard deviation. The non-binding phage library was served as the control, which had the value of 0.11 ± 0.03 when binding to the recombinant protein. Among the 9 phages selected that specifically bound to rSAD, the phage bearing the peptide TLNMHLFPFHTG bound with the highest affinity and was subsequently used to develop a phage-based ELISA for TGEV. 2905 NCEDYLPGWFCT(14/29) DCNARQAYSRCA(1/29) GCDWYAIWRTCI(1/29) HCLSRRAVLFCA(1/29) GCPPVVWRPSCS(1/29) TCLPDLQHSTCY(1/29) GCPRTTHDQACA(1/29) FCATPHTRPLCH(1/29) RCVPSREDPWCL(1/29) QCYKTEARITCY(1/29) SCERQYTPSECQ(1/29) KCRRGFLAHSCQ(1/29) 12 Phage display (common panning) 3 PMID:25944054 Plasmodium berghei female gametes Not determined. Not determined. Not determined. LX-8 phage display library (XCX8CX) After three rounds of selection, close to half of the recovered phages displayed the same NCEDYLPGWFCT peptide, named FG1 for Female Gamete peptide 1, which binds specifically to Plasmodium berghei female gametes. FG1 inhibits gamete fertilization and interferes with further development of the parasite in the mosquito. 2906 WHPWSYLWTQQA(12)[0.78 ± 0.05] TLWHPWHYPAMR(8)[0.62 ± 0.03] EYVSPYAGPTHQ(3)[0.52 ± 0.04] NNGHAQIYMVHK(3)[0.41 ± 0.02] WHYNSWYRWPVM(2)[0.39 ± 0.02] SYHHTPHFAGPP(1)[0.31 ± 0.02] HWWSWYTPFNHT(1)[0.28 ± 0.04] 7 Phage display (subtractive panning) 4 PMID:26140900 CD44 antigen protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Affinities of the positive phage clones to CD44 protein were measured by phage ELISA. The A450 values were measured and data shown were reproduced from the Fig. 1b in the reference. Unrelated phage were used as negative controls, which had the value of 0.20 ± 0.02 when the phage binded to the target. Phages from the library were firstly incubated with BSA. After incubation, the unbound phages was incubated with CD44 protein. The WHPWSYLWTQQA peptide exhibits affinity and specificity to CD44 on cells and has the potential to be used as a candidate probe for gastric cancer cell targeting. 2907 GWYEHNNYYQGV[1.24] ALYTLYPPDSLH[1.30] DRALYGPTVIDH[1.52] ALWPPNLHAWVP[1.45] 4 Phage display (common panning) 3 PMID:26267086 Anti-bovine herpesvirus type 1(BoHV-1) polyclonal antibody IgG Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Affinities of the positive phage clones to purified IgG, by affinity chromatography, from a serum bank of seropositive bovines infected with BoHV-1 were measured by phage ELISA. The OD values were measured at 490 nm and data shown were reproduced from the Fig. 1a in the reference. The wild-type phage M13KE (Wt) was used as a negative control, which had the value of 0.52 when binding to the target. 2908 IDNSHTH(5) SVTIHHL(1) ITLREHH(1) KMISATE(1) VKLNLSG(1) GAMSHHK(1) ETSNTRL(1) HAGFVPS(1) YHAEHFN(1) LDHKWHK(1) GLSHEIH(1) HGGVRLY(1) HAGFVPS(1) SQLPFHH(1) HLNQQNH(1) KVLHGPH(1) 16 Phage display (subtractive panning) 3 PMID:26177065 Agrobacterium vitis polygalacturonase Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Affinities of the positive phage clones to agrobacterium vitis polygalacturonase (PG) were measured by phage ELISA. The A490 values were measured and data shown were reproduced from the Fig. 2 in the reference. Data are represented as the mean of three replicates, and error bars represent the 95% confidence interval. In the first round of panning, phages were incubated with agarose beads bound with A. vitis polygalacturonase. Beads were harvested by low-speed centrifugation for 30 s, and the supernatant containing unbound phage was removed. The agarose beads were then washed 10 times with PBST and phages were eluted. In the second round, phages were incubated with blocked agarose beads containing no bound polygalacturonase to eliminate target-unspecific phage from the phage pool. The supernatant from this step was used in a second round of panning with polygalacturonase-bound beads. The peptide SVTIHHLGGGS was able to reduce A. vitis polygalacturonase activity by 35% in vitro. Truncation studies showed that the IHHL motif alone is sufficient to inhibit A. vitis polygalacturonase activity. 2909 CNRWHHLEC(2)[0.87 ± 0.03] CTAHHTKMC(2)[NT] CGHRDMLHC(2)[NT] CNFLYSWTC(1)[0.69 ± 0.02] CDLVRGHHC(1)[NT] CNHHRLPSC(1)[NT] CYSNSFSLC(1)[NT] CPTNQHHLC(1)[NT] CLEAHDHKC(1)[NT] CIEYHGHSC(1)[NT] CPRWHHLGC(1)[NT] 11 Phage display (subtractive panning) 3 PMID:26177065 Agrobacterium vitis polygalacturonase Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Affinities of the positive phage clones to agrobacterium vitis polygalacturonase (PG) were measured by phage ELISA. The A490 values were measured and data shown were reproduced from the Fig. 2 in the reference. Data are represented as the mean of three replicates, and error bars represent the 95% confidence interval. In the first round of panning, phages were incubated with agarose beads bound with A. vitis polygalacturonase. Beads were harvested by low-speed centrifugation for 30 s, and the supernatant containing unbound phage was removed. The agarose beads were then washed 10 times with PBST and phages were eluted. In the second round, phages were incubated with blocked agarose beads containing no bound polygalacturonase to eliminate target-unspecific phage from the phage pool. The supernatant from this step was used in a second round of panning with polygalacturonase-bound beads. 2910 NLSSSWI(3/48)[0.047 ± 0.025] PAERLRS(2/48)[0.039 ± 0.024] FLPASGL(1/48)[0.104 ± 0.023] PWPLPYL(1/48)[0.030 ± 0.004] WGLLDLT(1/48)[0.050 ± 0.005] RNLDGWS(1/48)[0.023 ± 0.008] TLPSNTH(1/48)[0.021 ± 0.015] MSAFPFL(1/48)[0.027 ± 0.010] SRLGQYI(1/48)[0.040 ± 0.007] PFGPLPP(1/48)[0.038 ± 0.028] TIASTLH(1/48)[0.018] PRAPADV(1/48)[0.047 ± 0.003] ESPLKRQ(1/48)[0.069 ± 0.016] 13 Phage display (competitive panning) 3 PMID:26312490 TGF-β1 receptors onto peripheral blood mononuclear cells (PBMC) surface Transforming growth factor beta-1, TGF-beta-1 Not determined. Not determined. Ph.D.-7 phage display library (X7) Affinities of the selected phage clones to TGF-β1 receptors onto peripheral blood mononuclear cells (PBMC) surface were measured by phage ELISA. The OD492 values were measured and data shown were reproduced from the Fig. 1 in the reference. The wild-type phage was used as a negative control, which had the value of 0.030 ± 0.004 when this phage binded to the target. Phages that were bound to TGF-β1 receptors onto peripheral blood mononuclear cells (PBMC) surface were competitively eluted with 10 ng/mL of recombinant TGF-β1 (Sigma-Aldrich). 2911 NGSNRDIVEVQR[1.50] NQIYNRDYTEPT[1.45] YNRDMLETDYVN[1.48] QNTWNRDSIEET[1.46] FPEVSVNRLVVE[1.45] HVNRLHVEGPVP[1.49] KMTLPMNRSHVE[1.46] SYVTGGNRYAVE[1.50] SSYLSNRLFTEA[1.45] SATTMSNRYYTE[1.49] QPYNRSYIDFMV[1.31] N-R-[DLSY]-x-[VTI]-E 11 Phage display (common panning) 3 PMID:26135599 Anti-dengue virus 2 monoclonal antibody DB21-6 Dengue virus 2, DENV2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Affinities of the selected phage clones to mAb DB21-6 were measured by phage ELISA. The OD was measured at 490 nm and data shown were reproduced from the Fig. 3b in the reference. 2912 LSNRLHVESLEL[1.36] NQTNRHFVEIVH[1.97] SGLDRNRQLVER[1.50] NRTLVELGYAML[1.28] VNRPWVETTTQG[1.55] IVPYSNRTVTET[1.64] NRVSNEPFWDIA[1.44] DYLNRSTNEPAL[1.35] SMPLSGRAVVEG[1.61] HTSLHSGRNSVE[1.36] SSPGVISRFLVE[1.50] DRYLVEYSSGRW[1.36] MPSGGRFLVEGA[1.44] [NG]-R-x(2)-[VN]-E 13 Phage display (common panning) 3 PMID:26135599 Anti-dengue virus 2 monoclonal antibody DB39-2 Dengue virus 2, DENV2 Not determined. Not determined. Ph.D.-12 phage display library (X12) Affinities of the selected phage clones to mAb DB39-2 were measured by phage ELISA. The OD was measured at 490 nm and data shown were reproduced from the Fig. 3b in the reference. 2913 GGARDAGKAEWW(26.7%)[0.98 ± 0.04] VDVDETNNQSYP(6.7%)[0.60 ± 0.01] EGEPDPRSIVGP(6.7%)[0.47 ± 0.02] 3 Phage display (common panning) 4 PMID:26248692 Dengue virus serotype 2(DENV-2) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Affinities of the selected phage clones to dengue virus serotype 2(DENV-2)were measured by phage ELISA. The OD410 values were measured and data shown were reproduced from the Fig. 1 in the reference. The wild-type phage was used as controls, which had the value of 0.21 ± 0.01 when the phage binded to the target. 2914 MPHPTKNFDLYV ALWPPNLHAWVP 2 Phage display (subtractive panning) 6 PMID:25699101 BG1 ovarian cancer cell Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage display library was incubated with ovarian cancer cells (BG1), and un-captured phages were removed by the washing buffer. After five positive selection panning rounds, the positively selected phages were collected and incubated with other cancer cells. Unbound phages were collected and cloned into E. coli cells. 2915 DCAIVYAYDPCL(15/32) GCQPHEYSDPCA(12/32) ACLTTRPQPHCR(4/32) 3 Phage display (common panning) 4 PMID:26216124 Kupffer cell Not determined. Not determined. Not determined. LX-8 phage display library (XCX8CX) The peptide DCAIVYAYDPCL strongly binds to the Kupffer cell surface and inhibits sporozoite Kupffer cell entry. Furthermore, the peptide also binds to CD68, a putative receptor for sporozoite invasion of Kupffer cells that acts as a gateway for malaria infection of the liver. 2916 SKVGYPT[0.36 ± 0.16] YKASLIT[1.00 ± 0.16] ALGGVAM[0.66 ± 0.35] QSPRSIS[0.77 ± 0.06] ANLWPDG[0.80 ± 0.05] GPLSHKG[0.19 ± 0.18] NNNWPWT[0.70 ± 0.09] IQMTDIA[0.44 ± 0.30] SRHLHEW[0.36 ± 0.02] APVTSMK[0.34 ± 0.07] ATNKITK[NT] 11 Phage display (subtractive panning) 11 PMID:26346061 Chromium(III) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Affinities of the selected phage clones to Cr(III) were measured by phage ELISA. The A405 values were measured and data shown were reproduced from the Fig. 2 in the reference. The library was used as controls, which had the value of 0.30 ± 0.03 when the phage library binded to the target. A complete biopanning procedure consists of prenegative screening against blank NTA resin (1 round), positive screening against Cr(III) (3 rounds), and postnegative screening against the other metal species (Iron ion : 1 round, Cadmium ion : 2 rounds, Copper ion : 2 rounds, Zinc ion : 1 round, Mercury ion: 1 round). 2917 QRHKPRE 1 Phage display (common panning) 4 PMID:26181290 Extracellular domains(ECD) of epidermal growth factor receptor(EGFR) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2918 TSISINPPRRPS[1.12 ± 0.09] SRPRPLIRNRRP[1.06 ± 0.01] MTIRRHRHRPKI[1.07 ± 0.02] SRRRIPRINRPQ[1.08 ± 0.09] KRRNTILINLPN[1.22 ± 0.03] TPIKKMIRRLPH[1.23 ± 0.02] LPTKRIIKRMRR[1.18 ± 0.19] MSLNLRMRPMRI[1.02 ± 0.06] KMTRRTHINQIS[1.03 ± 0.08] RSIPRIHINTTN[1.04 ± 0.04] 10 Phage display (subtractive panning) 3 PMID:26417591 IgG from Alzheimer’s disease (AD) patients Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Affinities of the selected phage clones to IgG from Alzheimer’s disease (AD) patients were measured by phage ELISA. The results were expressed as an arbitrary ELISA Index (EI) and calculated as follows: EI =Abs of serum sample/cut-off, where the cut-off was determined as the mean absorbance of the negative control sera plus two standard deviations. Values of EI > 1.0 were considered positive. For subtraction of nonspecific peptides, the M13 phage library was added to TBS-Tween 0.1%. After 30 minutes of incubation, magnetic separation was performed. The phage eluate was subtracted two more times prior to the positive selection, which was performed for 30 minutes against IgG-coupled beads of AD patients, completing one selection cycle. 2919 GWSLSTV[1.57 ± 0.05] WRDNAIA[1.69 ± 0.06] WNLNTTN[1.72 ± 0.05] WNLNHIF[1.53 ± 0.07] WSQTTVS[1.57 ± 0.04] WGENTIW[1.16 ± 0.03] FNLNTAM[1.43 ± 0.07] WNSNAIS[1.55 ± 0.09] AWNFNSQ[1.48 ± 0.05] WNFTARE[1.15 ± 0.02] WNQNTAS[1.73 ± 0.04] WTYTTFE[1.17 ± 0.03] WSQTTVS[1.37 ± 0.05] WNQNTVS[1.76 ± 0.07] WNSNAIS[1.30 ± 0.05] 15 Phage display (common panning) 3 PMID:26289074 Anti-avian influenza virus nonstructural protein 1 MAb D9 Avian influenza virus nonstructural protein 1 Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Affinities of the selected phage clones to anti-avian influenza virus nonstructural protein 1 MAb D9 were measured by phage ELISA. The OD490 values were measured and data shown were reproduced from the Fig. 4 in the reference. Error bars represent the standard deviation of the results from three replicate experiments. 2920 YSPFHKWFPSMH(6)[0.83 ± 0.12] VSWKTWFPNLAV(2)[0.81 ± 0.17] IPQVWRDWFKLP(1)[0.70] FPAWFTKLYPRT(1)[0.98] QINTAKWWKTHF(1)[0.77] DASKALRSSGMP(1)[0.82] 6 Phage display (common panning) 4 PMID:26405107 Cholera toxin B protein, CTB Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Affinities of the selected phage clones to cholera toxin B were measured by phage ELISA. The A492 values were measured and data shown were reproduced from the Fig. 1b in the reference. 2921 PHLATLF(8/10)[2.86 ± 0.46] 1 Phage display (in vivo) 3 PMID:26081347 Ovarian cancer A2780 cell Not determined. Not determined. Not determined. FliTrx M13 phage display library (X7) ELISA Affinities of the selected phage clones to ovarian cancer A2780 cell were measured by cell-based ELISA. The binding of each phage to the A2780 cells was compared with the original library locus coeruleus. The phage optical density ratio of >2.1 indicated that specific binding to the A2780 cells. In the initial step, a 7-mer library was injected into nude mice through a vein. After three rounds of screening, phages were enriched. 2922 RKVFHASTS RKIVHAQTP KKVFATSGS ARRVMFVTT RRVLSAMHP RRAISTTVS RRAINNLPM RRSTTHAAA YRKTFQPPT KRFTMPPHP RRYTHPNNS KKAHPGGSP GKRASQSTL IWTKRVAKT 14 Phage display (common panning) 3 PMID:26164011 Histiocytic lymphoma cell line U937 Not determined. Not determined. Not determined. pVIII-9aa phage display library (X9) 2923 LMQINPTYYQIM(8/10) WSFNPSTYTIAG(1/10) HDFVADMYQLAQ(1/10) 3 Phage display (subtractive panning) 5 PMID:26292945 Anti-immunodominant region of the PEDV Spike protein (S1) monoclonal antibody 5E12 Spike protein S1 region Not determined. Not determined. Ph.D.-12 phage display library (X12) During first to third rounds of panning, the purified MAb-5E12 was incubated with the crude phage library or with eluted phage. In order to remove non-specific, antibody-binding molecules, in the fourth round of biopanning, an unrelated MAb was incubated with the third round-eluted phage. The unbound phage was subjected to a fifth round of panning. 2924 AHSGMYP 1 Phage display (common panning) 3 PMID:26299762 Small cell lung cancer (SCLC) H446 cell Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The peptide AHSGMYP exhibited excellent antitumor efficacy and particularly effectively inhibited the liver metastases with better tolerance. Results of cellular uptake and biodistribution indicated that the excellent antitumor efficacy of AHSGMYP was attributed to both the increased accumulation of drug and cellular uptake. 2925 ACDSCSYAKLCQ(7/25) KCLTQQPTTMCD(1/25) CCHSTREVRMCK(1/25) LCCRFASRSSCC(1/25) HCRIPPSQTVCE(1/25) ACRKYLPTIPCP(2/25) PCQNRLPRYICS(1/25) PCAGQSLAKLCQ(1/25) 8 Phage display (common panning) 4 PMID:26341796 Caprine umbilical vein endothelial cell Not determined. Not determined. Not determined. LX-8 phage display library (XCX8CX) 2926 APLSPPG(6/30) APLSRPG(2/30) APLSRRG(1/30) APLPRPG(1/30) GPLSRPG(1/30) HPLMRRG(1/30) HPLIVLE(1/30) AVLRTSP(1/30) APSPMIW(1/30) HSLELMP(1/30) RLPAFLH(1/30) LMPARYH(1/30) HLWLIPP(1/30) TYASDRS(1/30) QYATPSA(1/30) QMAQWPP(1/30) SWNMWSP(1/30) HWGMWSY(1/30) VNSAWTY(1/30) TTTWHLK(1/30) NQSAVLP(1/30) DSHTPQR(1/30) GHWTRFA(1/30) GHWTRFA(1/30) 23 Phage display (common panning) 3 PMID:26369825 Amino acid sequence FDYSEPGNFSDISWPCNSSD from ACKR3/CXCR7 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2927 LLADTTHHRPWT(2/10) GQVLPTTTPSSP(1/10) MPFGARILSLPN(1/10) QLQLSNSMSSLS(1/10) SKHERYPQSPEM(1/10) HTHSSDGSLLGN(1/10) ANPIISVQTAMD(1/10) 7 Phage display (common panning) 3 PMID:26121597 Beta-tri calcium phosphate Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Beta-tri calcium phosphate powder was blocked for 24 hours at 4°C under moderate agitation with sterile filtered Odyssey Blocking Buffer (OBB; non-mammalian blocking buffer; LI-COR; Lincoln, NE, USA). Blocked Beta-tri calcium phosphate was pelleted at 2000 RPM for 2 minutes, washed 3X with PBS then subjected to three rounds of phage display. 2928 LLADTTHHRPWT(5/10) HNMAPATLHPLP(1/10) QSFASLTNPRVL(1/10) HTTPTTTYAAPP(1/10) QYGVVSHLTHTP(1/10) DYDSTHGAVFRL(1/10) 6 Phage display (common panning) 3 PMID:26121597 Beta-tri calcium phosphate Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Beta-tri calcium phosphate powder was blocked for 24 hours at 4°C under moderate agitation with 5% bovine serum albumin in phosphate buffered saline (BSA; Sigma-Aldrich, St. Louis, MO, USA). Blocked Beta-tri calcium phosphate was pelleted at 2000 RPM for 2 minutes, washed 3X with PBS then subjected to three rounds of phage display. 2929 LLADTTHHRPWT(1/10) VPQHPYPVPSHK(1/10) TMSNPITSLISV(1/10) IGRISTHAPLHP(1/10) MNDPSPWLRSPR(1/10) QSLGSMFQEGHR(1/10) KPLFTRYGDVAI(1/10) EPTKEYTTSYHR(1/10) DLNELYLRSLRA(1/10) NYDSMSEPRSHG(1/10) 10 Phage display (common panning) 3 PMID:26121597 The composite of beta-tri calcium phosphate and polylactide-co-glycolide (PLGA) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Beta-tri calcium phosphate powder combined with PLGA was blocked for 24 hours at 4°C under moderate agitation with 5% bovine serum albumin in phosphate buffered saline (BSA; Sigma-Aldrich, St. Louis, MO, USA). Blocked Beta-tri calcium phosphate was pelleted at 2000 RPM for 2 minutes, washed 3X with PBS then subjected to three rounds of phage display. 2930 APAIPAS(20/25) APALPAS(1/25) APSIPAS(1/25) 3 Phage display (common panning) 3 PMID:26172085 Anti-vibrio cholerae O1 Ogawa monoclonal antibody IXiao3G6 Vibrio cholerae serotype O1 Not determined. Not determined. Ph.D.-7 phage display library (X7) 2931 ALWPPQGHAWVP(7/10) HGENPPSGMAWV(1/10) HGVNPPSGMAWV(1/10) 3 Phage display (common panning) 3 PMID:26315775 Anti-S1 subunit of IBV Sczy3 monoclonal antibody 1D5 The spike (S) protein of the infectious bronchitis virus (IBV) Not determined. Not determined. Ph.D.-12 phage display library (X12) 2932 HIQTRLEPQWKS(8/10) HGIQTATEPQWS(1/10) HTLQTRTPPQSP(1/10) 3 Phage display (common panning) 3 PMID:26315775 Anti-S1 subunit of IBV Sczy3 monoclonal antibody 6A12 The spike (S) protein of the infectious bronchitis virus (IBV) Not determined. Not determined. Ph.D.-12 phage display library (X12) 2933 NKVIWEADWAFS(21/23) NKVIWDRDWMYP(1/23) HHSHSILQSDWF(1/23) 3 Phage display (common panning) 3 PMID:15344223 Anti-E. coli K1 monoclonal antibody HmenB3 Escherichia coli K1 bacteria Not determined. Not determined. Ph.D.-12 phage display library (X12) 2934 HKMHSHPRLTSP(6/35)[0.128] WHKAVPRWLASP(3/35)[0.543] FHKHKSPALSPV(2/35)[0.479] FHGHLKKPHWRN(2/35)[0.154] WHRHWPSHPTQK(1/35)[1.615] GLLHHKHHRSPY(1/35)[0.456] DLNYFTLSSKRE(1/35)[0.177] FHKHRISPSPST(1/35)[0.252] ATAKHLYWWRNQ(1/35)[0.656] WHKGNNVAWTKR(1/35)[0.430] WHSHMKTRTWQP(1/35)[0.543] FHSPKKNHHYYR(1/35)[0.507] HHKHSNRSPIFS(1/35)[0.224] FHKHHKSPRLFP(1/35)[0.268] AHKWYSQWLPHR(1/35)[1.461] WHHRHQPAPGGR(1/35)[1.021] FHKHSPRSPIFI(1/35)[0.371] AKLHWHKHHPLT(1/35)[<0.1] FHKHNYKSPPII(1/35)[<0.1] WHKVPRSDRMPP(1/35)[<0.1] FHKYSPPQKPVT(1/35)[<0.1] WPHFHRPPHREL(1/35)[<0.1] HLYHKNRNHIAY(1/35)[<0.1] IQRHHKPLRLRV(1/35)[<0.1] FHKHDRGRLSPP(1/35)[<0.1] SWRSRQLPETGE(1/35)[<0.1] 26 Phage display (common panning) 4 PMID:14748062 Anti-NS3 monoclonal antibody 3B1C4 Hepatitis C virus (HCV) nonstructural 3 (NS3) protein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Affinities of the selected phage clones to anti-NS3 monoclonal antibody 3B1C4 were measured by phage ELISA. The results are expressed as the mean optical density (OD492) obtained with triplicate of phage clones tested, at the final dilution of 1.0e11 phages/ml with 3B1C4 coated on the solid phase minus the mean OD of triplicate of the same phage dilutions tested with an irrelevant mAb. 2935 KPPNPSPMSRFI(6/49) VKIPNYPPNSTS(5/49) VTIPQYSPRHPV(3/49) QKPPNPSPISPL(2/49) VKIPQHINLTSE(2/49) ISPYNTIVAKLR(2/49) KPPNPSPIEHWP(1/49) KPPNPSPLALAG(1/49) KPPNPSHLSLTW(1/49) KPPNPHPLPYES(1/49) KPPNPPPPPHDL(1/49) LKPPNPMPPAHG(1/49) VKIPNTYRLGMA(1/49) VKIPNSGTALSR(1/49) VKIQNNPPPLPQ(1/49) VKINNSSPLPTG(1/49) VKIQNLPTLNTK(1/49) VKIPQFLASPLA(1/49) VKISQYASMPPI(1/49) VKIPQHMHPLPI(1/49) AKIDNAVPQTVS(1/49) LKIPNSPSRTVG(1/49) LKIPSKFSPAML(1/49) VRIPNPPPTPFL(1/49) VRIANHPPEPFR(1/49) IRIPQHDWPSGM(1/49) VTIPQYSTPPSV(1/49) VTIANNPIFTSV(1/49) ITIPQKQEYAYT(1/49) VSISNLGPQYNT(1/49) VSIPNTWPSTKT(1/49) VMISNDSKGRWW(1/49) DINLVYNTVLNK(1/49) KPPNPRPTMSLW(1/49) TKPPNPKPSMFF(1/49) 35 Phage display (subtractive panning) 3 PMID:15272409 Serum immunoglobulin (Ig) G antibodies from patients with SARS Human SARS coronavirus Not determined. Not determined. Ph.D.-12 phage display library (X12) To enhance selection of peptides binding to IgG specifically associated with patients with SARS, we designed a preclearing step to remove nonspecific clones by preabsorption of the phage-displayed 12-mer peptide library onto purified IgG from normal serum pooled from 5 control subjects. 2936 LTANTKSMALSG(20/35)[0.44] AQLPIPRWNFMS(4/35)[0.40] STLAKHAPAGYR(2/35)[0.43] LTTDHSTTVING(2/35)[0.47] VPPNTKSMALSG(1/35)[0.47] LTTHTKSMALSG(1/35)[0.40] VPANIKSMALSG(1/35)[0.37] VAPSMAkSIPAK(1/35)[0.51] AHLHIPRWNLMS(1/35)[0.24] KTNNSYIPPLTW(1/35)[0.53] ISQSLMPLSPWR(1/35)[0.51] T-K-S-M-A-L-S-G, I-P-R-W-N-[FL]-M-S 11 Phage display (common panning) 4 PMID:15607634 Anti-burkholderia pseudomallei protease polyclonal antibody IgG Burkholderia pseudomallei protease Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Affinities of the positive phage clones to burkholderia pseudomallei protease polyclonal antibody IgG were measured by phage ELISA. The A405 values were measured and data shown were reproduced from the Fig. 3a in the reference. wild-type M13 phage was used as control, which had the value of 0.27 when that phage binded to the target. 2937 ATTVPAS FNQNPRS SVLTLET TLMLRET RPPTRYH KAWIIPA 6 Phage display (competitive panning) 3 PMID:16037945 Anti-LKM1 autoantibodies from hepatitis C–infected patient 1 Cytochrome P450 2D6 (CYP2D6) protein Not determined. Not determined. Ph.D.-7 phage display library (X7) The retained phages, eluted with CYP2D6 fusion protein and purified, were then sequenced. 2938 RALSMPP STVMTRP YSPLTLI FPLHLGV TSQPTHT SSTHSAP STQHGPQ 7 Phage display (competitive panning) 3 PMID:16037945 Anti-LKM1 autoantibodies from hepatitis C–infected patient 2 Cytochrome P450 2D6 (CYP2D6) protein Not determined. Not determined. Ph.D.-7 phage display library (X7) The retained phages, eluted with CYP2D6 fusion protein and purified, were then sequenced. 2939 ESLPTFA AIDTRPW SYHSDNN NYNWTTP QVMPYVQ GPHPRSV 6 Phage display (competitive panning) 3 PMID:16037945 Anti-LKM1 autoantibodies from hepatitis C–infected patient 3 Cytochrome P450 2D6 (CYP2D6) protein Not determined. Not determined. Ph.D.-7 phage display library (X7) The retained phages, eluted with CYP2D6 fusion protein and purified, were then sequenced. 2940 QDQRTFT LVTHTPI VPATLFR PHLHPPR MSLHHSH ALRTASS HAIYPRH RPPPTAT 8 Phage display (common panning) 3 PMID:26640539 Duck hepatitis B virus polymerase (DHBVP) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2941 CPTSHHRAC(8) CKQPSHSAC(2) CTANSSAQC(2) 3 Phage display (in vivo) 4 PMID:26568284 Sheep intestinal lymph Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Phages preselected by 2 rounds of in vitro panning were amplified and further subjected to 2 rounds of in vivo panning procedure in sheep. 2942 CTANSSAQC(12) 1 Phage display (in vivo) 4 PMID:26568284 Cells from sheep lymph plasma Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Phages preselected by 2 rounds of in vitro panning were amplified and further subjected to 2 rounds of in vivo panning procedure in sheep. 2943 STVMDSAVIVKS(8) 1 Phage display (in vivo) 4 PMID:26568284 Epithelial layer overlaying Peyer’s Patches (PP) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phages preselected by 2 rounds of in vitro panning were amplified and further subjected to 2 rounds of in vivo panning procedure in sheep. 2944 CDPSPSRHC(1) 1 Phage display (in vivo) 4 PMID:26568284 Epithelial layer overlaying Peyer’s Patches (PP) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Phages preselected by 2 rounds of in vitro panning were amplified and further subjected to 2 rounds of in vivo panning procedure in sheep. 2945 SNLSWPANMKHP(23/63) TWHHSYFNSVSV(22/63) ATTPNDNDLNRW(3/63) FHESWPSMSSAA(3/63) RDPYTLIQYLSV(2/63) MVFPKDGREAKL(2/63) WNAPPMINRMST(1/63) TWFEALRLNTDM(1/63) VETPTLRYSHWQ(1/63) TGHHSYFNSVSV(1/63) EYSARVQYLQFR(1/63) HAGQKDLLSAWM(1/63) VDLDFRSLILVQ(1/63) NMTPFFEGVIFN(1/63) 14 Phage display (common panning) 3 PMID:26624919 Anti-pkMSP-142 antibody Merozoitesurface protein-142(MSP-142) Not determined. Not determined. Ph.D.-12 phage display library (X12) 2946 APDTKTQ(14)[0.96 ± 0.02] ATRQPNH(6)[0.83 ± 0.10] HASKQLL(5)[0.71 ± 0.13] SPRVGAT(4)[0.72 ± 0.14] LSLSTTS(4)[0.70 ± 0.16] HLHAHKL(4)[0.74 ± 0.11] RHFHFPA(2)[0.79 ± 0.10] HPMSAPR(1)[0.81 ± 0.09] 8 Phage display (common panning) 4 PMID:26496918 Human soluble receptor for advanced glycation end products (sRAGE) Amyloid β (Aβ) peptide Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding activities of phage clones to the sRAGE were measured by phage ELISA. The OD values at 370 nm were measured and data shown were reproduced from the Fig. 1 in the reference. 2947 WTFNAGSHERTY(7/15)[1.24 ± 0.07] WTLQQEEQLVYY(1/15)[1.38 ± 0.12] WQLQSGEELRLT(1/15)[1.39 ± 0.11] AVEWGAGMPSYS(1/15)[1.04 ± 0.12] WGTYQWGTEAPN(1/15)[1.11 ± 0.10] WSLQAGENVTIF(1/15)[1.09 ± 0.18] NVPAPVALYQIS(1/15)[0.18 ± 0.04] PLYGDDANHYQW(1/15)[0.50 ± 0.36] SNWPTSRTYLSD(1/15)[0.29 ± 0.07] 9 Phage display (common panning) 3 PMID:26541335 Anti-plant potyviruses capsid proteins (CPs) monoclonal antibody C4 Capsid proteins (CPs) of plant potyviruses Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of the positive phage clones to the C4 MAb were measured by phage ELISA. The OD values at 405 nm were measured and data shown were reproduced from the Fig. 1 in the reference. 2948 LPLTPLP(6)[NT][NT] FHGAREM(4)[0.45 ± 0.01][0.36 ± 0.02] TTYSRFP(1)[NT][NT] NEISFHA(1)[0.62 ± 0.02][0.38 ± 0.03] LPLGHHE(1)[NT][NT] IGHLSFE(1)[NT][NT] GHLYDDP(1)[0.74 ± 0.02][0.64 ± 0.01] 7 Phage display (subtractive panning) 3 PMID:26543535 Maltase-glucoamylase (MGAM) Anti-MGAM polyclonal antibody Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding activities of the positive phage clones to the maltase-glucoamylase were measured by phage ELISA and competitive phage ELISA. The A450 values were measured and data shown were reproduced from the Fig. 1a in the reference. In the competitive phage ELISA, a competitive inhibitor (0.5 mmol/L acarbose and 0.5 mmol/L voglibose) was added. M13 phage vector showing no peptide was used as control, and the corresponding affinity values were 0.17 ± 0.01 and 0.08 ± 0.01 respectively. NT means not test. In the second and third round of selection, an aliquot of amplified eluate from a previous round of biopanning was added to anti-MGAM antibody-coated beads. After 30 min of gentle shaking at room temperature, the supernatant was transferred to the enzyme-coated beads and was incubated for 60 min. 2949 CTHYGFRGC(2)[1.12 ± 0.01][0.79 ± 0.02] CGHHHRDYC(1)[1.07 ± 0.07][0.59 ± 0.02] 2 Phage display (subtractive panning) 3 PMID:26543535 Maltase-glucoamylase (MGAM) Anti-MGAM polyclonal antibody Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The binding activities of the positive phage clones to the maltase-glucoamylase were measured by phage ELISA and competitive phage ELISA. The A450 values were measured and data shown were reproduced from the Fig. 1a in the reference. In the competitive phage ELISA, a competitive inhibitor (0.5 mmol/L acarbose and 0.5 mmol/L voglibose) was added. M13 phage vector showing no peptide was used as control, and the corresponding affinity values were 0.17 ± 0.01 and 0.08 ± 0.01 respectively. NT means not test. In the second and third round of selection, an aliquot of amplified eluate from a previous round of biopanning was added to anti-MGAM antibody-coated beads. After 30 min of gentle shaking at room temperature, the supernatant was transferred to the enzyme-coated beads and was incubated for 60 min. 2950 RDGSIAMHSMIP(1)[0.36 ± 0.01][0.27 ± 0.02] QALLEGNAKGGN(1)[NT][NT] GGTKTHVDFSLK(1)[NT][NT] 3 Phage display (subtractive panning) 3 PMID:26543535 Maltase-glucoamylase (MGAM) Anti-MGAM polyclonal antibody Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of the positive phage clones to the maltase-glucoamylase were measured by phage ELISA and competitive phage ELISA. The A450 values were measured and data shown were reproduced from the Fig. 1a in the reference. In the competitive phage ELISA, a competitive inhibitor (0.5 mmol/L acarbose and 0.5 mmol/L voglibose) was added. M13 phage vector showing no peptide was used as control, and the corresponding affinity values were 0.17 ± 0.01 and 0.08 ± 0.01 respectively. NT means not test. In the second and third round of selection, an aliquot of amplified eluate from a previous round of biopanning was added to anti-MGAM antibody-coated beads. After 30 min of gentle shaking at room temperature, the supernatant was transferred to the enzyme-coated beads and was incubated for 60 min. 2951 AELAKLPLF GAISFDHVHP 2 Phage display (subtractive panning) 6 PMID:26543513 Colorectal cancer cell(HCT-8) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection using magnetic beads to capture M13 phage display peptides that were not of interest was first performed such that peptides binding on non-target cells (control cells) could be removed. Then, the supernatant was collected to perform the subsequent positive selection. As mentioned previously, the target cells surfaced-coated with magnetic beads were incubated with the supernatant to perform the positive selection. 2952 MNAKWPIENLMN 1 Phage display (subtractive panning) 6 PMID:26543513 Cancer stem cell(CSCs) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Negative selection using magnetic beads to capture M13 phage display peptides that were not of interest was first performed such that peptides binding on non-target cells (control cells) could be removed. Then, the supernatant was collected to perform the subsequent positive selection. As mentioned previously, the target cells surfaced-coated with magnetic beads were incubated with the supernatant to perform the positive selection. 2953 SVACHSTKAHSC RVACLYHTTRAC VCACLYHTTRAC VCACNWNNHRAC FTACNWNNHRAC RRACQSHKARSC 6 Phage display (common panning) 3 PMID:26438313 Human squalene synthase(hSQS) BPH701 (1-phospho-4- (3-(4-pyrolphenoxy) phenyl) butane-1-sulfonic acid Not determined. Not determined. Ph.D.-12 phage display library (X12) 2954 DHIHRSYRGEFD(3/16) NIYTTPWGSNWS(2/16) SHSLPASADLRR(2/16) SAAYLAVIDTSS(1/16) HSPQYWVHHWRG(1/16) GQSEHHMRVASF(1/16) YQFGYDYPRSQV(1/16) WSGPAVMLETFF(1/16) ERLSDFANFRAP(1/16) HYPFMTPVLFTP(1/16) SALKGLFPADHH(1/16) AAFPSLNLRTQP(1/16) 12 Phage display (common panning) 3 PMID:26521981 Beta-lactamase 2 (EC:3.5.2.6) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2955 ALWPPNLHAWVP(9) AHSANNFNVKGI(3) FTTKTYNGVAER(2) ALWPPNPQALVP(1) NHSVNNLNVKGI(1) VALKAWAHPTHW(1) FPRNVANSPVYN(1) YSAHNYIGNSGP(1) HVVKQAMSNNMM(1) RFWPPNLQAGVP(1) RLNASSMISGRV(1) 11 Phage display (common panning) 3 PMID:26482679 Anti-DNA polyclonal antibody IgG1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2956 ALWPPNLHAWVP(5) TPMVERNYNAAN(3) RLNASSMISGRV(3) AHSANNFNVKGI(2) ALWPPIPHAWVH(1) KLLLPIPQVGPL(1) KLLSPNLQAGVL(1) KLLFAIPEVEGF(1) RLNNSSMNSGRV(1) NWNNPPIRLVAS(1) YSAHNYIGNSGP(1) NSVHTNTGNLGL(1) TPPNNYYYVEAV(1) IGSPHLTAPLLT(1) WHSWAMHQNEHF(1) HNGGAMHIQKMK(1) HVLYTIAGGLEP(1) LNWTPASTSLKN(1) NQAQFELLNKVK(1) 19 Phage display (common panning) 3 PMID:26482679 Anti-DNA polyclonal antibody IgG2a Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2957 ALWPPNLHAWVP(11) ALWPPNLQAWAP(1) ALWPPNLQAGVP(1) KLWLPNLQAGVP(1) AFYPNNTVSRLA(1) AHSANNFNVKGI(1) NHLANNFNNKGI(1) THNFRFLMTLGW(1) TNQNYTSLPVAA(1) HVVKQAMSNNML(1) HVVKQAMFHNMM(1) VGANWSSYHVLN(1) NNLNYTSNPVAH(1) NSNWKTFGGWPS(1) YNHSATHLVSLP(1) TPTSTLHKFKHW(1) SSPPISMGSVGG(1) RIILRVLLRCIR(1) 18 Phage display (common panning) 3 PMID:26482679 Anti-DNA polyclonal antibody IgG2b Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2958 ALWPPNLHAWVP(16) AHSANNFNVKGI(6) HVVKQAMSNNMM(4) KNMAKAENNVYM(3) SCTGLLSYHSRC(3) VHLQAGELMNPK(2) AHSAANFNVKGW(1) SYSALNHPPHPT(1) NLSHFLPLSAPV(1) NYEFSISEQSHN(1) AETVESCLAKPH(1) APTTQRVFWNQS(1) 12 Phage display (common panning) 3 PMID:26482679 Anti-DNA polyclonal antibody IgG3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2959 SWQIGGN(6)[1.10 ± 0.11] IGNSNTL(2)[1.08 ± 0.09] QFHFDAP(1)[1.22 ± 0.12] TSPFVVP(1)[1.13 ± 0.07] TGNSNTQ(1)[1.24 ± 0.13] TSHFEVP(1)[1.20 ± 0.05] 6 Phage display (subtractive panning) 4 PMID:26555399 Human ovarian cancer cell line HO8910 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The binding activities of the positive phage clones to human ovarian cancer cell line HO8910 were measured by phage ELISA. The OD values at 450 nm were measured and data shown were reproduced from the Fig. 1 in the reference. M13KE was used as controls and had the affinity value 0.19 ± 0.02 with this target. Biopanning began with the incubation of the phage display peptide library with both normal and tumor cells, in which normal cells were used to deplete peptides that only bind to normal cells and then further incubated with tumor cells for identifying the peptides that only specifically bind to tumor cells. Peptide1 read as SWQIGGN was the positive peptide and showed preferential binding to the target cells. Peptide 1 also inhibited cell proliferation, migration, invasion and adhesion of ovarian cancer HO8910 cells in vitro. 2960 THENWPA(16/20)[1.15 ± 0.12] APQWPAN(3/20)[0.77 ± 0.11] DVTSNFP(1/20)[0.74 ± 0.06] 3 Phage display (subtractive panning) 4 PMID:26423339 CD44v3-v10 glycoprotein Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding activities of the positive phage clones to CD44v3-v10 glycoprotein were measured by phage ELISA. The OD values at 450 nm were measured and data shown were reproduced from the Fig. 2a in the reference. Data are presented as the mean ± SD. Unrelated phages randomly selected from the original library served as the negative control and had the affinity value 0.27 ± 0.03 with this target. The library was incubated with the CD44s to eliminate phages binding to CD44s. Next, unbound phages were incubated with CD44v3-v10 and bound phages were eluted with 0.2 M glycine-HCl (pH 2.2) supplemented with 1% (w/v) BSA. 2961 NTWHNSR(12/34)[0.33 ± 0.03] WHKEQFW(9/34)[0.12 ± 0.02] QTKMTNP(2/34)[0.20 ± 0.02] TVKTRPA(2/34)[0.29 ± 0.02] AGPTRIS(2/34)[0.11 ± 0.05] THSPGLL(1/34)[0.23 ± 0.02] SINPDNR(1/34)[0.20 ± 0.02] ALLADSR(1/34)[0.22 ± 0.04] EIALGAR(1/34)[0.22 ± 0.03] ATSAIHG(1/34)[0.14 ± 0.02] HVQLLIF(1/34)[1.89 ± 0.03] SITTVAA(1/34)[0.13 ± 0.02] 12 Phage display (competitive panning) 4 PMID:26577672 Immunoglobulin G (IgG) of Tuberculosis (TB) patients Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding activities of the positive phage clones to IgG from TB patients were measured by phage ELISA. The A450 values were measured and data shown were reproduced from the Fig. 1a in the reference. Data are presented as the mean ± SD. The first eluted phage library was used as negative control and had the affinity value 0.39 ± 0.03 with this target. Bound phage was eluted with free purified IgG from TB patients to compete the bound phage away from the immobilized antibodies on the plate. 2962 TTAGIRQ(19/34) SNAPQKL(8/34) FHFPPVY(2/34) WHLPLSL(1/34) ANVWMPT(1/34) LPPPPPFS(1/34) WHALPTN(1/34) AHGPLAF(1/34) 8 Phage display (subtractive panning) 4 PMID:26577672 Immunoglobulin G (IgG) of Tuberculosis (TB) patients Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding activities of the positive phage clones to IgG from TB patients were measured by phage ELISA. The A450 values were measured and data shown were reproduced from the Fig. 1b in the reference. Data are presented as the mean ± SD. The first eluted phage library was used as negative control and had the affinity value 0.06 ± 0.01 with this target. The library was added onto the healthy IgG coated plate first, and the unbound phage supernatant was added to TB patients IgG package plate. 2963 EHFSTGMG DGLRPQAP DGLRPPGS DGLRSPPG DGLRSPGS DGLRNPSS D-G-L-R 6 Phage display (common panning) 3 PMID:26456116 Anti-Gonadotropin releasing hormone (GnRH) polyclonal antibody of cat 1 Gonadotropin releasing hormone (GnRH) Not determined. Not determined. f8-8mer phage display library (X8) 2964 EHPSYGLA DHPSYTPM ETSYADHT H-x-S-Y 3 Phage display (common panning) 3 PMID:26456116 Anti-Gonadotropin releasing hormone (GnRH) polyclonal antibody of cat 2 Gonadotropin releasing hormone (GnRH) Not determined. Not determined. f8-8mer phage display library (X8) 2965 DAPAHWSQ EPTSHWSA EPSPHWSS ETTSHWSS DDTHWAAP EDYGARPA DSDYGARP DTDYGARP EDSYGERP ESASYGER ESVSYGAK ESASYGER ESDYGARV EEYGARST EEYGMRGP EAGYGEK H-W-S, Y-G-x-[RK]-P 16 Phage display (common panning) 3 PMID:26456116 Anti-Gonadotropin releasing hormone (GnRH) polyclonal antibody of dog Gonadotropin releasing hormone (GnRH) Not determined. Not determined. f8-8mer phage display library (X8) 2966 EGLRPSGA EGLRPSNA DGLRPSNS EGLRPSGQ EGLRPSGS EGLRPTGS EGLRPSGQ [ED]-G-L-R-P-S 7 Phage display (common panning) 3 PMID:26456116 Anti-Gonadotropin releasing hormone (GnRH) polyclonal antibody of rabbit Gonadotropin releasing hormone (GnRH) Not determined. Not determined. f8-8mer phage display library (X8) 2967 SYLPETIYEYRL(26) VENKTRYHDREV(7) HEGAWHNYARSV(1) 3 Phage display (common panning) 3 PMID:26709700 FvTox1 protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2968 SNGRVAD(1) 1 Phage display (common panning) 3 PMID:26709700 FvTox1 protein Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2969 RMLPASLHL RRLPEELHR ARLPPSLHL RNLPRALSP RRLPSAFHP LRSLHLVRG SRPPSLFRP RNVFLIRTE GRVFFLVRP DRNIYLVRQ 10 Phage display (common panning) 2 PMID:12943796 Anti-HBX monoclonal antibody Recombinant HBX protein Not determined. Not determined. pVIII-9aa phage display library (X9) 2970 LSLTSTL LPTPALR ASGLLSL SLTSTLY TLLVQGS LPLNSSA LSLPLNS SSSLTLK VTSALSR SALSRDG RLSVPPL GLLSLTS GSPSTPA LSLSSIT SLTLKVT KVLSLPL SLPLNSS LSVPPLV 18 Phage display (common panning) PMID:27143691 Human KRAS-mutant colorectal cancer cell lines and tissues Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The ex-vivo screenings were run on pooled specimens from different animals (n=3), with a standard procedure that involved two successive selection rounds (R1, R2) and two alternative controls (C1, pseudo-metastasis from WT cells; C2, non-tumoral liver). The in vitro screenings were performed with the same standard procedure (P1) or with an enrichment protocol (P2) in which several rounds were iterated until increased phage binding to KRAS mutant vs WT cells was observed. Each KRAS mutational status (G12D, G12V, G13V) was independently screened in 4 biological replicates, i.e. 2 cell lines (SW-48, LIM1215) × 2 clones, and in 2 experimental conditions (ex-vivo and in vitro) for a total of 24 phage display screening experiments. Each of them consisted of 5 output samples (3 mutant KRAS + 2 controls), giving a total of 120 samples. This subset of peptides has been derived from a larger, deep-seq analysis. These peptides are identical to regions of the human BCAM protein and are potential ligands for LAMA5. 2971 RRPRMLT[1.40 ± 0.23] QLRRQRR[NT] HSRRMRK[NT] RSRRIRL[5.37 ± 0.49] R-R-x-R 9 Phage display (common panning) 4 PMID:17699853 Neuropilin-1 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Surface plasmon resonance (SPR) Surface plasmon resonance was used to measure the real-time association and dissociation of the binding of RRXR-containing peptides to NRP1. The average dissociation constants (KD) for the binding of DG1 and DG2 to NRP1 were 1.40 ± 0.23 and 5.37 ± 0.49 umol/L, respectively. Two potent synthetic anti-NRP1 peptides, CRRPRMLTC and CRSRRIRLC, which block NRP1 signaling pathways and suppress tumorigenesis, cancer invasion, and angiogenesis, were identified. 2972 SPFWWSD(25) SPFWWTD(36) PNPFVLD(1) LGSVQHT(1) SPFWWSL(1) FSPFFWN(1) WILFFLD(1) NSPFQLF(1) PGLATNF(1) 9 Phage display (subtractive panning) 3 PMID:26806845 Purified IgG from serum of juvenile idiopathic arthritis (JIA) persistent oligoarticular subtype Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) For screening the PD library to select JIA mimotopes, a subtractive step against purified IgG from healthy children and patients with other autoimmune diseases, followed bya positive selection against purified IgG from JIA subtypes was used. 2973 SPFWWHQ(3) SPFFLTP(1) WNPFLLD(1) QSPFHLF(1) NPGWLGS(1) SAVIKSS(1) SSHLLSE(1) LPSRSQT(1) NNPFQLW(1) SSFWWTD(1) 10 Phage display (subtractive panning) 3 PMID:26806845 Purified IgG from serum of juvenile idiopathic arthritis (JIA) extended oligoarticular subtype Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) For screening the PD library to select JIA mimotopes, a subtractive step against purified IgG from healthy children and patients with other autoimmune diseases, followed bya positive selection against purified IgG from JIA subtypes was used. 2974 FDPFGWS(2) SSWLPRG(1) WSPFLAP(1) SPFDWWF(1) NPFFLTA(1) SPNPPLH(1) SSPFFLW(1) SPFHLLP(1) 8 Phage display (subtractive panning) 3 PMID:26806845 Purified IgG from serum of juvenile idiopathic arthritis (JIA) polyarticular rheumatoid factor negative subtype Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) For screening the PD library to select JIA mimotopes, a subtractive step against purified IgG from healthy children and patients with other autoimmune diseases, followed bya positive selection against purified IgG from JIA subtypes was used. 2975 NPFSLLA(1) YFTAPPD(1) DSPFRLW(1) TRPTASH(1) SESPGIA(1) FSPFFAP(1) SPFWLAA(1) SPFFLGP(1) TPFWFLD(1) PAPSRSQ(1) RFGGLIA(1) 11 Phage display (subtractive panning) 3 PMID:26806845 Purified IgG from serum of juvenile idiopathic arthritis (JIA) polyarticular rheumatoid factor positive subtype Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) For screening the PD library to select JIA mimotopes, a subtractive step against purified IgG from healthy children and patients with other autoimmune diseases, followed bya positive selection against purified IgG from JIA subtypes was used. 2976 CQMHQLSSC CSGMKTTGC CLSPHSMFC CPWWYGPWC CLGVHSSSC CSTPWHQWC CHQHNSMYC CKTENMQSC CSTLKVATC CTVNWYPLC 10 Phage display (common panning) 3 PMID:26922723 Human squalene synthase(hSQS) BPH701 (1-phospho-4- (3-(4-pyrolphenoxy) phenyl) butane-1-sulfonic acid Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2977 VVPWAGL WTVHTLS HMFPWRQ QSWLPSL GYKDFSA RILITIP WASKVAR 7 Phage display (common panning) 3 PMID:26896129 Hemagglutinin-esterase (HE) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2978 TPARHIY TLLRVDN GSGNAFM MPLGFKA NERALTL VQPIPAT 6 Phage display (common panning) 3 PMID:26896129 Fusion glycoprotein F0 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2979 CGSPAPNAC CALIPPLLC CRVTLPPHC CLPSTFRGC CLPALARSC CSPLSVLSC CVHSPMRLC CTPSGHSAC CSPSFGPRC CSWSALFGC CSTVSPFLC CPSIYPLLC CGIPERSSC 13 Phage display (subtractive panning) 3 PMID:26762591 Mucin-5AC Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) After its pre-incubation within a well blocked with the blocking buffer followed by a pre-incubation within a well coated with 100 μL of MUC2, the peptide library was incubated within a well coated with MUC5AC. 2980 LVRPLAL(15/16) KPHQGLI(1/16) 2 Phage display (competitive panning) 4 PMID:26833245 Hemagglutinin, HA Ganglioside GM3 Not determined. Not determined. Ph.D.-7 phage display library (X7) GM3, one of the receptors of HA, was used to obtain phages that bound with the receptor-binding site of HA. 2981 STQHHHHSKQSR(32) HWKPHSNLHLSR(8) WPGHHNHSMKHK(6) HLKHTHNTHYKT(4) 4 Phage display (subtractive panning) 3 PMID:26913962 Glycoprotein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Biotin was added to displace any phages bound to the streptavidin and the plates were washed 15 times with TBST to remove non-binding phages. 2982 HFPSPIFQRHSH(6/34) YHPNGMNPYTKA(4/34) QHRFANHLIFKT(1/34) WHKPWMFGKLTQ(1/34) HRQNVSQPVNPQ(1/34) HPSSPTPSPWRF(1/34) LSATSRLQFPSI(1/34) HYKYYPTASVMK(1/34) HFNRDWQKIHGP(1/34) HPIWYPTNINRQ(1/34) KVWTIDTAHTRA(1/34) GAFHVWQPTVTM(1/34) HQSKISIAVDQP(1/34) SSWWGEGMNKSY(1/34) 14 Phage display (common panning) 4 PMID:26903196 Milled wood lignins Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2983 KIWIPPKPMSPW(1/34) LAPRHTHSIHPS(1/34) MHTQRTPWIFSL(1/34) SMGPTRTPPPNT(1/34) HSAKLWLIPSMS(1/34) GLKVWTVQPPHV(1/34) YTQVPTKMQLGG(1/34) QMKCCIATYNPP(1/34) SYGLKPFMPWYS(1/34) VNNHWDNSHPNT(1/34) QMSITLQNSYLI(1/34) SLDAWEVERRST(1/34) 12 Phage display (common panning) 3 PMID:26903196 Milled wood lignins Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2984 VQHNTKYSVVIR(4/36) KGPHFPSPHVAL(2/36) YHPNGMNPYTKA(2/36) SHEPVLMIQKFK(2/36) VQHNRKYSVVIR(1/36) WTPNKLKTMQVK(1/36) AITHGAKMPAKI(1/36) LPTKTLYPHVRM(1/36) STVKYHNHNRNF(1/36) VPHMAPHRIAAQ(1/36) FATNHRTTHERI(1/36) DHAARNWVERQR(1/36) VQHNTKYSDVIR(1/36) VQHHHSYKGVTY(1/36) 14 Phage display (common panning) 4 PMID:26903196 Milled wood lignins Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2985 SKMAHMERSWEV(2/36) MNRHSSLPLKPW(1/36) AHNVMVATKIPK(1/36) LTLNKHPNSHHI(1/36) KLSNFHPQGSMM(1/36) THLGLYQRNTMN(1/36) SKHNYPSQGPVF(1/36) QKTNHHAHIWDG(1/36) HHGWVSPQYGVA(1/36) LPKHNEHYFTMP(1/36) LVNYQSELHQTR(1/36) DMKWTLKEWMTH(1/36) GQHNTNGNQTIT(1/36) VSLNTDYWNRNY(1/36) QEQDWTNSQRIN(1/36) 15 Phage display (common panning) 3 PMID:26903196 Milled wood lignins Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 2986 HYIDFRW(2/24) GHDPTPL(2/24) TVNFKLY(1/24) 3 Phage display (common panning) 3 PMID:26896661 Fe3O4 nanoparticles Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2987 TVNFKLY(6/37) HYIDFRW(1/37) TSTGAIA(1/37) APFNFGN(1/37) VPASPWT(1/37) GQSEKHL(1/37) MNSNIPI(1/37) 7 Phage display (common panning) 4 PMID:26896661 Fe3O4 nanoparticles Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2988 LNLQTQL(12/50) HYIDFRW(7/50) TVNFKLY(4/50) GQSEKHL(3/50) MNSNIPI(3/50) QLAVAPS(3/50) TSTGAIA(2/50) GVEGHKP(2/50) APFNFGN(1/50) VPASPWT(1/50) 10 Phage display (common panning) 5 PMID:26896661 Fe3O4 nanoparticles Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 2989 TSRTPLHKP RAGGFEKHS 2 Phage display (common panning) 2 PMID:26739405 Anti-birch pollen HMW allergens monoclonal antibody BIP3 Pollen allergens Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) 2990 CHKLRCDKAIA(3) WRPRWLYD(1) 2 Phage display (common panning) 3 PMID:26739405 Anti-birch pollen HMW allergens monoclonal antibody BIP3 Pollen allergens Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) 2991 CHKLRCDKAIA(6) CKASSCDTGHC(1) CFFAWRSLPNCP(1) 3 Phage display (common panning) 4 PMID:26739405 Anti-birch pollen HMW allergens monoclonal antibody BIP3 Pollen allergens Not determined. Not determined. pVIII-9aa.Cys phage display library (CX9C) 2992 HVDMIPR(29) ALPRIHN(27) FPLMGHG(23) GFATITG(21) EVLPYRE(20) EVPILRS(17) HIYTALV(12) AIYPNTY(4) 8 Phage display (subtractive panning) 5 PMID:26711806 Nogo-66 receptor (NgR1) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The plates incubated with NgR1 were washed and followed by blocking with Nogo-66 for 12 h at room temperature. After washing with 0.05% TBST, the phages from the fourth round were added to the plates and the unbound positive phage were collected. 2993 DHLASLWWGTEL(70%) DWSSWVYRDPQT(22.5%) EHFSLWWNPPLV(5%) NHFSYEFWFSLP(2.5%) 4 Phage display (competitive panning) 5 PMID:26850086 Glypican-3 (GPC3) protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The fifth round of panning procedure was slightly altered by using human recombinant GPC3 protein (100 μg/mL) for competing off the GPC3 binding peptide. 2994 GMVQTIF(8/16) CMTCLLA(2/16) GMVQTIW(2/16) AAISGWW(1/16) CMTCLLR(1/16) CMTCLIR(2/16) 6 Phage display (competitive panning) 4 PMID:26695281 Anti-OchratoxinA(OTA) monoclonal antibody 2A11 Ochratoxin A (OTA) Not determined. Not determined. Ph.D.-7 phage display library (X7) In the fourth round of panning, the elution buffer was a PBS solution containing 10% (v/v) methanol and 0.1 ng/mL OTA. 2995 SWLAMEF GGNPTIY MNPTYTM NVAPAMS QVRSVVH DQTQTPR DSKHNFS MGPPRTS TPIANPP YTSTGPA GTLWSSM SHGRINS HVLVPLH NMIPSHG TDTASRS 15 Phage display (subtractive panning) 4 PMID:27038522 α1β glycine receptor Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Subtractive panning consisted of washing the phage library over negative selection HEK293 cells expressing α2β glycine receptors. 2996 STIHGST SIRLDSQ HAPKSDT DRMPHYF STFTKSP TGADLNT YTMPGEL VIPHVLS GVQIMGR NANAALP SWQQGPY GSSCCKT SILPYPY KLPGWSG NQLTTLN AAPTVPR QETRAPG NQLPLHA GPMLARG TTMPIDS TTPTKSA VQTYARG TSLNRYP SHTAPLR DVPVPQV 25 Phage display (subtractive panning) 4 PMID:27038522 α1β glycine receptor Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Subtractive panning consisted of a pair of washes of library over negative selection HEK293 cells expressing α2β, followed by HEK 293 cells expressing α3β glycine receptors. 2997 MAAKYN(85.7%) MYVIRG(7.1%) SLSWVC(2.3%) QRKMAS(2.3%) MSKPKQ(2.3%) MAHYSG(2.3%) 6 Phage display (common panning) 4 PMID:26995695 Leishmania major metacyclic parasite Not determined. Not determined. Not determined. X6 fdMED1 phage display library 2998 CKIPILANC CLSPTHMLC CTVHPRSLC CSTSPEHAC CSAPWNHAC CSAHNWPNC CSARFTYTC CSTYELSEC CMPWQKNRC CMPVWKPDC CVWYLSPYC CPDQYNVHC 12 Phage display (competitive panning) 3 PMID:26976271 Heterotrimeric tRNA-dependent amidotransferase (GatCAB) Glu-tRNAGln Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) In the third round, Glu-tRNAGln was used to specifically elute phages bound to GatCAB. 2999 NFMESLPRLGMH(8)[2.682 ± 0.052, 25.8] WHWTWPSEYPPP(4)[2.734 ± 0.064, 20.6] YPWPWWHSVSPW(3)[2.014 ± 0.159, NT] WHWTRLSEYPPP(3)[1.918 ± 0.042, NT] WHWTWLSEYPPP(2)[2.406 ± 0.138, NT] WHWTGLSEYPPP(2)[1.124 ± 0.063, NT] FHWHPWFTSVQY(2)[2.343 ± 0.137, NT] SLKIRSPLQLPK(1)[NT, NT] SVSVGMKPSPRP(1)[NT, NT] HFSSWMWAPSWT(1)[NT, NT] 10 Phage display (common panning) 4 PMID:27220907 Lipopolysaccharide, LPS Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA,Surface plasmon resonance (SPR) Affinities of isolated phage clones to S. Typhimurium were measured by phage ELISA. The plates were read at 450 nm using microplate reader (Synergy, Bio-Tek, USA). The experiments were performed in triplicates. Data shown (the first column) were reproduced from the graph. To further validate the ELISA results, Surface Plasmon Resonance analysis was performed for WHWTWPSEYPPP (pep38) and NFMESLPRLGMH (pep49) using Biacore T-200 (GE Healthcare Ltd., India). The SPR analysis was used to quantify the affinity of peptide towards its target, by calculating the dissociation constant (Kd, nM). The dissociation constants KD (μM) were shown (the second column). NT denotes not tested. The peptide NFMESLPRLGMH (pep49) derived from biopanning displayed a high affinity (25.8 nM) for the LPS of S. Typhimurium and low cross-reactivity with other strains of Salmonella and related Gram-negative bacteria. Further, pep49 was able to detect S. Typhimurium at a LOD of 1.0e3 CFU/mL using ELISA, and may be a potential cost efficient alternative to antibodies. 3000 CKMTRSTIC(29) CKIMISISC(19) CLIRRTSIC(8) CPLLTKLKC(8) CHIPSIHSC(8) CTHRRSTPC(7) CLHPPLTLC(6) CNLLLRTQC(5) CQMMLIRRC(4) CPRRSHPIC(4) CILRKIRPC(4) CKMNIRLSC(2) CSRRPTSIC(1) CLIRLILQC(1) CRMRKTLRC(1) CQPHHSIIC(1) CSTHIPSHC(1) CILRRRKRC(1) CSIRIHRRC(1) CSLRRHPIC(1) CRTKLRKLC(1) CHMQIMHRC(1) CTRMMLRIC(1) CTPRQTRMC(1) CMLNITRRC(1) CRKLPRISC(1) CPLQPKSIC(1) CPIRHRPIC(1) CRTMRIRLC(1) 29 Phage display (in vivo) 3 PMID:27173134 Endothelial cells (ECs) of ligated left carotid artery (LCA) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Briefly, 1.0e11 plaque-forming units (pfu) of phages were injected intravenously into mice at 3 days after left carotid artery (LCA) ligation. After 10 minutes of circulation, the mice were anesthetized with zoletil and rompun, and pressure perfused with saline containing heparin (10 U/ml) via the left ventricle after severing the inferior vena cava to remove unbound phage. The LCA was then isolated. The carotid lumen was rapidly flushed using a 29-gauge insulin syringe with 200 μL of phage elution buffer (0.2M Glycine-HCl, pH 2.2) for detachment of bound phages from endothelium and subsequently neutralized with 30 μL of 1 M Tris-HCl (pH 9.1) using a 29-gauge insulin syringe into a microfuge tube. Eluted phages from LCA were amplified in Escherichia coli ER2738 for reinjection in the next round. Two peptides, CLIRRTSIC and CPRRSHPIC, selectively bound to arterial endothelial cells (ECs) exposed to disturbed flow not only in the partially ligated carotids but also in the lesser curvature and branching point of the aortic arch in mice as well as human pulmonary artery branches. Peptides were conjugated to branched polyethyleniminepolyethylene glycol polymer to generate polyplexes carrying siRNA targeting intercellular adhesion molecule-1 (siICAM-1). In mouse model, CLIRRTSIC polyplexes carrying si-ICAM-1 specifically bound to endothelium in disturbed flow regions, reducing endothelial ICAM-1 expression. Mass spectrometry analysis revealed that non-muscle myosin heavy chain II A (NMHC IIA) is a protein targeted by CLIRRTSIC peptide. Further studies showed that shear stress regulates NMHC IIA expression and localization in ECs. The CLIRRTSIC is a novel peptide that could be used for targeted delivery of therapeutics such as siRNAs to pro-atherogenic endothelium. 3001 CKIMISISC(30) CKMTRSTIC(22) CHIPSIHSC(8) CTHRRSTPC(5) CPLLTKLKC(4) CQMMLIRRC(4) CILRKIRPC(4) CLIRRTSIC(2) CLHPPLTLC(2) CLIRLILQC(2) CMRQRRNRC(2) CNPMTRLIC(2) CNLLLRTQC(1) CSRRPTSIC(1) CRMRKTLRC(1) CQPHHSIIC(1) CSTHIPSHC(1) CILRRRKRC(1) CPRRRLKRC(1) CHQLPIRPC(1) CPIKTTITC(1) CRRQTSTHC(1) CPIRHRPIC(1) CTLHKSRSC(1) CTRIISNTC(1) CKLINTLKC(1) CQMTRSTIC(1) 27 Phage display (in vivo) 3 PMID:27173134 Endothelial cells (ECs) of non-ligated right carotid artery (RCA) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Briefly, 1.0e11 plaque-forming units (pfu) of phages were injected intravenously into mice at 3 days after left carotid artery (LCA) ligation. After 10 minutes of circulation, the mice were anesthetized with zoletil and rompun, and pressure perfused with saline containing heparin (10 U/ml) via the left ventricle after severing the inferior vena cava to remove unbound phage. The unligated right carotid artery (RCA) was then isolated. The carotid lumen was rapidly flushed using a 29-gauge insulin syringe with 200 μL of phage elution buffer (0.2M Glycine-HCl, pH 2.2) for detachment of bound phages from endothelium and subsequently neutralized with 30 μL of 1 M Tris-HCl (pH 9.1) using a 29-gauge insulin syringe into a microfuge tube. Eluted phages from RCA were amplified in Escherichia coli ER2738 for reinjection in the next round. The peptide CLIRRTSIC selectively bound to arterial endothelial cells (ECs) exposed to disturbed flow not only in the partially ligated carotids but also in the lesser curvature and branching point of the aortic arch in mice as well as human pulmonary artery branches. Peptides were conjugated to branched polyethyleniminepolyethylene glycol polymer to generate polyplexes carrying siRNA targeting intercellular adhesion molecule-1 (siICAM-1). In mouse model, CLIRRTSIC polyplexes carrying si-ICAM-1 specifically bound to endothelium in disturbed flow regions, reducing endothelial ICAM-1 expression. Mass spectrometry analysis revealed that non-muscle myosin heavy chain II A (NMHC IIA) is a protein targeted by CLIRRTSIC peptide. Further studies showed that shear stress regulates NMHC IIA expression and localization in ECs. The CLIRRTSIC is a novel peptide that could be used for targeted delivery of therapeutics such as siRNAs to pro-atherogenic endothelium. 3002 NHVHRMHATPAY(3)[0.434 ± 0.067, 0.272 ± 0.041] THAAHMGYPSWW(1)[0.452 ± 0.062, 0.286 ± 0.013] TKNMLSLPVGPG(1)[0.312 ± 0.023, 0.252 ± 0.030] WPHNWWPHFKVK(1)[0.312 ± 0.013, 0.329 ± 0.087] 4 Phage display (subtractive panning) 3 PMID:27168498 Serum from mice with traumatic brain injury (TBI) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Each phage clone was evaluated using ELISA against serum samples from uninjured and injured mice. The absorbance was read at 450 nm. Data shown were reproduced from the graph in the published paper. Data in the first column and the second column represent affinity values of phages for serum samples from injured and uninjured mice, respectively. To eliminate clones that bind serum from control mice (without traumatic brain injury), the library was preincubated with a 96 well plate (Nunc Immuno Plate, Maxisorp surface) coated with pooled serum from control mice diluted 1:100 in TBS (100 μl). 3003 STYTSVS(2) SYYTSVS(1) SHYTSVS(1) SYYTSVS(1) VTDTSVS(1) ATYTSVS(1) TYTSVS(1) VTYTSVS(1) SHLYYVS(1) NSPWLMT(1) PNMQSQS(1) KLRWNQT(1) YRNTPLS(1) APPPSPT(1) PPARHRC(1) APAQAKS(1) YLCGSR(1) LMIRQPD(1) TSVS 18 Phage display (subtractive panning) 3 PMID:23375098 Helix 69 of Escherichia coli 23S rRNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Before the positive selection, the phage solution (10 μL of the original library in 100 μL buffer A, 2.0e11 pfu, in which pfu is plaque forming unit) was pipetted into a well having no immobilized target RNA and incubated for 1 h at RT with gentle shaking for prescreening of the library. 3004 TPSHPLT(4) TSPHPLT(1) TRSHPLT(1) TPRHPLT(1) HPL 4 Phage display (common panning) 3 PMID:23375098 Streptavidin Biotin Not determined. Not determined. Ph.D.-7 phage display library (X7) 3005 STYTSVS(11) NQVANHQ(5) STYLYCA(1) PDYTSVS(1) YYAAESL(1) IRLPNHQ(1) TSVS 6 Phage display (subtractive panning) 4 PMID:23375098 Helix 69 of Escherichia coli 23S rRNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Before the positive selection, the phage solution (10 μL of the original library in 100 μL buffer A, 2.0e11 pfu, in which pfu is plaque forming unit) was pipetted into a well having no immobilized target RNA and incubated for 1 h at RT with gentle shaking for prescreening of the library. One selected sequence, NQVANHQ, was shown through a bead assay to interact with helix 69. Electrospray ionization mass spectroscopy revealed an apparent dissociation constant for the amidated peptide and helix 69 in the low micromolar range. This value is comparable to that of aminoglycoside antibiotics binding to the A site of 16S rRNA or helix 69. Helix 69 variants (human) and unrelated RNAs (helix 31 or A site of 16S rRNA) showed two to four fold lower affinity for NQVANHQ-NH2. These results suggest that the peptide has desirable features for development as a lead compound for novel antimicrobials. 3006 STYTSVS(8) NQVANHQ(4) TSVS 2 Phage display (subtractive panning, competitive panning) 4 PMID:23375098 Helix 69 of Escherichia coli 23S rRNA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Before the positive selection, the phage solution (10 μL of the original library in 100 μL buffer A, 2.0e11 pfu, in which pfu is plaque forming unit) was pipetted into a well having no immobilized target RNA and incubated for 1 h at RT with gentle shaking for prescreening of the library. For round 4, specific elution was carried out with 30 pmol of H69 RNA in TBST buffer. One selected sequence, NQVANHQ, was shown through a bead assay to interact with helix 69. Electrospray ionization mass spectroscopy revealed an apparent dissociation constant for the amidated peptide and helix 69 in the low micromolar range. This value is comparable to that of aminoglycoside antibiotics binding to the A site of 16S rRNA or helix 69. Helix 69 variants (human) and unrelated RNAs (helix 31 or A site of 16S rRNA) showed two to four fold lower affinity for NQVANHQ-NH2. These results suggest that the peptide has desirable features for development as a lead compound for novel antimicrobials. 3007 CAKATCPAC(26) CSWQIGGNC(20) CTVRTSADC(12) CIGNSNTLC(10) CIKVGKLQC(2) CNWGDRILC(2) CTNANHYFC(2) CGWSGSLVC(1) CLYANSPFC(1) CSISSLTDC(1) CNWGDRIRC(1) CNWGDRIEC(1) CRSANIFTC(1) CFNMFSRFC(1) CISAWPTRC(1) CSISSLTHC(1) CFLNPTTTC(1) CKNLTRLAC(1) CMARYMSAC(1) CIKVGKVQC(1) 20 Phage display (in vivo) 4 PMID:25366491 Human adenocarcinoma lung cancer cell line A549 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) For in vivo biopanning, a mouse bearing A549-derived xenograft tumor was injected with 1.0e9 pfu of the random library in TBS buffer 200 μl via the tail vein. After 15 min of biocirculation, the mouse was anesthetized by i.p. injection of zoletil (40 mg/kg) plus rompun (10 mg/kg) mixture and perfused with 50 mL of Tris-buffered saline (TBS; 50 mM Tris–HCl (pH 7.6), 150 mM NaCl) to remove non-specific and unbound phages. In subsequent rounds of biopanning to fourth rounds, different concentrations of detergent in the TBS buffer were used to wash out effectively non-specific bound phages. The increasing concentrations of Tween-20 were as follows: 0.1 % for second round, 0.3 % for third round and 0.5 % for fourth round in the TBS buffer. Then tumor tissues were resected from the mouse and homogenized in 1 mL of cold TBS containing 1× protease inhibitor cocktail (Tech and Innovation Co., Ltd, Seoul, Korea) on ice. Bound phages were subsequently eluted from the homogenate by 0.2 M glycine–HCl (pH 2.2) and the mixture was neutralized with 1 M Tris–HCl (pH 9.1). Novel peptides were categorized into four groups according to a sequence-homology phylogenicity, and in vivo tumor-targeting capacity of these peptides was validated by whole body imaging with Cy5.5-labeled phages in various cancer types. The result revealed that novel peptides accumulated only in adenocarcinoma lung cancer cell-derived xenograft tissue. For further confirmation of the specific targeting ability, in vitro cell-binding assay and immunohistochemistry in vivo tumor tissue were performed with a selected peptide (G9, LYANSPF). The peptide was found to bind intensely to lung cancer cells both in vitro and in vivo, which was efficiently compromised with unlabeled phages in an in vitro competition assay. In conclusion, the peptides specifically targeting human lung cancer were discovered in this study, which is warranted to provide substantive feasibilities for drug delivery and imaging in terms of a novel targeted therapeutics and diagnostics. 3008 DPQYTRFHQHPQ[1.443, 0.266] DQHYTRFHQHFR[1.432, 0.271] QLGHYDRFHKHP[1.421, 0.277] SMNPTWLRFHPH[1.247, 0.260] YPTFERFHTHTP[1.215, 0.250] SSMLNRFHIHTL[1.231, 0.250] IPYTRFHDHQYT[1.383, 0.260] STSASYTRFHSH[1.394, 0.250] Y-x-R-F-H-x-H 8 Phage display (common panning) 3 PMID:27191594 Anti-VP3 monoclonal antibody 4A6 Capsid protein VP3 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Each phage clone was evaluated using ELISA against mAb 4A6 and mAb anti-porcine IFN-c (Sigma, StLouis, MO, USA) as a negative control. The absorbance was read at 450 nm. Data shown were reproduced from the graph in the published paper. Data in the first column and the second column represent affinity values of phages for mAb 4A6 and mAb anti-porcine IFN-c, respectively. Phage clone peptides displayed a consensus sequence of YxRFHxH that mimicked the sequence 82Y/FNRFHCH88, which corresponded to amino acid residues 82 to 88 of VP3 protein of WPVs. 3009 SYQYHTLTYTEL[1.404 ± 0.065, 0.147 ± 0.038] GMAGVQYHPLHL[1.511 ± 0.073, 0.147 ± 0.038] VNALYSVQYQPL[1.576 ± 0.078, 0.147 ± 0.043] SYLSVQYEPLLT[1.598 ± 0.086, 0.138 ± 0.043] ASVDYYTLTDLR[1.451 ± 0.065, 0.142 ± 0.038] LEKFNMVQGQHT[1.365 ± 0.060, 0.138 ± 0.039] AYVTYTPLYTTA[1.348 ± 0.065, 0.142 ± 0.038] SLQYSYYYEEYY[1.343 ± 0.065, 0.138 ± 0.043] SVQYHPL 8 Phage display (common panning) 3 PMID:23185456 Anti-gp90 monoclonal antibody A9E8 Envelope glycoprotein gp90 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Each phage clone was evaluated using ELISA against mAb A9E8 and an anti-porcine IFN-c mAb (negative control). The absorbance was read at 490 nm. Data shown were reproduced from the graph in the published paper. Data in the first column and the second column represent affinity values of phages for mAb A9E8 and an anti-porcine IFN-c mAb, respectively. The mAb A9E8 recognized phages displaying peptides with the consensus motif SVQYHPL. Amino acid sequence of the motif exactly matched 213SVQYHPL219 of the gp90. 3010 SDSLPTPTSNT[1.443 ± 0.098, 0.268 ± 0.060] SMLTAPPTSRD[1.247 ± 0.093, 0.264 ± 0.064] HSYPTPRTSSG[1.357 ± 0.102, 0.264 ± 0.064] VDEYGTALTS[1.417 ± 0.090, 0.264 ± 0.064] SHTQDPAPTSNV[1.319 ± 0.098, 0.264 ± 0.059] TQMLPAPDLPS[1.468 ± 0.094, 0.264 ± 0.064] ATSTQPATSNT[1.506 ± 0.102, 0.264 ± 0.064] NTLPANVPSN[1.204 ± 0.098, 0.268 ± 0.064] LPAPTS 8 Phage display (common panning) 3 PMID:25706372 Anti-VP1 monoclonal antibody 2D10 Capsid protein VP1 Not determined. Not determined. Ph.D.-12 phage display library (X12) Each phage clone was evaluated using ELISA against mAb 2D10 and an anti-porcine IFN-c mAb (negative control). The absorbance was read at 450 nm. Data shown were reproduced from the graph in the published paper. Data in the first column and the second column represent affinity values of phages for mAb 2D10 and an anti-porcine IFN-c mAb, respectively. The mAb 2D10 recognized phages displaying peptides with the consensus motif LPAPTS. Sequence alignment showed that the epitope 173LPAPTS178 is highly conserved among the duck hepatitis A genotype 1 virus (DHAV-1) genotypes. 3011 CYWGGTEGAC(9) CFGAHGVFFC(8) 2 Phage display (common panning) 4 PMID:27226142 Low affinity immunoglobulin gamma Fc region receptor III, IgG Fc receptor III Not determined. Not determined. Not determined. CX8C fUSE5 phage display library The two novel CD16 ligands identified negatively regulate bacterial killing and inflammation. 3012 VVPWAGL(12.5%) WTVHTLS(25%) HMFPWRQ(12.5%) QSWLPSL(8.4%) GYKDFSA(16.6%) RILITIP(12.5%) WASKVAR(8.4%) 7 Phage display (common panning) 3 PMID:26896129 Hemagglutinin-esterase, HE Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 3013 TPARHIY(16.6%) TLLRVDN(11.1%) GSGNAFM(27.7%) MPLGFKA(16.6%) NERALTL(16.6%) VQPIPAT(11.1%) 6 Phage display (common panning) 3 PMID:26896129 Fusion protein Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 3014 TRRAFGSSFRLS SAARTLEVHSSS TSAGDIVVLISL LSRSYCRPPAPC EAESRVYRPGTS SYFPFTYKITRN FPPIFAEATARS QTISLPLDSPKR SGVYKVAYDWQH 9 Phage display (subtractive panning) 3 PMID:26970402 C-terminal half of aryl hydrocarbon receptor (AhR) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Biopanning was performed as follows: HisTag Dynabeads, BSA, and Ph.D.-12 library were incubated in biopanning buffer using a rotating device to preclear the phages. The precleared solution was added to a new tube containing 6His-AHR-N△515 and HisTag Dynabeads. The biopanning process was performed three times to generate the final phage solution. A secondary screening was performed as follows: a LI-COR nitrocellulose membrane (Lincoln, NE) was used for membrane lift to create a mirror image of the individual colonies on the membrane. This membrane was subjected to denaturation/neutralization, and then the membrane was dried and blocked. The blocked membrane was incubated with the IRDye800-conjugated 6HisAHRN△515. After incubation, the membrane was washed, followed by LI-COR Odyssey analysis to detect the colonies bound with AHR-ND515 according to the near IR fluorescence intensity. Eight 12mer peptides, in the form of GFP fusion, suppressed the ligand-dependent transcription of six AHR target genes (cyp1a1, cyp1a2, cyp1b1, ugt1a1, nqo1 and ahrr) in different patterns in Hep3B cells, whereas the AHR antagonist CH223191 suppressed all these target genes similarly. Three of the 12mer peptides (EAESRVYRPGTS, FPPIFAEATARS and TRRAFGSSFRLS) suppressed the 3MC-induced, CYP1A1-dependent EROD activity and the ROS production caused by benzo[a]pyrene. These 12mer peptides suppressed the AHR function synergistically with CH-223191. 3015 SNQLPQQ 1 Phage display (common panning) 4 PMID:25761931 Neural stem/progenitor cell Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Screening of the phage library was performed by the BRASIL method, which allows the separation of cell-bound from unbound phage in one step. The peptide with the highest affinity, SNQLPQQ, was expressed in conjunction with a bispecific adaptor molecule. To verify the targeting potential of the specific peptide, green fluorescent protein-expressing Ad vectors were coupled with the adaptor molecule and injected into the subventricular region of adult mice by stereotaxic surgery. An efficient and selective transduction of NSPCs in the SVZ was achieved, whereas hippocampal NSPCs were negative. 3016 RVTGMMLWD(27) VTGMSLRPD(16) PVTGMAALA(4) RLTGMMLWD(3) RRLVTGMSLY(3) RVTGMSFII(3) RLIGMMLWDP(1) VLTPMMLAW(1) VVTPMMLAW(1) VSGMSLRPD(1) VTGMSLVQG(1) VSGMSLVHG(1) VTGMSALAP(1) RVTGMSISV(1) GRVTPMSVS(1) GSVTPMAIV(1) GAVTPMAIV(1) MVVTGMALV(1) VRVTPMILI(1) VTPMAVRLE(1) CTGLVRMN(1) RVTGMSLV 21 Phage display (common panning) 7 PMID:27216028 Granzyme-like I Not determined. Not determined. Not determined. X9 T7 phage display library 3017 CSSPIGRHC(8) CTMSNLKGC(1) CNNVLSQMC(1) 3 Phage display (in vivo) 3 PMID:27221614 Human non-muscle-invasive bladder cancer cell line BIU-87 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Tumor-bearing mice were intravenously injected with phage peptide library via the tail vein. The isolated phage display peptide (CSSPIGRHC, named NYZL1) was tested in vitro for its binding specificity and affinity. Accumulation into xenograft tumors in a nude mouse model was analyzed with FITC-labeled NYZL1. NYZL1, with strong tumor-homing ability, was identified by in vivo phage library selection in the bladder cancer model. The NYZL1 phage and synthetic FITC-labeled NYZL1 peptides bound to tumor tissues and cells, but were hardly detected in normal control organs. Notably, accumulation of FITC-NYZL1 in bladder tumor cells was time-dependent. Biodistribution studies of xenografts of BIU-87 cells showed accumulation of injected FITC-NYZL1 in the tumors, and the bound peptide could not be removed by perfusion after 24 h. The mouse model of bladder tumor showed increased fluorescence intensity in the tumor-bearing bladder in comparison with normal bladder tissues after 4-6 h. NYZL1 may represent a lead peptide structure applicable in the development of optical molecular imaging. 3018 FPWTEPSYKQGD FPWTEPLYKYGE FPWTEPSYKYGD MPWKESAWLEKI MPWTEPNYLLTQ IPWKESACLAKI APWLEGSYKTVY KQFSALPFNFYT SEFPRSWDMETN 9 Phage display (common panning) 5 PMID:27209258 Human prostate cancer cell line PC3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3019 CNLSSSWIC(5) CPSSQRFFC(2) CLNSLSRHC(1) CSLNSDLFC(1) CTSATSNSC(1) CVQSNPLSC(1) CSNNLLHNC(1) CRPLSFGHC(1) CSSNLPLSC(1) CSSHSWRLC(1) CLPALTRSC(1) CTQLARSAC(1) CLAPQSHRC(1) CSPNAPNSC(1) CHGPVSRSC(1) CSMLSVAAC(1) CSQWSPHRC(1) CQATLHLGC(1) CPSVPHAHC(1) 19 Phage display (common panning) 3 PMID:27313992 Anti-GRA2 monoclonal antibody C3C5 Dense granule protein 2, Protein GRA 2 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 3020 YTHNEKPSSDTH(5)[0.945 ± 0.051] YTVPDNHKYSAH(3)[0.900 ± 0.064] THPWQVSTINFK(3)[0.956 ± 0.055] IHKDKNAPSLVP(2)[0.957 ± 0.048] DVFPPRSHADEL(1)[0.957 ± 0.056] 5 Phage display (subtractive panning) 4 DOI:10.1007/s10989-013-9367-7 Gastric cancer cell line SGC-7901 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Each phage clone was evaluated using ELISA against SGC-7901 cells. Phage clone displaying irrelevant peptide (PIRPs) and PBS were used as negative controls. Plates were read by using a microplate reader (BioRad Model 550, Hercules, CA) at 450 nm. Data shown were reproduced from the graph in the published paper. The affinity value of phages displaying PIRPs for SGC-7901 cells was 0.032. And the affinity value of PBS for SGC-7901 cells was 0.021. The 12-mer phage peptide library was preincubated with HEK293 cells to remove any HEK293 cells-binding phage before being added to the SGC-7901 cells-coated cultures. 3021 YFPYSHTSPRQP[1.304, 0.183] AYKYTSALPAEA[0.970, 0.200] SLTLMNSPLGAS[1.161, 0.161] MLTLSLNPTNSA[0.517, 0.200] 4 Phage display (common panning) 3 PMID:27246497 Protein EaeB Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding affinity of the positive phage clones and control vcsM13 to EspB and 6× His was determined by ELISA. Absorbance was measured at 450 nm. Phage clones were considered to possess high affinity for EspB if their OD values were two-fold greater than those of the control phage and peptide. Data shown were reproduced from the graph in the published paper. The binding affinity of the control vcsM13 to EspB and 6× His was 0.230 and 0.183, respectively. Data in the first column and the second column represent affinity of phages to EspB and 6× His, respectively. 3022 NPMIRRQ(5)[1.349 ± 0.130] MRMTIIN(3)[1.458 ± 0.160] LRLRNTR(1)[1.686 ± 0.172] SHLRHRI(1)[1.797 ± 0.142] KLPLTTK(1)[1.241 ± 0.187] PIKTNRK(1)[1.134 ± 0.041] KPTIPTK(1)[1.499 ± 0.081] 7 Phage display (subtractive panning) 5 PMID:27313733 Human ovarian cancer cell line HO-8910 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding affinity of the positive phage clones to HO-8910 cells was determined by ELISA. The M13K07 phage and phosphate-buffered (PBS) saline were used as negative controls. Absorbance was measured at 490 nm. Data shown were reproduced from the graph in the published paper. The binding affinity of M13K07 phage and PBS to HO-8910 cells was 0.041 ± 0.015 and 0.061 ± 0.031, respectively. The phage peptide library was preincubated with Chinese hamster ovary cell line (CHO) to remove any CHO cells-binding phage before being added to the HO-8910 cells-coated tubes. Immunofluorescence and immunohistochemical assays revealed that the phage clone with the sequence of NPMIRRQ was able to bind to ovarian cancer cells and tissues, and not those of cervical cancer. The peptide may be a potential agent for the diagnosis of ovarian cancer. 3023 SGVYKVAYDWQH(5/34) SSLELSRSSAAS(2/34) QTFSRPDWTMYL(2/34) DPDDTHIMRLLY(1/34) XKSTASMFTTAR(1/34) GSAARWPFSVTL(1/34) LTNTSQLQTRIG(1/34) SQDIRTWNGTRS(1/34) IEINATRAGTNL(1/34) SMRASYPMPTFI(1/34) GLHTSATNLYLH(1/34) FPGLFEMVESLN(1/34) VNMVPLGKNVVQ(1/34) IDAHFGLRLVND(1/34) GSGASYQVWRPM(1/34) MHYGTYTVGYKQ(1/34) VLKEASHLPYSG(1/34) GDYGDGFRLLYI(1/34) VAGLKEQAELDY(1/34) TSWNMGLTYAGQ(1/34) VLTNIARGEYMR(1/34) GQAGGEWILHPL(1/34) AASSGVTVTLPT(1/34) GESVSRIDVGSA(1/34) TTVTGASFSAAY(1/34) IEASFYDAPRGG(1/34) KASGSPSGFWPS(1/34) GSWGLNDSHSYR(1/34) 28 Phage display (common panning) 2 PMID:27486695 Human blood-brain barrier (BBB) cellular model Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) From the panning experiment of a 12-mer library, the SGVYKVAYDWQH (SGV) peptide sequence was selected and internalisation studies suggested that SGV internalises through a clathrin-mediated mechanism and that it increases the uptake of a cargo in endothelial cells. 3024 SGVYKVAYDWQH(2/31) KLNSLDSGMRLV(1/31) AFNPSTPHLRLI(1/31) GLTGDLSLNTAT(1/31) ETLSSYVKKPGH(1/31) AIYMDTGPLAGR(1/31) SLPVGSSSDGWV(1/31) LPLSYNDKARAE(1/31) RVSIESHHLDHY(1/31) DGAVSYLQLRVT(1/31) GLHTSATNLYLH(1/31) FIPFDPMSMRWE(1/31) LDQLPCCWSKTG(1/31) WVNDTNSPLVPR(1/31) MLAPRQTEHGRI(1/31) LMKSPQPLHSSR(1/31) MHPNAGHGSLMR(1/31) GQAPSVPFFASI(1/31) NERNHEIMMAQA(1/31) HEMYSSFGALTV(1/31) NMANYSAISSRW(1/31) STPIFAEATARS(1/31) HSADINVGSRTR(1/31) DEAAYGLKLPGK(1/31) DLGRASSILPSG(1/31) GLFGNEARTAST(1/31) YTNDHSRPKLVP(1/31) SVNSDHTSMRSH(1/31) AVMPRDHVLNAT(1/31) LHRQDNYLAASX(1/31) 30 Phage display (common panning) 3 PMID:27486695 Human blood-brain barrier (BBB) cellular model Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) From the panning experiment of a 12-mer library, the SGVYKVAYDWQH (SGV) peptide sequence was selected and internalisation studies suggested that SGV internalises through a clathrin-mediated mechanism and that it increases the uptake of a cargo in endothelial cells. 3025 LAKPHTENHY(4/32)[1.02 ± 0.11] QNPIQKIIY(1/32)[0.81] LAKPHTENHLLT(1/32)[1.16] 3 Phage display (subtractive panning) 6 PMID:27491899 The envelope protein Ampho4070A Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of the positive phage clones to the envelope protein Ampho4070A expressed by retrovirus like particles were measured by phage ELISA. The OD450 values were measured and data shown were reproduced from the Fig. 1b in the reference. In order to eliminate unspecific binders several depletion steps were incorporated in the panning campaigns using the VLP null particles as well as non-coated plates. 3026 NHLSTPVWSITG(10/20) NSWIQAPDTKSI(6/20) DQFVHDVKGTKH(4/20) 3 Phage display (common panning) 4 PMID:27296960 The cell surface of multidrug resistant Listeria monocytogenes Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Peptide DQFVHDVKGTKH exhibited features common to antimicrobial peptides (AMPs), and was rich in Asp, His and Lys residues. Peptide NSWIQAPDTKSI inhibited bacterial growth in vitro, without any hemolytic or cytotoxic effects on eukaryotic cells. 3027 GYFDVVLGGFGP(5/18)[0.675 ± 0.1] GFYDFILAPLSQ(3/18)[0.782 ± 0.1] GLFDYLQENYRT(1/18)[0.695 ± 0.1] GYYDERQLSDNA(1/18)[1.203 ± 0.1] STHRIEDPWLTR(1/18)[1.109 ± 0.1] GWFDERSFWSNT(1/18)[0.746 ± 0.1] VHWDFRQWWQPS(1/18)[0.62 ± 0.1] GFLDEWVPTGHT(1/18)[1.012 ± 0.1] SGVYKVAYDWQH(1/18)[0.553 ± 0.1] GVYDESSATRLM(1/18)[0.443 ± 0.1] GLLAQEPNNPDY(1/18)[1.095 ± 0.1] GIKDLYDAYYLI(1/18)[0.953 ± 0.1] 12 Phage display (common panning) 3 PMID:27421965 Deinagkistrodon acutus venom Anti-Deinagkistrodon acutus venom polyclonal antibody Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of the positive phage clones to the Deinagkistrodon acutus venom were measured by phage ELISA. The A450 values were measured and data shown were from the Table 1 in the reference. 3028 CKTPNGHLC CPVPQTSEC CDTNGRVSC CEIPGKVVC CPTASNTSC 5 Phage display (subtractive panning) 4 PMID:27548261 Mammary adenocarcinoma murine 4T1 cell line Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Before biopanning,the negative control 3T3 murine fibroblast cell line was used to eliminate non-specific peptides. 3029 EVQSSKFPAHVS 1 Phage display (subtractive panning) 4 PMID:27548261 Mammary adenocarcinoma murine 4T1 cell line Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After biopanning,the negative control 3T3 murine fibroblast cell line was used to eliminate non-specific peptides. 3030 CAQK(22%) CAQSSGDCC(15%) CAQNSGDCC(4%) CAQKSGDCC(3%) 4 Phage display (in vivo) 1 PMID:27351915 Traumatic brain injury (TBI) Not determined. Not determined. Not determined. CX7C T7 phage display library A T7 phage library that displays on the phage surface 9-amino acid cyclic peptides with the general composition of CX7C was intravenously injected 6 h after PBI. Phage was harvested 30 min after injection from the injury site and the corresponding contralateral hemisphere. 3031 CAQK(83%) CRDLAGDTC(6%) CLGGSEKSC(6%) CRDKTPARC(5%) 4 Phage display (in vivo) 2 PMID:27351915 Traumatic brain injury (TBI) Not determined. Not determined. Not determined. CX7C T7 phage display library A T7 phage library that displays on the phage surface 9-amino acid cyclic peptides with the general composition of CX7C was intravenously injected 6 h after PBI. Phage was harvested 30 min after injection from the injury site and the corresponding contralateral hemisphere. 3032 CSSPPRSAGTCG AEGEFAHSGTADVK CYRLQELALGCG AEGEFTAARSNHQP CTQRRSLVFSCG AEGEFPHNTQPQES AEGEFIALHSQPPL AEGEFILHKGFVRW AEGEFQYSSQQGRL AEGEFQQVHFKKHE AEGEFTTRHSVATW CGPSKPPLQYCG CVSPLQSQISCG CLQSKRPCPHCG CLPVRSQGHSCG AEGEFRPGGRGGST AEGEFQNLQIRHRT AEGEFKIHNSPPTM AEGEFQNLQIRHRT AEGEFTQTKRNMSW CPSTLRQPCSCG AEGEFLPKHKQNGG AEGEFFAASCTRQL CSIFQYTALQCG AEGEFTAARSNHQP AEGEFAQNSHLYPQ AEGEFLSTMARSRS CQTAVPSFMVCG CIALQQVCGLCG CPTRVIQPKPCG 30 Phage display (common panning) 2 PMID:27510219 Anti-Salmonella Typhimurium polyclonal antibody IgY Not determined. Not determined. Not determined. pVIII-9aa and pVIII-9aa.Cys phage display library pool 3033 WVCRWYSLWCQE(22) FSCWWLWQPCLW(3) EFCTVLFWLCLG(1) SGCRLWWMICLE(1) SRCRVWAVWCFW(1) WACTWYSLWCDN(1) 6 Phage display (common panning) 3 PMID:27474962 Staphylococcus aureus biofilm Not determined. Not determined. Not determined. X2CX6CX2 M13 phage display library 3034 TDGRRYSSGAMR(19) TRYSSGSTRVSG(6) DVGRRFSSASTR(3) LAGWTDWRPLAG(1) NRYSSGSTRVSG(1) TDGRRYSLFNMR(1) TRFSSRSVGVWW(1) 7 Phage display (common panning) 3 PMID:27474962 Staphylococcus aureus biofilm Not determined. Not determined. Not determined. X12 M13 phage display library 3035 ALRMPTMKTFIP AQAKNLRMPFTK EFLSKVRLPMAK KVMMPLEKNWGY TVKMPSDKISRH SIAEVRLPGAKL LALDSHPFYIPS SWMPHPRWSPQH 8 Phage display (subtractive panning) 3 PMID:27034429 Lambda fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM023 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3036 QNLTFISLPGNI NIFTPLPGNLME RFLRVCLVICRT YVSLPGNASSIR YSVVSLPGNLEK VQGTVSLPGNMT YFPSPNHQISRL 7 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM025 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3037 YPSLFRPTAFNN TYPIHGALSKGG SAMAGASAMSTM ALANMSPVSAMV 4 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM031 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3038 YDRLLYQSTLRY HTNQTYLTTLKY TLYSTTLTYSTP YSTTLMYSNITP YSTTLSWGEKPH RILLFIASTKIY SQTVIYNTTMGY QNTPLFRTTYFY 8 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM040 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3039 EGSILYYTSKTW SDTNWYRATLHY EPTYYSPTLYFG ELEKAYKTTLSY HTNQTYLTTLKY TNYAYTTTLVYV WPSYPNQPHQK 7 Phage display (subtractive panning) 3 PMID:27034429 Lambda fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM040 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3040 MFDVRPPGNTFK MHQPWDVPPMRW YEQSWDLPPLGL THFLWDVAPTGR ASWDIAPVNSTS VEPWDISPTNIF HVLKPVLSGKAA 7 Phage display (subtractive panning) 3 PMID:27034429 Lambda fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM041 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3041 SPSTVAGVTLLD SPNLTKGLSMWP SPSAVVGTNLLR GPSTISGLSMTT 4 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM043 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3042 DVNTRRGIDLLK LPTPVQGSLTKN TAVPQGQLTKTF HSAGLTQGRLDK VLPNTSSGRLLM TPCAANGKMLMA SPSVITGHQLAT MPVPPYSGGQLM 8 Phage display (subtractive panning) 3 PMID:27034429 Lambda fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM043 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3043 NTVDGDDIYLTP LVLESHPNRHGQ GVTSPGSHWFTV GHLHERQFWFTV WIEPMKGPATWS VPAWITTMMSNK FMYPGETMVLAD NEWSPMALGAPP 8 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM045 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3044 KAIEEPKAMMYL NAKVREEPVWHI IGKTVDEPKSHW SQGDRMLHSPLL GQDTNLHKIFNT NEWLLHNIPFRS VPSLKEGEKIWW 7 Phage display (subtractive panning) 3 PMID:27034429 Lambda fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM045 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3045 YRPDEFWSPRKS EAYAPDTFWLTH IYWPPERHWQPI IEMWPTERAWRG DVFRYNPEYFWR 5 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM046 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3046 QSMHIGSSSVSG FVAEHVGNRYVM 2 Phage display (subtractive panning) 3 PMID:27034429 Lambda fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM046 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3047 TNSMPPDAYTE TPENAYSSNTPT IPPENAYGTTRM ISLRHPENAYNK LGARVPENAYNR QLPPESAYNIVL STLHMPENAYGQ PPANYYPSDIMY 8 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM047 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3048 MGFERNPPRVLS GSFNPERDLGIP NAEYPRNPERDA FPWHLVNKPSHR VKNPVPPPWSFY GTEFGDKLTERT MVPKTHGDYHTL 7 Phage display (subtractive panning) 3 PMID:27034429 Lambda fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM047 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3049 HGVSFERYNLSL NAFHGASMTTAQ YTTTSTSQRPVQ VDCPVKWHALCT NNAFSDASRSVT HALSNSSTSMDT 6 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM051 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3050 MVEDDLSSPRYM GVELPLHLSSPR HPFDLSSPRQRY 3 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM051 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3051 GLSRWVEVLALQ GLLRYIDDLTSH NLRCTLFRAWYN VHWDFRQWWQPS 4 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM035 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3052 SPTPSSSMYTLR WPSSLLTDYPPR DPYQVIWYSHDA MLEPDPYQMTWA 4 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM077 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3053 THMWVWDVSPEL NGAPLWDMPPHH HFAPWDILPTSK DKLWDIKPLITA AWDWDMPPLRHV SFHWDLRPYSKL SPKPLWDLRPLH 7 Phage display (subtractive panning) 3 PMID:27034429 Kappa fractions mixture of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM023 and MM031 and MM041 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screenings were performed after twofold negative selection on polyclonal immunoglobulin G. 3054 RHLLLNQ(10/15) MTGNHNS(2/15) SFSLKNW(1/15) GERHYPQ(1/15) 4 Phage display (subtractive panning) 3 PMID:27479451 Peptide nanotubes (PNTs) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The eluted phages were incubated with a SiO2 chip for 1 h, to conduct negative screening of the phages which bind only to this substrate. 3055 CSSAGSLFC 1 Phage display (common panning) 4 PMID:27556860 Anti-human VEGF monoclonal antibody Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 3056 MHPNAGHGSLMR(3) QGGIPLSRTTFV(1) SLFGCSGICLKA(1) AAFPSLNLRTQP(1) KPGDTAMHYFPP(1) TSTNGKAAILVV(1) DRSHNFWFESGE(1) WDEQARTYPLRS(1) GDGNSVLKPGNW(1) 9 Phage display (subtractive panning) 3 PMID:27336166 Cellulose of paper Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Toner was initially treated with the phage library to eliminate non-specific peptides. 3057 MHPNAGHGSLMR(3) GVILVLLGLCSF(1) SPVAVRSNVFVQ(1) KVPVGVLPLSHS(1) KASGSPSGFWPS(1) MPVKHMPKAHWI(1) DFSMDTHINYRR(1) TPQSFWQKGSLV(1) 8 Phage display (subtractive panning) 4 PMID:27336166 Cellulose of paper Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Toner was initially treated with the phage library to eliminate non-specific peptides. 3058 LPVNGDAELWHS(9) SGVYKVAYDWQH(2) SQDIRTWNGTRS(1) VPTSQAGSGTVT(1) 4 Phage display (subtractive panning) 3 PMID:27336166 The printed toner of standard office laser printers Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Cellulose was initially treated with the phage library to eliminate non-specific peptides. 3059 LPVNGDAELWHS(9) SGVYKVAYDWQH(2) 2 Phage display (subtractive panning) 4 PMID:27336166 The printed toner of standard office laser printers Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Cellulose was initially treated with the phage library to eliminate non-specific peptides. 3060 NRPDSAQFWLHH(6) SMAGEQISWALI(3) VPSIDVSTVSYP(1) GTIQPYPFSWGY(1) WTPTIRTQFVPA(1) QPGHAILAQHPT(1) SARLSMHEMTYL(1) YVNSHSILGYTG(1) TGHLYPTRMEIQ(1) NFTLMNAKTFRW(1) HSDNHYRPADKL(1) EELEWTRKAPMV(1) TDNTMYDKQFQK(1) YPTDWLWHGHNK(1) NASTLPLQKYPT(2) 15 Phage display (subtractive panning) 5 PMID:27582001 Recombinant human prostate-specific membrane antigen (PSMA) extracellular domain (ECD) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phages from the previous round were incubated with PC-3 cells (PSMA negative) for one hour to remove non-specific bound phages. 3061 NRPDSAQFWLHH(10) YPTDWLWHGHNK(4) HSDNHYRPADKL(3) QPGHAILAQHPT(3) TGHLYPTRMEIQ(2) SMAGEQISWALI(1) GTIQPYPFSWGY(1) YVNSHSILGYTG(1) KLSTTHDRLMLN(1) TVPGDSSPPRLD(1) 10 Phage display (subtractive panning) 5 PMID:27582001 Recombinant human prostate-specific membrane antigen (PSMA) extracellular domain (ECD) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phages from the previous round were incubated with PC-3 cells (PSMA negative) for one hour to remove non-specific bound phages. The phages from the fourth round were used for in vivo biopanning against xenograft LNCaP tumor. 3062 CKNPTTGTC(14) CGVHSQSSC(9) CSSSPYAWC(6) CWSSFRDEC(4) CNTPMQRSC(3) CSLRGHDLC(2) CTKHTLSIC(2) CKSTPTNGC(1) CPPSKYHLC(1) CPQHFKTFC(1) CITTPGFLC(1) CGEASSPRC(1) 12 Phage display (common panning) 3 PMID:27579674 Anti-carbohydrate larval antigen (CarLA) mAb PAB1 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 3063 SVSVGMKPSPRP(19)[1.034] TMGFTAPRFPHY(12)[0.955] SVCVGMKPSPRH(2)[1.127] YVSVGMKPSPRP(1)[1.044] SMGFTAPRFPHY(1)[0.826] SMGLGVPRFPHH(1)[0.532] SVGLGTKRIPHH(1)[0.522] 7 Phage display (subtractive panning) 5 PMID:27609463 Tuberculosis (TB)-positive sera Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The collected phages were used for two rounds of negative biopanning severally targeting TB-negative serum and the blank plate respectively to eliminate non-specific phages. 3064 IQSPHFF[0.88] 1 Phage display (common panning) 3 PMID:27639393 Tyrosinase Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 3065 TLHPAAD 1 Phage display (common panning) PMID:27555439 Colon cancer cell line SW480 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) According to the results of phage ELISA, phage clone displaying TLHPAAD peptide (SW-TUP clone) showed a significantly higher binding to empty wells than SW480-containing wells. SW-TUP was also found to have a higher binding to empty wells compared with M13KE phage or the whole Ph.D.™-7 library. However, the binding of SW-TUP to SW480-containing wells indicated a slight but insignificant increase compared with both types of control phage clones. These oservations exhibited the TLHPAAD-driven binding of SW-TUP phage to polystyrene. 3066 AEHNPRS AGTTGSL ALAPPVA APAVYLQ APDSGPM APTLVTL ATDASGM DEATSLK DQPAGHL DSARPAS GDLPLAM GLLNPSS IPRDPTT ISPQTGS KVSQPYT MGTYLHG MYTYPIS QHHNPMA QLPHVTQ QPTMQAS RALGPSR SSQPFWS STAHPLP SVTVPPP TAHPILP TASELSR TMQRGAA TPPSLPP TPTISER TSFERSQ VGRAAPL YTTSVRE 32 Phage display (in vivo) 1 PMID:24832468 Glioma-initiating cell, GIC Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) To adapt phage display into anin vivo tumor, we selected a flank tumor model to permit the recovery of a large number relatively pure tumor cells. Bulk glioblastoma cells were implanted into the flanks of BALB/c nu/nu mice. The flank tumor grew until it reached 1 cm in diameter, then the phage display library was injected via the tail vein of the mouse and allowed to circulate and bind to cellular targets for 24 h. 3067 RCQYPLCS(14/40) SCKTVFCY(3/40) RCLRSHCG(3/40) RCPRFSCW(2/40) SCFRPTCP(2/40) RCQYSPCH(1/40) 6 Phage display (common panning) 3 PMID: Fluorescent phosphor LaPO4:Ce3+,Tb3+ (LAP) Not determined. Not determined. Not determined. LX-4 phage display library (XCX4CX) A phage clone containing the surface peptide loop RCQYPLCS was found to bind specifically to LAP. Specificity and affinity of the identified phage bound peptide was confirmed by using binding and competition assays, immunofluorescence assays and zeta potential measurements. Binding and immunofluorescence assays identified the peptide’s affinity for the fluorescent phosphor components CAT and BAM. No affinity was found for other fluorescent phosphor components such as YOX. The binding specificity of the RCQYPLCS peptide loop was improved 3-51-fold by using alanine scanning mutagenesis. The identification of peptides with high specificity and affinity for special components in the fluorescent phosphor in CFLs provides a potentially new strategic approach to rare earth recycling. 3068 VPGWSQAFMALA(21) DTDWVRMRDSAR(9) DMTYMERRDSAR(2) NHVPQHTPIHLR(2) AHVPQHHAMTGR(2) VPGWSQTFMPLA(1) GHVTQHSQRGYS(1) GHVPQHARLLGL(1) THVPEHLLKPRP(1) GHVQQHDVHSIR(1) YHVPEHAVRLGP(1) KHVLQHQTSMTM(1) KHVVQHEYAPNA(1) HHTDEHWLFAKK(1) LSHRTHDRIGFT(1) YPPQERTVHKYV(1) DAYTQLRDRMRQ(1) IDGYPGSPRLPW(1) WHWTWPADVGWV(1) H-V-P-[GQ]-[WH]-S-Q, D-T-x(4)-R-D 19 Phage display (subtractive panning) 3 PMID:27659529 Anti-EGFR monoclonal antibody panitumumab Epidermal growth factor receptor (EC:2.7.10.1) Not determined. Not determined. Ph.D.-12 phage display library (X12) In each round of selection, phage clones were preabsorbed by human IgG immobilized on rProtein A sepharose beads to remove unspecific phages, followed by positive selection with panitumumab counterpart. To enhance the immune responses, we generated recombinant proteins of P19 (DTDWVRMRDSAR) or P26 (VPGWSQAFMALA) fused to a heat-shock cognate protein 70 (Hsc70), and evaluated the efficacy of Hsc70-P19 and Hsc70-P26 as vaccines in vivo. Immunization with Hsc70-P19 or Hsc70-P26 fusion protein stimulated the immune system to produce specific antibodies against peptides as well as EGFR. Moreover, antibodies elicited against mimotopes could induce antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and inhibit the proliferation of EGFR-overexpressing A431 cells. Treatment with Hsc70-P19 and Hsc70-P26 significantly reduced tumor growth in BALB/c transplantable lung cancer models. Although there was no sequence homology between the phage-derived peptides and EGFR by alignments, both peptides mimic the conformational structure of EGFR binding to panitumumab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hage display (common panning) 3 PMID:25989088 Human β-Ⅻa Not determined. Not determined. Not determined. TATA-cyclized CXnCXnC phage display library (CXnCXnC) Inhibition assay The inhibitory activity of bicyclic peptides was determined by incubation with proteases and quantification of their residual activity with a fluorogenic substrate. The inhibitory constant Ki (nM) was calculated and shown. NT denotes not tested. 3070 KCSFREAVCN(13)[NT] ACGRDGFLCQIMRMKCG(2)[>100] ACEMEGWWCTEPVVTRG(2)[NT] VCELWYIVACV(2)[NT] WCAPVFCWWQHCP(2)[NT] YCGELWCWYWMCG(2)[NT] ACKQQGFLCSIATMKCG(1)[>100] YCFPGCAKPCV(1)[NT] WCQDFWCSPVWCL(1)[NT] WCGPWYCMWEGCQ(1)[NT] TCYKFFCGDAVCT(1)[NT] GCWAWFCDYQGCI(1)[NT] WCYGAFCFGQTCF(1)[NT] WCYGGFCWLHWCQ(1)[NT] WCPFMVCWWGTCQ(1)[NT] KCFQGVCYDMWCP(1)[NT] QCCWWFCWDSDCT(1)[NT] MCCTSGCEFQDCY(1)[NT] QCWEPWCCLWLCA(1)[NT] CCLWGFCDQDWCW(1)[NT] CCQELWCYLSFCI(1)[NT] ACNVGDFLCQIQLMKCG(1)[NT] ACQAGEFICAVSANKCG(1)[NT] ACGAQGFLCATSEMKCG(1)[NT] ACENYDFRCAVQVLKCG(1)[NT] ACVKPMFSCFVNQNRCD(1)[NT] ACQPNSGLCWKPVAKCG(1)[NT] ACRTQNPYCQLKQEACG(1)[NT] ACLFMNPYCQVKELMCG(1)[NT] ACQDSVKPCYSAVVLCG(1)[NT] ACRRIQPFCLLLLAVCG(1)[NT] ACQYWDGTCCLSHNSCG(1)[NT] ACDNQCPRCISPWLMCG(1)[NT] 33 Phage display (common panning) 3 PMID:25989089 Anti-ferritin polyclonal antibody Not determined. Not determined. Not determined. TBAB-cyclized CXnCXnC phage display library (CXnCXnC) Inhibition assay The inhibitory activity of bicyclic peptides was determined by incubation with proteases and quantification of their residual activity with a fluorogenic substrate. The inhibitory constant Ki (nM) was calculated and shown. NT denotes not tested. 3071 HPNVPFGVIADG(2)[1.06 ± 0.23] VSDYAFGYWNTL(1)[1.09] GFDIPFGVSLGV(1)[1.29] GSFAMKELYDPF(1)[0.77] QSMASKDLPFGQ(1)[1.31] VVPVDPPFGITW(1)[1.54] DDVPFGRNPAMG(1)[1.45] GNCTLMPPDPPF(1)[1.07] EEPFGTFVDRMA(1)[1.02] EASYPFGYFATA(1)[0.92] ELFDPFGNGLDW(1)[1.25] AISWTDVPFGSK(1)[1.12] TLSSGERAFGSL(1)[1.29] EPAFGLPPMAQK(1)[1.39] SELYKPFGTFRH(1)[1.20] LANNSAEKEYPW(1)[0.90] SALSTGGHDPPF(1)[0.95] ARDYPFGVYHSR(1)[1.12] EASYPFGYFATA(1)[1.31] DPPFGFNHAMY(1)[1.23] SGQATSEVLPPF(1)[1.09] LANNSAEKEYPP(1)[0.92] ERLEIEPSWGF(1)[0.80] NLGGKDPLFGIY(1)[0.76] LANRDPAFGSLS(1)[0.91] E-X-[ED]-P-P-F-G 25 Phage display (common panning) 3 PMID:27824100 Anti-E protein monoclonal antibody (mAb) 3B6 E protein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The OD450nm value was measured in a microplate Reader. The data shown were reproduced from Figure S1. A minimal B-cell epitope (EXE/DPPFG) that mediates binding to a nonneutralizing monoclonal antibody was identified. It is identical to amino acids 374 to 380 (EVEPPFG) of the E protein of duck Tembusu virus (DTMUV). DTMUV-positive duck serum reacted with the epitope, and amino acid substitutions revealed the specific amino acids that are essential for antibody binding. Dot-blot assays of various flavivirus-positive sera indicated that EXE/DPPFG is a cross-reactive epitope in most flaviviruses, including Zika, West Nile, Yellow fever, dengue, and Japanese encephalitis viruses. These findings indicate that the epitope sequence is conserved among many strains of mosquito-borne flavivirus. Protein structure modeling revealed that the epitope is located in domain III of the DTMUV E protein. 3072 TWTCSDVICTAR(3)[3.14 ± 0.05] SCTSPHCFMWLP(4)[3.34 ± 0.04] TGYICDEMFCKL(1)[3.01 ± 0.05] 3 Phage display (subtractive panning) 4 PMID:27832083 Outer membrane protein U, OmpU Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The OD492nm value was measured in a microplate Reader. In the test, M13 phage in peptide library was used as negative control and TBST as blank control. Each clone experiment was performed in triplicate. The OD492nm of M13 phage and TBST is XX and yy, respetively. The data shown were reproduced from Figure 1. An additional incubation step with His tag protein was added in the first round of panning, and the unbound fraction was used for panning with OmpU. Synthetic OmpU-binding peptides (TWTCSDVICTAR designated P1, SCTSPHCFMWLP designated P2, TGYICDEMFCKL designated P3) were measured for their adhesion antagonistic activity and binding affinity via adhesion inhibition test and non-competitive ELISA, respectively. The results showed that after co-incubated with the mixture of rOmpU and P3, visible green fluorescence could be observed on the epithelioma papulosum cyprinidi (EPC) cells surface; while the EPC cells co-incubated with the mixture of rOmpU and P1/P2 exhibited little green fluorescence. The averagead hesion number of V.mimicus 04-14 isolate before and after treatment with peptide was 21.4±1.5, 20.8±0.8(irrelevant peptide), 20.2±0.5 (P3), 5.1±0.7 (P1) and 3.4±0.8 (P2), respectively. There was a significant decrease in the adhesive level of 04-14 isolate treated with P1/P2 compared to the untreated isolate (p<0.01). The affinity constants of P1 and P2 were (6.17±0.19)e8 L/mol and (1.24±0.56)e9 L/mol, respectively. Furthermore, protective effects of P1 and P2 on grass carps challenged with V.mimicus were preliminary detected. It was found there was delayed death of fish in the groups treated with P1/P2, and the survival rate of challenged fish improved with the increase of the dose of adhesion antagonistic peptide. 3073 ELNLPWQRNALV[1.47] SAENDLTLPWTT[1.14] MANAEIDLPWTK[1.23] HPHDLNDLTSPF[1.13] EFWTALSDPWYF[1.29] AHLHDPFTTLSP[1.00] LDFHDLNRPFNN[1.29] THDPLDSPWNFS[0.99] FNDLDLPFGKRA[1.11] SYDLDLPWIARK[1.08] SFLELDPPWTTN[1.16] QHSFLDLPWHLT[1.08] HPHDLNLPTSPF[1.33] HPHDLNLPTSPF[1.22] MANADLNLPWTK[1.37] TSHSWDLNLPSG[1.43] D-L-[DN]-L-P-W-T 16 Phage display (common panning) 3 PMID:27834908 Anti-E protein monoclonal antibody (mAb) 1F3 E protein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The OD450nm value was measured. The data shown were reproduced from Figure 1. One minimal epitope was mapped to 221-LD/NLPW-225 and by using phage display and mutagenesis. Duck Tembusu virus (DTMUV) positive duck sera reacted with the epitope, thus indicating the importance of the minimal amino acids of the epitope for antibody-epitope binding. The performance of the dot blotting assay with the corresponding positive sera indicated that 221-LD/NLPW-225 was a cross-reactive epitope for West Nile virus (WNV), dengue virus (DENV), and Japanese encephalitis virus (JEV) and corresponded to conserved and variable amino acid sequences among these strains. The structure model of the E protein revealed that LD/NLPW was located on domain (D) II, which confirmed that DII might contain a type-specific non-neutralizing epitope. 3074 SRNLSYAEYIQI[1.53] GNYSEYIVGKLV[1.19] SSYANYIQFRNT[1.02] SSYTAYIMARGQ[1.19] NSMSEYINYILT[1.25] VDYSTYISRLTS[1.19] NFMNYAEYVQKK[1.45] VDYSTYISRLTS[1.22] TVHSYEEYTARR[1.01] VSPYAEYWLSQM[1.39] WDYNLYIKYVAR[1.12] VDYATYISRLTS[1.40] Y-A-E-Y-I 12 Phage display (common panning) 3 PMID:27834908 Anti-E protein monoclonal antibody (mAb) 1A5 E protein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The OD450nm value was measured. The data shown were reproduced from Figure 1. One minimal epitope was mapped to 87-YAEYI-91 by using phage display and mutagenesis. Duck Tembusu virus (DTMUV) positive duck sera reacted with the epitope, thus indicating the importance of the minimal amino acids of the epitopes for antibody-epitope binding. The performance of the dot blotting assay with the corresponding positive sera indicated that YAEYI was DTMUV type-specific. The structure model of the E protein revealed that YAEYI was located on domain (D) II, which confirmed that DII might contain a type-specific non-neutralizing epitope. The YAEYI epitope-based antigen demonstrated its diagnostic potential by reacting with high specificity to serum samples obtained from DTMUV-infected ducks. Based on these observations, a YAEYI-based serological test could be used for DTMUV surveillance and could differentiate DTMUV infections from JEV or WNV infections. 3075 CRVDLQGWRRCRR[1.80] CRAWYQNYCALRR[0.61] VPCPYLPLWNCAGK[8.20] 3 Phage display (competitive panning) 2 PMID:27836545 Phospholipid hydroperoxidase glutathione peroxidase (GPX4) mutant, GPX4mu Not determined. Not determined. Not determined. CX7-10C T7 phage display library Surface plasmon resonance (SPR) SPR biosensing experiments were performed on a Biacore T100 instrument (GE healthcare).The dissociation constants KD (μM) were shown. Biotinylated Avi-tagged GPX4mu was immobilized to Dynabeads M280-streptavidin (Invitrogen, CA, USA). After one panning agaist SUMO-tagged GPX4mu, the phage libraries were re-panned against SUMO-tagged GPX4mu in the presence of 50 μM SUMO-tag binding synthetic peptides identified from the first panning. The first GPX4 inhibitory peptide, VPCPYLPLWNCAGK, was identified. By analyzing the X-ray crystal structure of the peptide, we found the peptide binds to near Sec73 of catalytic site. This peptide may contribute to developing GPX4-targeted drugs. 3076 TTFNSFGRVRIE(2) TTFNSFGRVAIE(1) TTFCSFGRVRIE(1) TTFNSFGRVHWE(1) TTFNSFGKVRIE(1) TTYNSFGRVRIE(1) TTEYSFGRTSTL(1) DTFNSFGRVRIE(1) TTFNSFGRVRIQ(1) TNFNDFKRVRGE(1) TTFSSFGRVRIG(1) TTFNSNGRVWIE(1) TTFNSFGRVRGG(1) TTFNSFYRVRIE(1) 14 Phage display (common panning) 8 PMID:27842517 Human breast cancer cell line MDA-MB-231 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3077 PRLNVSP(2) PRLNTSP(1) PTLDVAP(1) PQLNVSP(1) PGAQVSP(1) PRTNVAP(1) PRKTVSP(1) PAMNVSP(1) PQENASP(1) PSLNVSP(1) ARLNVAP(1) TMLMVRP(1) ARLNTQP(1) PMMAVAP(1) 14 Phage display (common panning) 8 PMID:27842517 Human breast cancer cell line MDA-MB-231 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 3078 PRQNQSP QLKNVTP PRLNVAT PRLNVTT PRWAVSP PRLNHSP PQMTAMP MFLNGAP TRLQVSP WRMAHSP 10 Phage display (subtractive panning) 8 PMID:27842517 Human breast cancer cell line MDA-MB-231 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Screening of the phage library was performed by the BRASIL method. In the final round of panning, bound phages from the previous round were incubated with MCF-10-2A cells (non-tumorigenic), which in this case was the aqueous phase containing the phages that did not bind to the control cells. 3079 WWFNSFGRVRIE(4) WAFNSFGRVRIE(1) WWFNKFGKVRIE(1) IWFNSFFRVRIE(1) WWFFSFGRVRIE(1) GWFNSFGRWSWL(1) WWFFSFGRWSWL(1) 7 Phage display (subtractive panning) 8 PMID:27842517 Human breast cancer cell line MDA-MB-231 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Screening of the phage library was performed by the BRASIL method. In the final round of panning, bound phages from the previous round were incubated with MCF-10-2A cells (non-tumorigenic), which in this case was the aqueous phase containing the phages that did not bind to the control cells. 3080 CTDKSKAGFEC(3/100) CSHDSKLWC(3/100) CKDETKYGC(3/100) CKSDAEWFC(3/100) CPKRISKRC(2/100) CIFISKRIC(2/100) CVITNDPDC(2/100) CWVGLDPDC(2/100) CNEDGLHIC(2/100) CGLKHNEDC(2/100) CHTYLTSTC(2/100) CFLTSDEAC(2/100) CTDRWQKIC(2/100) CASWQKITC(2/100) CPITHWRVC(2/100) CDCGATDGC(2/100) CHHTPAANC(2/100) CRTRFKGCC(2/100) CVSHNDPDC(1/100) CVFDDNGRC(1/100) CRENDNGSC(1/100) CWYTGNGNC(1/100) CRGQRNGWC(1/100) CGQRYEPLC(1/100) CCGVSDTEC(1/100) CCGANQEYC(1/100) CSAKSTYPC(1/100) CCPAIWFTC(1/100) CIVSEWRDC(1/100) CPTISEKNC(1/100) CREFYLSDC(1/100) CSGHPEYLC(1/100) CSGHPEYLC(1/100) CGADEEKLC(1/100) CPEDDFLGC(1/100) CPEDDFGLC(1/100) CVNQNDRMC(1/100) CDTSDQNRC(1/100) CHTEVGCNC(1/100) CMTCPPWFC(1/100) CYPCSGPEC(1/100) CHLCSTVGC(1/100) CPGSATCMC(1/100) CLEFTSAGC(1/100) 44 Phage display (subtractive panning) 3 PMID:27845058 IgGs of horse immunized by the crude venom of the Hemiscorpius lepturus Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The phage-displayed random peptide library was first screened with non-immunized horse IgGs. The unbounded phage (in the supernatant) was screened with the IgG of the immunized sample. 3081 FVRVHTRSSWRVP[0.26] GVWYTDIRMRDWM[0.032] 2 Phage display (common panning) 6 PMID:27856253 BTB domain of B cell lymphoma 6(5-129) BCL-6 corepressors BcoR, SMRT, and N-CoR 1R2B(196), 1R2B(196), X13 T7 phage display library ELISA These peptides exhibited BCL6-selective inhibition in a peptide concentration dependent manner, and the IC50 values (nM) were estimated. 3082 SGADGLVPRQRA[0.73 ± 0.02] VTEADGLVPRNL[1.14 ± 0.03] AHIVSLDDDSTI[0.89 ± 0.02] YSLDDDSTISLN[1.12 ± 0.03] SRDFAPDPTLLM[1.03 ± 0.03] ADGLVPR,SLDDDSTI 5 Phage display (common panning) 4 PMID:27863546 Anti-rVMH-HD polyclonal antibody Not determined. 3BIM(196), 3BIM(196), Ph.D.-12 phage display library (X12) ELISA The binding activities of the positive phage clones to the anti-rVMH-HD antibody were measured by phage ELISA. The OD450 nm values were measured. The data shown were reproduced from the Figure 4 in the reference. 3083 THVSPNQGGLPS(57)[735.2 ± 53.6] TIDNFFSASLRN(13)[NT] THVSPNQGGLRN(1)[NT] 3 Phage display (subtractive panning) 7 PMID:27862961 Hepatocellular carcinoma cell line HepG2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance at 450 nm of the 96-well plate was measured in microtiter plate reader (Thermo Scientific, USA). The apparent Kd value (nM) between biotin-THVSPNQGGLPS and the human recombinant GPC3 protein was calculated and shown. NT denotes not tested. The target is glypican-3 (GPC3) abnormally expressed in hepatocellular carcinoma (HCC). In the first round, PC3 cells were used in negative screening to eliminate non-specific binding clones. Panning procedure was slightly altered in the seventh round by using human recombinant GPC3 protein for competing off the GPC3 binding peptide instead of Tris-HCl. The ability to target GPC3 in vivo is evaluated by intravenous injection of THVSPNQGGLPS (named as GBP) labeled with a near-infrared dye, Cy5.5, into a HCC tumor-bearing mouse model. Significant high tumor accumulation (tumor/muscle ratio: 6.49 ± 0.55) of Cy5.5-GBP in HepG2 tumors is observed compared with that of the low GPC3 expressing prostate cancer cell line, PC3 (tumor/muscle ratio: 1.15 ± 0.32). By targeting GPC3, GBP differentiates tumor tissues from normal liver tissues in patients, suggesting a great clinical translation potency of GBP. Collectively, GBP demonstrates great potential for HCC detection via fluorescent imaging or histological staining. 3084 GEEFSQLHAFEQDIFSC(46) GEAHCKHSEQDICAIAN(4) GVQRSEQDIFTEPEAHD(2) GHQACEVVQTEGFLLHY(2) GQLVCPHVQTEGECVSC(2) GKDSCVSRVQTEGLAVD(1) GRINSGSRQTEGRYGTY(1) GWWFSVLLTQTEGRKQC(1) GAHTCIFQTEGRIHPQF(1) GHRPCVPATEGRQAAHD(1) GTWNSQQTEGRGSDNCA(1) 11 Phage display (competitive panning) 2 PMID:30267550 Serum 13006 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 3085 GGEASIPDIFVQTEGFI(36) GTVVCFIWWPAGYPVDC(6) GSVPSKRFQTEGREQII(4) GGVHSQDYENVQTEGIF(2) GIDGCPITVQTEGQGPD(1) GSGPSSATQVVQTEGQD(1) 6 Phage display (competitive panning) 2 PMID:30267550 Serum 14011 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 3086 GSAPALPVEAPK(14)[743.2 ± 69.5] KMPKENPSSWLS(13)[135.8 ± 8.8] GSAPLLTVDTSP(11)[626.1 ± 40.1] SGVYKVAYDWQH(2)[300.4 ± 30.09] 4 Phage display (common panning) 4 PMID:27721007 Prominin-1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Activities of phages to the CD133 were measured by phage ELISA. The OD values at 450 nm were measured. The data shown were the Kdapp value (apparent dissociation constant, pM), which were obtained from a binding saturation and derived from three independent experiments. 3087 AYPQKFNNNFMS(16/20) DLLAMHWNTSRQ(1/20) IQAATVPHVTES(1/20) KVKHPSSWAYYA(1/20) SNSIDKVNRPIN(1/20) 5 Phage display (common panning) 4 PMID:27133793 TiO2 nanoparticles Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3088 QPTKDSSPPLRV(5)[0.338176 ± 0.002] STTSPPAVPHNN(1)[0.3243] PMPQYKLVVVGS(1)[0.35764] KAPTKETVWPSR(1)[0.3513] 4 Phage display (common panning) 3 PMID:27650372 The LMP1 C-terminus region Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Activities of phages to the LMP1 C-terminus region were measured by phage ELISA. The OD values at 410 nm were measured and data shown were reproduced from the Fig 3 in the reference. 3089 TLWTDTS GLWTNVD GLSTFDV TAFTWNQ 4 Phage display (common panning) 3 PMID:27711976 Pentaerythritol trinitrate hemisuccinate(PETNH) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 3090 LPGRTCREYPDLWWVRCY 1 Phage display (common panning) 5 PMID:27717820 The FGFR1c/KLB complex Fibroblast growth factor 21, FGF-21 Not determined. Not determined. X5CX10CX T7 phage display library (X5CX10CX) 3091 ITPAHMD(1) WDSSY@V(1) SQHTHRH(1) SGSSDQG(1) 4 Phage display (common panning) 1 PMID:27720899 Tumor necrosis factor-α(TNF-α) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Symbol @ denotes an amber stop codon. 3092 AHQQHLR(3) ATPIHWR(2) HLSHQPG(1) NISFQSS(1) 4 Phage display (common panning) 2 PMID:27720899 Tumor necrosis factor-α(TNF-α) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 3093 HAIYPRH(11)[0.51] NLMSRPI(10)[1.47] AHQQHLR(3)[NT] ATPIHWR(2)[NT] STAPRPY(1)[NT] QFPSFTQ(1)[NT] 6 Phage display (common panning) 3 PMID:27720899 Tumor necrosis factor-α(TNF-α) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The absorbance was measured at 450 nm using ELISA reader. In order to calculate the binding constant (Kd), the data were analyzed using Prism program (version 6.01, Graphpad Software Inc.) The specific phage with the sequences of NLMSRPI (P51) and HAIYPRH (P52) have ability to specifically bind to TNF-α with Kd of 1.47 and 0.51 nM, respectively. The irrelevant phage has no significant binding affinity towards TNF-α. In addition, the MTT assay was carried out to assess the ability of the selected P51 and P52 peptides to inhibit TNF-αcytotoxicity on L929 cells either as the peptides displayed by the phage particles or the synthesized heptamer peptides. The results of MTT assay showed that P51 and P52 peptides inhibit TNF-α cytotoxicity with IC50 values of 25.15 ± 2.18 and 7.08 ± 2.24μM, respectively. 3094 NLMSRPI(10)[1.47] IHKPQPN(4)[NT] NMMSVRG(1)[NT] LSNMHSQ(1)[NT] 4 Phage display (common panning) 4 PMID:27720899 Tumor necrosis factor-α(TNF-α) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The absorbance was measured at 450 nm using ELISA reader. In order to calculate the binding constant (Kd), the data were analyzed using Prism program (version 6.01, Graphpad Software Inc.) The specific phage with the sequence of NLMSRPI (P51) have ability to specifically bind to TNF-α with Kd of 1.47 nM. The irrelevant phage has no significant binding affinity towards TNF-α. In addition, the MTT assay was carried out to assess the ability of the selected P51 peptide to inhibit TNF-αcytotoxicity on L929 cells either as the peptides displayed by the phage particles or the synthesized heptamer peptides. The results of MTT assay showed that P51 peptide inhibit TNF-α cytotoxicity with IC50 values of 25.15 ± 2.18 μM. 3095 HAIYPRH(11)[0.51] NLMSRPI(10)[1.47] 2 Phage display (common panning) 5 PMID:27720899 Tumor necrosis factor-α(TNF-α) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The absorbance was measured at 450 nm using ELISA reader. In order to calculate the binding constant (Kd), the data were analyzed using Prism program (version 6.01, Graphpad Software Inc.) The specific phage with the sequences of NLMSRPI (P51) and HAIYPRH (P52) have ability to specifically bind to TNF-α with Kd of 1.47 and 0.51 nM, respectively. The irrelevant phage has no significant binding affinity towards TNF-α. In addition, the MTT assay was carried out to assess the ability of the selected P51 and P52 peptides to inhibit TNF-αcytotoxicity on L929 cells either as the peptides displayed by the phage particles or the synthesized heptamer peptides. The results of MTT assay showed that P51 and P52 peptides inhibit TNF-α cytotoxicity with IC50 values of 25.15 ± 2.18 and 7.08 ± 2.24μM, respectively. 3096 CNTGSPYEC CQNPNQKFC CLKLGEKWC 3 Phage display (common panning) 3 PMID:27735110 The heteroternary complex with methyl viologen and cucurbit[8]uril(MV·CB[8]) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) From the selected motifs, an epitope consisting of three amino acids (QKF) was extrapolated and incorporated into a solvent-exposed loop of a protein domain; the protein exhibited micromolar binding affinity for the MV·CB[8] complex, matching that of the cyclic peptide. By achieving selective CB[8]-mediated of a small molecule to a recombinant protein scaffold we pave the way to biomedical applications of this simple ternary system. 3097 FPWYKWRLPDVS(3) GASWLGSQWNGT(1) HLAGVSMDVSTT(1) FAETHRGFHFSF(1) HTSSLWHLFRST(1) NLLASIGPTMRI(1) MPRANASGEPHT(1) DQFVHDVKGTKH(1) 8 Phage display (common panning) 4 PMID:27371890 The acid–alkali treated titanium Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3098 IGNSNTL(18)[0.887±0.037] LKLGEKW(4)[0.864±0.018] LPWQIHN(4)[0.649±0.014] CGNSNTL(4)[0.723±0.025] NTGSPYE(2)[0.425±0.037] NMQITKG(2)[0.335±0.016] PTVLQLT(1)[0.684±0.007] STEHTRS(1)[0.501±0.009] PTNQHHL(1)[NT] VKSTQYT(1)[0.760±0.012] MPVGAFK(1)[NT] DELHKAR(1)[0.637±0.021] ELGTVQS(1)[0.356±0.009] FGAFTPH(1)[NT] FHELTSK(1)[0.434±0.035] 15 Phage display (common panning) 4 PMID:27749918 The fifth fragment of plasminogen Plasmin light chain B Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Affinities of the selected phage clones to Kringle 5 were measured by phage ELISA. The A410 values were measured and data shown were reproduced from the Fig. 2 in the reference. 3099 PWWWGRNV(6) NLWGFWFP(2) ESPLSLVA(1) WWGIWMQE(1) 4 Phage display (common panning) 2 PMID:27756316 Coxsackievirus A9, CAV9 Not determined. Not determined. Not determined. CX8C phage display library 3100 LWWQIWDG(4) PWWWGRNV(3) WIWAWRSS(1) LSWWSRKW(1) WWAIWMQE(1) 5 Phage display (common panning) 2 PMID:27756316 Coxsackievirus A9 lacking the RGD motif (CV-A9-RGDdel) Not determined. Not determined. Not determined. CX8C phage display library 3101 LPSAGRGVCYEA QHLNSILLVTK YPHGYVAFGRW 3 Phage display (competitive panning) 3 PMID:27766741 Anti-LPS single-domain antibody fragment VHH lipopolysaccharide, LPS Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of phage clones to the VHH were measured by phage ELISA. The OD values at 490 nm were measured and data shown were reproduced from the Fig. 2A in the reference. Bound phages were eluted with 100μl elution buffer. In order to compete with the immobilized VHH on the plate, the elution buffer was added VHH in a concentration of 100μg/ml. 3102 EMFTPPSMIERL 1 Phage display (in vivo) 2 PMID:27768975 Ocular tissues Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The Ph.D.-12 library was topically applied into the conjunctival sac or injected into the retrobulbar space of male Sprague-Dawley rats. One hour after administration, aqueous humor was extracted, the eyes were enucleated, and the iris, vitreous humor, retina and choroid were dissociated after washing. A novel dodecapeptide with the sequence of EMFTPPSMIERL, named CC12, was identified with the ability to penetrate the ocular barrier in a noninvasive (via conjunctival sac instillation) or minimally invasive (via retrobulbar injection) manner. KV11, an antiangiogenesis peptide previously demonstrated to inhibit pathological neovascularization in the retina, was then used as a model antiangiogenesis cargo for CC12. It was found that conjugation of KV11 peptide with CC12 peptide facilitated the delivery of KV11 to the retina, resulting in significant inhibition of retinal neovascularization development via topical application without tissue toxicity. 3103 CRGATPMSC(5) CTHLRHASC(4) CMSTGLSSC(3) CGGGPLYMC(2) CSQLPWYSC(1) CPNSTHRNC(1) CPTVKQKWC(1) CNMFDARAC(1) CNPSKEKTC(1) CLMTSQFRC(1) CTRGDRIPC(1) CYENTRHTC(1) CNTGSPYEC(1) CDKVESRVC(1) CKNLSHEPC(1) CQQLTHADC(1) CPTTVGIKC(1) CLKDRIPLC(1) CPLQNSPTC(1) CSIKSKPSC(1) CNNKFLANC(1) CRMNSLLDC(1) CVPSRMEKC(1) CSSDTDKTC(1) CTPFLGRLC(1) CPSWYKNEC(1) CDESKVQTC(1) CSLASAATC(1) CIATSKPSC(1) CTQTGDMAC(1) CAEFTVAGC(1) CLGPGGFSC(1) CEHTANSLC(1) CLQKTYPQC(1) CLYSTKTMC(1) CDAPMKYMC(1) CSSTYLNLC(1) CLNTNTMSC(1) CGNNGNNTC(1) CDELHNAAC(1) 40 Phage display (common panning) 3/4 PMID:27802020 Tetragonal Barium Titanate (BaTiO3 or BT) Nanocrystals Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) After the third round of selection, 50 random colonies were picked out separately for DNA sequencing A short BaTiO3-binding/nucleating peptide, CRGATPMSC (named RS), was identified. We found that the resultant phages could not only bind with the presynthesized BaTiO3 crystals but also induce the nucleation of uniform tetragonal BaTiO3 nanocrystals at room temperature and without the use of toxic reagents to form one-dimensional polycrystalline BaTiO3 nanowires. This approach enables the green synthesis of BaTiO3 polycrystalline nanowires with potential applications in bioimaging and biosensingfields. 3104 CLGTTPFFC(4)[1.52, 16.9] CLYHPWNNC(3)[1.41, 20.1] CTGTTPFYC(3)[1.64, 12.6] CRGSMPFWC(3)[1.38, 18.4] CMARYMSAC(2)[1.59, 13.2] 5 Phage display (common panning) 3 PMID:27874102 Anti-OPs monoclonal antibody mAb3C9 O,O-dimethyl organophosphorus pesticides (O,O-dimethyl OPs) Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The binding activities of phage clones to the mAb3C9 were measured by phage ELISA. The absorbance was recorded at 450 nm. The affinity data in the first column are the maximal absorbance, while the affinity data in the second column are the IC50 (ng/mL). 3105 CPHPQRPAC[0.21] CKIMKASPC[1.00] CASACPPHC[1.33] CGLHKVHKC[0.83] CHEQKSLPC[0.67] CKSSSVLWC[1.22] CLPGKHGHC[3.41] CTSHPRPSC[4.15] CSVTTHLLC[2.01] SKIVSTPSC[2.20] CMLNSKHYC[2.23] CFHLPSTYC[2.01] 12 Phage display (common panning) 2-4 PMID:277290620 Interleukin-7 receptor subunit alpha, IL-7RA Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The binding activities of phage clones to the IL-7Rα were measured by phage ELISA. The OD values at 405 nm were measured. The data shown are the apparent dissociation constant values (Kd, nM). The phage library was pre-panned against the Dynabeads free of protein but blocked with BSA, Fc/IgG-coated Dynabeads and fibronectin coated well before selection agaist IL-7Rα-coated Dynabeads® Protein A or G. 3106 LWWQIWDG(4) PWWWGRNV(3) WIWAWRSS(1) LSWWSRKW(1) WWAIWMQE(1) 5 Phage display (common panning) 2 PMID:27756316 Coxsackievirus A9 lacking the RGD motif (CV-A9-RGDdel) Not determined. Not determined. Not determined. CX8C phage display library 3107 KFPDIDSISSLW(3) STDSIITNSQRL(2) FEDSIVSTRETF(1) YYDDTASIRSSR(1) VNLSDLSLHYPS(1) SGVYKVAYDWQH(1) HTEDDTASITTS(1) DRSSDSIVSWRG(1) AYAGSTGLFERG(1) NFSPSNVPGDKF(1) HVDTGSDKKLDH(1) SDSDSIRTYMNI(1) RSEGEVLSPETL(1) DSISSIVTSQAF(1) MDGLDSIYTSSR(1) GQIIQDFDWVQN(1) SSTDSIMSSYIG(1) ACCDIDSIKSSV(1) 18 Phage display (common panning) 3 PMID:28072528 Anti-Dengue virus (DENV) polyclonal antibody Triosephosphate isomerase (TIM) Not determined. Not determined. Ph.D.-12 phage display library (X12) 3108 RGWGNGCGLF GDQHQVGNET MDFNEMILLT MPLPWTSGAT LPWTSGATTE MGSQEGAMHT SQEGAMHTAL QEGAMHTALT LTGATEIQNS MKKEVSETQH TEDGQGKAHN KGSSIGKMFE MNSKNTSMSF MLWKQIANEL MTVVVGDIIG MMSAAVKDER MAGPISQHNH MVSAGSGKVD YKKGSSIGKMFE LKYSWKTGKAK 20 Phage display (common panning) 2 PMID:28152075 Anti-Dengue virus (DENV) polyclonal antibody Dengue virus (DENV) Not determined. Not determined. Bacteriophage MS2 VLP DENV-3 antigen fragment library ELISA We utilized a pathogen-specific antigen fragment library displayed on bacteriophage MS2-VLPs in combination with deep sequence-coupled biopanning. 3109 RDYHPRDHTATWGGG(1)[0.47857±0.01259] ISSFGNPEFSTGGG(2)[0.03014±0.00254] NYPWMAGTQSMGGG(3) [0.02168±0.00255] GNNPLHVHHDKRGGG(4)[0.34423±0.03388] SHTFVNEHTPPSGGG(5)[0.03148±0.00254] WLDDQTMRNLDSGGG(6)[0.03777±0.00496] SPLRAVAFSGAQGGG(7)[0.03643±0.00375] GADTSKPPRFVTGGG(8)[0.02518±0.02264] VCSPCGPVPPAKGGG(9)[0.04527±0.00121] HMYYPGGDGRFAGGG(10)[0.04648±0.01513] 10 Phage display (subtractive panning) 3 PMID:28186164 NIST RM8670 (NISTmAb) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Absorbance was read at 450 nm. The Absorbance values shown were regenerated from Figure 2 in the published paper. Phage display panning was performed in a subtractive manner. The library was initially depleted with control non-aggregated NISTmAb using three iterative cycles.The unbound phage was then screened on cross-linked NISTmAb aggregates, again using three iterative cycles of phage binding, elution, and amplification. Peptides RDYHPRDHTATWGGG and GNNPLHVHHDKRGGG possess preferential binding to NISTmAb aggregates. 3110 WBRMPAYTAKYP WBRMPMETAKPL KNPTSTPAMYAN KPPQBTSAPYLP GBRNRANTSKNP GHWHSPHKLTRN RxPxxTxK 6 Phage display (common panning) 5 PMID:28220894 GST-N1-Src SH3 domain Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) By screening a peptide phage display library, we discovered a novel ligand (WHRMPAYTAKYP, PDN1) that targets the unique SH3 domain of N1-Src and inhibits N1-Src in cells. In cultured neurons, PDN1 fused to a fluorescent protein inhibited neurite outgrowth, an effect that was mimicked by shRNA targeting theN1-Src microexon. PDN1 also inhibited L1-CAM-dependent neurite elongation in cerebellar granuleneurons, a pathway previously shown to be disrupted in Src−/− mice. PDN1 therefore represents anovel tool for distinguishing the functions of N1-Src and C-Src in neurons and is a starting point for the development of a small molecule inhibitor of N1-Src. 3111 EWWWW 1 Phage display (common panning) 2 PMID:28315327 Kelch-like ECH-associated protein 1 Nuclear factor erythroid 2-related factor 2 Not determined. Not determined. X4 T7 phage display library Biotinylated Avi-tagged Keap1 Kelch domain was immobilized to Dynabeads M280-streptavidin (Invitrogen) in PBS (045e29795, Wako) containing 0.5% BSA. After washing the beads using PBS containing 0.1% Tween 20 (PBST), the beads were incubated with the phage libraries for 1 h and were washed with PBST. The bound phages were eluted with 0.5% SDS and were incubated with E. coli BLT5615 cells (Merck Millipore) in log-phase growth for phage amplification. After bacteriolysis, the phages were recovered from the culture supernatant by centrifugation and PEG-precipitation according to manufacturer's T7 protocol. The recovered phages were suspended in PBS and were used for the next round of panning. We identified a tetrapeptide, EWWW, showing moderate binding affinity, which inhibits the interaction between Nrf2 and Keap1. The tetrapeptide does not include an ETGE motif, which is a commonly found consensus sequence in known peptidic inhibitors. In addition to affinity parameters, IC50, KD, and thermodynamic parameters, the crystal structure of the complex was determined to elucidate the binding conformation. The binding interactions resemble those of known small-molecule inhibitors as opposed to those of substrates and peptidic inhibitors. 3112 ANKNLLFKIRYSTAR 1 Phage display (common panning) 5 PMID:28254693 Toll-like receptor-4 (TLR4) and myeloid differentiation factor 2 (MD2) complex Not determined. Not determined. Not determined. X15 fUSE55 phage display library 3113 LDCFRWGWRMWC YEIRCWWRWCYT 2 Phage display (common panning) 5 PMID:28254693 Toll-like receptor-4 (TLR4) and myeloid differentiation factor 2 (MD2) complex Not determined. Not determined. Not determined. X12 pHEN2 phage display library 3114 HINWPILPRLWV SPDHASRDWRSR TQLDDHKRSHHA ISRPTPSVNPLM LTQGTSDMTRHL SWKPYTWKDTAL FQGSPKTNHGKI GPTFNLQRIPAT QTKSSVYMMSYL WHSNNSHNDSWP ADKPRVDTTTYN IRLPASLLLDPA GPKSNNVGVTYS FWPHKHNLYMST AHWFGGVYNKTM TPLPSPIDITYQ YALHDGTAHNTW LQASAKTMHGTI SNTYLPKSYLNV AHRYIDAQIDRR SMLRQEFPPTEP QTHHHTFFMKSK YYYADDGAILLN 23 Phage display (common panning) 3 PMID:28184219 Anti-E. rhusiopathiae polyclonal antibodies Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Solid phase panning was applied for the first round of biopanning. Solution-phase panning by the affinity SPA Sepharose capture method was applied for second round biopanning. 3115 CMSLLTATC CDLKSTVKC CNNWLTMPC CNTVRESVC CVSRYNWGC CSHNQPYQC CLQAKPRTC CLSKWTTSC CLNANTSLC CSPNYSRNC CNADKPTEC CERTNSSDC CQPSRDTYC CYGDSNLAC CGQEGMKEC CNAMLRAVC CLPFMIHNC CILSDSGSC CPTVKQKWC CSSATLVYC CGAKWMSQC CRDKALVNC CNTTQSVMC CSTSFDNSC CLALDRRDC CYRPTTDQC CLKTLNPSC CQYRNHADC CSFPKNLDC CFQPKRLDC CSSSGQRLC CYRENHSYC CILNYPESC CQEGTNKAC 34 Phage display (common panning) 3 PMID:28184219 Anti-E. rhusiopathiae polyclonal antibodies Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Solid phase panning was applied for the first round of biopanning. Solution-phase panning by the affinity SPA Sepharose capture method was applied for second round biopanning. 3116 NPVEDYLDYSVI ESYYMNPVEMFV LPTESNPVEDWI NPIESYIASVFS NPVELLLKTSSD NPVETQIVLSLV NPVEKWISMRTM NPVELLLLMGIS NGIERLLEEPVS NPVENWIDPKSI VNPVEYYLDTMR NPVEALLSKFHM NPVESYLANYTS VTNPYESLVQEK SPPEFSGSTVGL MVPEFSGSFPMR RMPPEFMGSLPQ QLFVPEFAGSSP LVPEFTGSTPFR SHGAIEFDGAFP VPEFAGHVPSTA VPEMNGSLIARK HPIVKTYFAQTT APAKTYFGQTTD QMKAWFPQTTYD ATKVMFPQRIYV QPAKTYFNQVTL QAAAKTMFPQNT SPSIDAFETSIF SWVLTATETGSS EMRFPSLSASDT QAPTLDAQETAL TALDAVSTGFSW GAIGSLTADSTS SLGSLSAYETGR TLYRPPLTSAET LIADLNAESTSR SLGAAGARVTTV ASGPLHAGATGL GHKPVLTALSTA VMPAIHAGVTGA VHPTLTVTSKEI ELDVSLRVMPKV SKLAMEIMSGPV SNAVTSSKSPRM QADATVLTKPKT KAVSDASRGTVF DYPKIANFQEYA GGQVRSIHSGPT QHWPTNVDSVTV ETKSDDMLLSNV MTVDRTVRVASK DLLIRDAITDTK AYHVDTVSDAGW VDTINLPQNTIQ VMSVNASTTAAN TLHAPMTIRSGP TNLHRVMTVVNM LRPNAVQTDTLA QMRQLTEYGSEK VLSSTAIKVDSV VHTVHDVFTAFG AKIRMFLDTDYK ASWGPIAIDRVN SQQYALTNSTTN QPQTKSFYPQYV NVVDRVNRTGVV WSALPERTSLPV RPAIVDQVSSSP QWNWRVRSVANV AQLHPTTLVKHK TSYRPPLNVCQD QSHSLFYPHPYG N-P-V-E-(x)3, x-P-E-F--G-S-x(2), K-x(2)-F-P-Q-x-T, L-x-A-x-E-T-x 73 Phage display (common panning) 3 PMID:28167275 Serum antibodies Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) We synthesize identified peptides in the array format and rescreen the serum used for phage panning to validate antibody binding peptides. Finally, we systematically vary the sequence of validated antibody binding peptides to identify those amino acids within the peptides that are crucial for binding "their" antibody species. The resulting immune fingerprints can then be used to trace them back to potential antigens. We investigated the serum of an individual in this pipeline, which led to the identification of 73 antibody fingerprints. Some fingerprints could be traced back to their most likely antigen, for example the immunodominant capsid protein VP1 of enteroviruses, most likely elicited by the ubiquitous poliovirus vaccination. 3117 NVRPRICRVRKWTLCF PAKHLCLLAANRVRRSWQCL PRHKKCFRVLRACIEY SDTCWKFYRGWPRRCRSN HAWCFPGPSLWPRRCRNN KSWCRQGFWPIRCSTT GPWKLSFL GPWNYRSYKFWSKTAK AWPYQNWSFWTSRFSD NIPSIRDCIPIGCLQYAFWR KRACKHSHGIWCTKW 11 Phage display (common panning) 5 PMID:28043792 BDNF/NT-3 growth factors receptor (EC:2.7.10.1) Brain-derived neurotrophic factor, BDNF Not determined. Not determined. T7 phage display library After the phage panning against recombinant Fc-fused TrkB (TrkB-Fc), agonistic phages were directly screened against TrkB-expressing HEK293 cells. Through subsequent screening of the first-hit BM17 peptide (NVRPRICRVRKWTLCF)-derived focus library, we successfully obtained the BM17d99 peptide (KSLPRMCRVRKWRLCF), which had no sequence similarity with BDNF but had TrkB-binding capacity. We then synthesized a dimeric BM17d99 analog peptide that could phosphorylate or activate TrkB by facilitating receptor homodimerization. Treatment of TrkB-expressing HEK293 cells with the dimeric BM17d99 analog peptide significantly induced the phosphorylation of TrkB, suggesting that homodimerization of TrkB was enhanced by the dimeric peptide. 3118 RGDDWA(20)[1.66955 ± 0.85] RGDDWE(12)[1.10951 ± 0.85] NRASNR(4)[2.21998 ± 0.85] KKILH*(4)[NT] 4 Phage display (common panning) 4 PMID:16787913 Kappa fractions of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM077 Fibronectin(FN) Not determined. Not determined. X6 T7 phage display library ELISA The binding activities of phage clones to the Lactoferrin (LF) were measured by phage ELISA. The OD values at 450 nm were measured and data shown were reproduced from the Fig 1B in the reference. Two of the four peptides had the typical cell adhesion motif Arg-Gly-Asp (RGD), suggesting that human lactoferrin (hLF) may interact with RGD-containing proteins. Binding experiments using ELISA and surface plasmon resonance (SPR) analysis demonstrated that hLF bound to human fibronectin (hFN) and vitronectin (hVN) with relatively high affinity. Furthermore, the cell adhesion of Chinese hamster ovary (CHO), HeLa, and RAW cells to hFN or hVN coated on the culture plate was inhibited by hLF in a concentration-dependent manner, indicating that hLF interfered the cell adherent interaction through RGD motif. 3119 DVYWDTMAGSWITLTE ELPVDLEHEFQKMYME NDHQGHIKVTEYEVLG FEDLSPLLSKKSHTSS DFLDLTPLVGNAVMPV WQDTTAWFDNYREDTK SFTMYEPDQQTIVIES QSHIITMWRDLTIIDF HQVENGWWWMPHIDHN VDETVEWHDSNMELIH PVQSNKQYVPEPFVWM LDWKWEQLGSPLWTWN SDLVWDDLTPIF 13 Phage display (subtractive panning) 4 PMID:28028054 Kappa fractions mixture of polyclonal immunoglobulin G(IgG) of multiplemyeloma patient MM023 and MM031 and MM041 Not determined. Not determined. Not determined. X16 and X12 M13 phage display library The phage pool was pre-adsorbed on immobilized non-cognate mATG8 proteins. After the pre-adsorption, non-cognate mATG8 proteins was added to the phage pool and then transferred onto the selection plate. In the initial rounds of selections, input phage pools were additionally pre-adsorbed on immobilized GST alone. 3120 GVSNSENVEGAEGFLVY GKLRCENVEGKIHVAYN GVIQCYVPENVEGRSTA GVATSINENVEGYKPDY GNFMSKNVEGNPIVHFC GEVQCVFVPNVEGGEHD GLNQCARVPNVEGEHVD GDSQSKPKQQTCNVEGY GGAHSQNVEGRCSDKDY GYNQCNVEGQCYLNPCF GYYNSIDEANQYRTPCA GEEVSIDEALEKYRDFI GHKMSIDEAFPWEFFQY GIHWCKNEHKFYIDEAN GLIGSFETWGEIDEANY GYHGSTYESIDEAHFDF GEPLSTDEAITGRCHCF GGNFCIDEAQYQFVGPC GWEMCIDEACPCGHEVN GNDFSQTIDEAEYVYRD 20 Phage display (subtractive panning) 2 PMID:27906504 Serum of soy allergic patients Not determined. Not determined. Not determined. X16 phagemid library 3121 VAKYHGYPWNRRR[12 ± 11] LNRCVAKYHGYPWCRRR-NH2[6 ± 2] CVAKYHGYPWCRRR-NH2[5 ± 2] [VLW]-A-[KRL]-[YFW]-[HM]-G-x(2)-W 3 Phage display (competitive panning) 2 PMID:28039053 Dedicator of cytokinesis 2 (DOCK2) Not determined. Not determined. Not determined. CX7-10C T7 phage display library ELISA Biotinylated His-Avi-SUMO-TEV-DOCK2(1192-1622)was captured by the SA, and mixture of synthetic peptide and FLAG-His-TEV-Rac1(1-177) (2.5 nM) was added to the wells. IC50 values (nM) were calculated by Prism 5 (GraphPad Software, CA, USA). His-Avi-SUMOTEV-DOCK2(1192-1622) was immobilized to streptavidin (SA) M280 Dynabeads (Invitrogen, CA, USA) in PBS (045-29795, Wako) containing 0.5% BSA (blocking buffer). After washing in PBS containing 0.1% Tween 20 (PBST), beads were incubated with phage libraries for 1 h, and washed with PBST. Bound phages were competitively eluted by FLAG-His-TEV-Rac1(1-177) (1 mM) and were incubated with Escherichia coli BLT5615 cells (Merck) in logphase growth for phage amplification. These peptides inhibited DOCK2 activity at nanomolar concentrations and were delivered to intracellular compartments by combination with cell-penetrating peptide (CPP). Consequently, one peptide, R4-DCpep-2(V2W/K4R/ox)-NH2 (Ac-RRRRCWARYHGYPWCRRRR-NH2), inhibited migration in human B lymphocyte MINO cell line at IC50 = 120 nM. 3122 IQYRSWIPFSYP LDYRSWAPYATS YRSFDPWYPPVH LSIRSYTSPQWQ LNYRSYLPYETS YRSWxP 5 Phage display (common panning) 3/4 PMID:28035012 Viral protein 1 and 2 (VP1 and VP2) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) After biopanning, phage populations were sequenced and analyzed to identify a consensus peptide motif—YRSWXP. Two 12-mer peptides containing this sequence, NV-O-R5-3 (LDYRSWAPYATS) and NV-O-R5-6 (IQYRSWIPFSYP), were further characterized to evaluate the motif’s functional ability to detect VLPs and virus. Results indicated that these peptides effectively detect GI.1 VLPs in solidphase peptide arrays, ELISAs and dot blots. Further, their specificity for the S-domain of the major capsid protein enables them to detect a wide range of GI and GII norovirus genotypes. Both peptides were able to detect virus in norovirus-positive clinical stool samples. 3123 PPWYMCYPMKLKPDC[1000] RRCPLYISYDPVCRR[51] CMWWREICPVWW[65] 3 Phage display (subtractive panning) PMID:28153726 Recombinant V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (K-Ras) with G12D substitution Not determined. Not determined. Not determined. CX7-10C T7 phage display library Surface plasmon resonance (SPR) To determine binding kinetics of peptides, an evaluation system using SPR was constructed.The KD value to KRas(G12D)in GDP form was shown in Table1. They thoroughly subtracted phages bound to WT K-Ras in the phage panning process. As a result, they successfully discovered K-Ras(G12D)-selective inhibitory peptides. 3124 GASESYL 1 Phage display (common panning) PMID:28208072 Anti-TIM polyclonal antibody Triosephosphate isomerase, TIM Not determined. Not determined. Ph.D.-7 phage display library (X7) A conformational epitope (A71G74S69D75T73F72V67) of TIM was confirmed by the phage display technology. 3125 YSTNWKNLSSLS(7) YSINWKNWLSKY(2) YSTNWKNQLSLS(1) YSMNWKNQLSLS(1) YSMNWKNQSSLY(1) 5 Phage display (common panning) 3 PMID:28272485 Anti-PRRSV monoclonal antibody A1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3126 KSLSRHDHIHHH(6) QVHINHISPSVP(2) KSLSRHDHLLPH(1) QSLSRHDPILHH(1) SFQAGIRSAPEP(1) SFQVGIRSAPEP(1) 6 Phage display (common panning) 3 PMID:28272485 Anti-PRRSV monoclonal antibody A2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Anti-porcine reproductive and respiratory syndrome virus (PRRSV) neutralizing monoclonal antibody A2 recognize the mimotope (KSLSRHDHIHHH), P2, which contains the SRHDHIH motif, which has conserved consensus sequences from amino acid positions 156 to 161 in the N-terminal ectodomain of GP3. The artificial multi-epitope gene, mp2, was designed by combining three repeats of the mimotope P2. The resulting sequence was inserted into the swinepox virus (SPV) genome to construct a recombinant swinepox virus (rSPV-mp2). The rSPV-mp2 was able to stably express the multi-epitope peptide, mP2, in vitro. The rSPV-mp2 immunized pigs exhibited a significantly shorter fever duration compared with the wtSPV treated group (P < 0.05). There was an enhanced humoral and cellular immune response, decreased number of PRRSV genomic copies, and a significant reduction in the gross lung pathology (P < 0.05) was observed following PRRSV infection in rSPV-mp2-immunized animals. 3127 KSLSRHDHIHHH(3) GPRPPMTSTIFP(2) KSFSRHDIHHH(1) KSLSRHDIHQH(1) KSLSRQDIHRH(1) KSVSRQDIHHQ(1) KSSSRHYYLHHQ(1) GPRPPTWSPGQV(1) GPRRRKWYLGQV(1) 9 Phage display (common panning) 3 PMID:28272485 Anti-PRRSV monoclonal antibody A7 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Anti-porcine reproductive and respiratory syndrome virus (PRRSV) neutralizing monoclonal antibody A7 recognize the mimotope (KSLSRHDHIHHH), P2, which contains the SRHDHIH motif, which has conserved consensus sequences from amino acid positions 156 to 161 in the N-terminal ectodomain of GP3. The artificial multi-epitope gene, mp2, was designed by combining three repeats of the mimotope P2. The resulting sequence was inserted into the swinepox virus (SPV) genome to construct a recombinant swinepox virus (rSPV-mp2). The rSPV-mp2 was able to stably express the multi-epitope peptide, mP2, in vitro. The rSPV-mp2 immunized pigs exhibited a significantly shorter fever duration compared with the wtSPV treated group (P < 0.05). There was an enhanced humoral and cellular immune response, decreased number of PRRSV genomic copies, and a significant reduction in the gross lung pathology (P < 0.05) was observed following PRRSV infection in rSPV-mp2-immunized animals. 3128 YAAHRSH(5/50) KFPAINQ(5/50) GVQSPHF(4/50) VPFPSAS(2/50) GIHTLMG(2/50) ATWVSPY(2/50) GSTPNWH(2/50) IDPQTPT(2/50) ADRAWAR(2/50) NHRPHLD(1/50) TNPHLNW(1/50) AFPSPTD(1/50) STSPESA(1/50) ALHGPTP(1/50) APSFTKH(1/50) DFHHASM(1/50) FPPFQTQ(1/50) GHYSAGC(1/50) HSAHENS(1/50) ISPQTPT(1/50) LIAASNN(1/50) LPPSSRM(1/50) NSHSMPF(1/50) QFATQSH(1/50) QIGYQRA(1/50) QSFWFHA(1/50) SVSPISH(1/50) TPLTPHQ(1/50) TSSHAPL(1/50) VALSAPY(1/50) VAPWEKL(1/50) VIIVPPA(1/50) YIPTAMK(1/50) 33 Phage display (common panning) 3 PMID:28252649 Metacaspase, MCA Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Four peptides, NHRPHLD, YAAHRSH, ATWVSPY and AFPSPTD, were selected for their capacity to specifically bind to MCA and interfere with its activity. The peptide with the sequence of YAAHRSH, similar to ecotin-like ISP3 of L. (L.) major, decreases trypsin-like activity of promastigotes under heat shock, and significantly decreases parasite heat shock-induced death. 3129 YEQHKLPSSWPF(12) YQTKLQHVWPFS(4) SPYQTRLPVVWP(3) LSQSKLASPWPL(1) QISQLRLPNNWP(1) FQTRLLHTWPSP(1) EHQVRLSSQWPL(1) TEQMKLLYHWPT(1) QYKIPPLWPMLK(1) YQMKSPTPVWPF(1) HHWFKSLTTKHL(1) YQMKLPLILPPT(1) DFQNKLSSVTLR(1) ISQQPLAPVWPP(1) DAQQLYLSNWRS(1) NQHPPMAHLSAQ(1) YVHHSQLPDASL(1) HAKSQLNPPDIK(1) SAHQVPNAVIKS(1) GQGVPDLHPPLL(1) LPNPQQYTTPSS(1) RYPLESPYWQQT(1) TSPLNIHNGQKL(1) DSTNMETQTLPL(1) DLNTNRTQMVLH(1) HESFWYLPHQSY(1) 26 Phage display (common panning) 5 PMID:18959752 Protein-glutamine gamma-glutamyltransferase K Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The peptides selected as glutamine donor substrate exhibited a marked tendency in primary structure, conforming to the sequence: QxK⁄RψxxxWP (where x and ψ represent non-conserved and hydrophobic amino acids, respectively). Using glutathione S-transferase (GST) fusion proteins of the selected peptides, we identified several sequences as preferred substrates and confirmed that they were isozyme-specific. We generated GST-fused alanine mutants of the most reactive sequence (K5, YEQHKLPSSWPF) to determine the residues that were critical for reactivity. Even in peptide form, K5 appeared to have high and specific reactivity as substrate. In situ analysis of mouse skin sections using fluorescence-conjugated K5 peptide resulted in detection of TGase 1 activity with high sensitivity, but no signal was detected in a TGase 1-null mouse. 3130 WSLSELH(14.3%) DGRYYIN(9.1%) VSRDTPQ(4.5%) MHTVAVQ(4.2%) HLNQQNH(3.5%) APRNVPP(2.4%) WPFHGDN(1.5%) QISAASQ(1.5%) LPYDHLP(1.5%) YPWFIRA(1.4%) 10 Phage display (competitive panning) 1 PMID:26246327 Sera of peanut allergic patient 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3131 VSRDTPQ(33.9%) DGRYYIN(20.2%) WSLSELH(15.8%) HWRLPLH(7.4%) HLNQQNH(4.6%) IWRLPTH(4.2%) MHTVAVQ(2.4%) VMPLDDV(1.4%) SYTDLLR(1.2%) YPWFIRA(0.9%) 10 Phage display (competitive panning) 2 PMID:26246327 Sera of peanut allergic patient 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3132 VSRDTPQ(73.1%) DGRYYIN(9.5%) WSLSELH(8.7%) HLNQQNH(3.4%) HWRLPLH(0.9%) IWRLPTH(0.7%) SYTDLLR(0.7%) VMPLDDV(0.5%) YPWFIRA(0.5%) MHTVAVQ(0.3%) 10 Phage display (competitive panning) 3 PMID:26246327 Sera of peanut allergic patient 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3133 DNYDSTA(59.8%) MLHELLE(3.0%) RVASLAP(2.7%) ENHVHVR(2.6%) NYFKHEA(2.3%) DAIPTSV(1.8%) VSGLLVE(1.8%) ELRNENT(1.8%) NSPELTS(1.1%) NGHSIHT(1.0%) 10 Phage display (common panning) 1 PMID:26246327 Sera of peanut allergic patient 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3134 DSTA(74.0%) VSGLLVE(7.3%) ENHVHVR(5.3%) MLHELLE(4.3%) NSPELTS(2.1%) DAIPTSV(1.7%) SLLGQTP(1.1%) YPWFIRA(0.7%) NYFKHEA(0.5%) ISPLSVP(0.5%) 10 Phage display (competitive panning) 2 PMID:26246327 Sera of peanut allergic patient 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3135 DNYDSTA(64.1%) ENHVHVR(15.5%) VSGLLVE(12.7%) YPWFIRA(4.7%) MLHELLE(2.5%) SLLGQTP(0.1%) NSPELTS(0.1%) SPTTTYD(0.1%) TALRTIS(0.1%) ISPLSVP(0.0%) 10 Phage display (competitive panning) 3 PMID:26246327 Sera of peanut allergic patient 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3136 EVMGRLA(10.4%) STTPHSR(7.5%) MVPPSYD(3.4%) SSIGGQD(2.9%) QLYREFN(2.8%) YLGFDVH(1.8%) SQNTKYI(1.5%) QLIHWRH(1.3%) MVQHYAE(1.2%) HSILSNL(1.2%) 10 Phage display (competitive panning) 1 PMID:26246327 Sera of peanut allergic patient 3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3137 EVMGRLA(72.1%) MVPPSYD(5.8%) QLIHWRH(4.8%) QLYREFN(3.6%) SSNQFHQ(1.5%) HSILSNL(1.5%) YLGFDVH(1.4%) AQYVAVG(0.9%) SSIGGQD(0.8%) ATDEIVP(0.8%) 10 Phage display (common panning) 2 PMID:26246327 Sera of peanut allergic patient 3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3138 EVMGRLA(64.9%) MVPPSYD(8.7%) VKFDKYV(8.5%) AMSHSKQ(5.4%) QLIHWRH(3.1%) YLGFDVH(1.2%) WSLSELH(1.1%) HSILSNL(0.9%) LKFDTPI(0.9%) QLYREFN(0.7%) 10 Phage display (competitive panning) 3 PMID:26246327 Sera of peanut allergic patient 3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3139 SLLGQTP(16.5%) STIVAEM(6.8%) GFTFQPS(5.3%) SAAWNKS(4.7%) TNLSHVP(3.4%) QTWLEMG(3.3%) LLADMHA(2.6%) GTGSQAS(2.0%) LVYVDMH(1.0%) ISYLSVT(0.9%) 10 Phage display (competitive panning) 1 PMID:26246327 Sera of peanut allergic patient 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3140 SLLGQTP(41.9%) SAAWNKS(16.0%) STIVAEM(12.9%) GTGSQAS(6.6%) GFTFQPS(5.9%) QTWLEMG(4.9%) TNLSHVP(2.3%) SIIARPG(1.8%) STPATLI(1.2%) LLADMHA(0.8%) 10 Phage display (competitive panning) 2 PMID:26246327 Sera of peanut allergic patient 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3141 SLLGQTP(50.0%) SAAWNKS(18.9%) STIVAEM(13.1%) GTGSQAS(6.2%) QTWLEMG(4.4%) GFTFQPS(2.5%) KGIYHQV(0.8%) STPATLI(0.7%) SLSSAWW(0.6%) TNLSHVP(0.5%) 10 Phage display (competitive panning) 3 PMID:26246327 Sera of peanut allergic patient 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3142 AMSPLNK(16.2%) MPTWLHH(13.8%) NVRLPYQ(9.2%) NLLDSLH(6.0%) NPTHPIY(3.5%) FASRSDT(2.7%) IENRIYR(2.3%) GVFISYN(2.2%) LSKINSP(2.1%) GNEVMTY(1.6%) 10 Phage display (competitive panning) 1 PMID:26246327 Plasma of peanut allergic patient 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3143 MPTWLHH(29.9%) AMSPLNK(17.5%) GASESYL(10.4%) GVFISYN(5.3%) FASRSDT(4.8%) IENRIYR(4.7%) NVRLPYQ(3.5%) IPNGHFT(3.4%) NLLDSLH(3.3%) NPTHPIY(2.4%) 10 Phage display (competitive panning) 2 PMID:26246327 Plasma of peanut allergic patient 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3144 GASESYL(36.4%) MPTWLHH(26.1%) IENRIYR(13.4%) AMSPLNK(8.6%) FASRSDT(3.6%) NPPWFHT(2.2%) GVFISYN(2.2%) IPNGHFT(1.4%) NVRLPYQ(0.7%) NLLDSLH(0.7%) 10 Phage display (competitive panning) 3 PMID:26246327 Plasma of peanut allergic patient 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3145 LSANHWV(24.5%) VPNIVTQ(7.5%) VTRDSNH(7.2%) SHPLWNS(5.6%) SFNLPNT(4.1%) LPFINSD(2.5%) HTNSAYI(1.7%) GMMSSPP(1.6%) DAIPTSV(1.5%) HAGFVPS(1.4%) 10 Phage display (competitive panning) 1 PMID:26246327 Plasma of peanut allergic patient 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3146 VPNIVTQ(48.7%) LSANHWV(24.3%) SFNLPNT(5.1%) GMMSSPP(4.7%) YVTRTPY(1.8%) HGGVRLY(1.3%) HYIDFRW(1.2%) ATKHYTT(1.0%) AVDPQYG(1.0%) DAIPTSV(0.8%) 10 Phage display (competitive panning) 2 PMID:26246327 Plasma of peanut allergic patient 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3147 VPNIVTQ(75.5%) LSANHWV(5.6%) HGGVRLY(5.0%) SFNLPNT(3.4%) GMMSSPP(3.1%) YVTRTPY(1.5%) VSYGVPM(0.8%) HYIDFRW(0.7%) KGIYHQV(0.3%) AVDPQYG(0.3%) 10 Phage display (competitive panning) 3 PMID:26246327 Plasma of peanut allergic patient 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3148 EFARNSI(21.6%) LDIVDAP(3.6%) ALPLLDA(2.6%) FSTNFGN(2.5%) QHDTVPP(2.1%) HISGRPL(2.0%) VNGHYTI(2.0%) QIGHDGN(1.8%) GQAMNHT(1.8%) DFHPDVL(1.7%) 10 Phage display (competitive panning) 1 PMID:26246327 Sera of peanut-tolerant subject 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3149 EFARNSI(78.7%) QHDTVPP(6.7%) HSTQLPY(3.5%) HYIDFRW(2.2%) FVPHKWY(1.9%) AMSHSKQ(1.2%) FCPDCKP(0.8%) TGKMLAD(0.6%) GAAYIRA(0.5%) NVAHKMF(0.5%) 10 Phage display (competitive panning) 2 PMID:26246327 Sera of peanut-tolerant subject 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3150 EFARNSI(75.7%) HYIDFRW(18.7%) QHDTVPP(1.6%) FVPHKWY(1.1%) YGNSGIV(1.0%) AMSHSKQ(0.8%) FCPDCKP(0.4%) HSTQLPY(0.1%) NVAHKMF(0.1%) TGKMLAD(0.0%) 10 Phage display (competitive panning) 3 PMID:26246327 Sera of peanut-tolerant subject 1 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The ten most frequent peptides were shown. 3151 VSRDTPQ(28.4%) SYTPGGH(16.7%) LIQPITT(7.4%) QLYREFN(4.2%) MDTPDRI(3.5%) WVTDSSW(3.2%) HSPNSIK(2.9%) RFDWSYP(2.9%) AYASDSY(2.7%) YNISVNK(2.6%) 10 Phage display (competitive panning) 1 PMID:26246327 Sera of peanut-tolerant subject 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3152 VSRDTPQ(79.0%) SAEKLLR(6.9%) YNISVNK(3.0%) QLYREFN(2.4%) WVTDSSW(1.7%) HSPNSIK(1.4%) GQSEKHL(1.3%) RFDWSYP(1.2%) LPGNRLL(0.9%) RTWEPYT(0.5%) 10 Phage display (competitive panning) 2 PMID:26246327 Sera of peanut-tolerant subject 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3153 VSRDTPQ(80.3%) SAEKLLR(13.2%) GQSEKHL(3.5%) YNISVNK(1.1%) WVTDSSW(0.8%) QLYREFN(0.4%) LPGNRLL(0.3%) RFDWSYP(0.3%) HSPNSIK(0.0%) RTWEPYT(0.0%) 10 Phage display (competitive panning) 3 PMID:26246327 Sera of peanut-tolerant subject 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3154 SNYHPSI(51.2%) TTQVLEA(3.1%) KMISATE(2.9%) LPQSWAM(1.8%) ATAEWHP(1.7%) APANTFT(1.7%) VPPTEHP(1.3%) LGQLFPQ(1.3%) MIHPRDQ(1.1%) FTQPLYK(1.1%) 10 Phage display (competitive panning) 1 PMID:26246327 Sera of peanut-tolerant subject 3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3155 SNYHPSI(61.0%) KMISATE(6.2%) AERYPDS(5.2%) KMESGTA(3.7%) AKYEAGS(3.2%) TTQVLEA(1.9%) TEQKIEA(1.9%) TITAMQG(1.4%) AERILSV(1.3%) GWETRME(1.2%) 10 Phage display (competitive panning) 2 PMID:26246327 Sera of peanut-tolerant subject 3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3156 AERYPDS(56.8%) SNYHPSI(27.6%) KMISATE(8.0%) ELGTTQT(2.3%) TTQVLEA(1.0%) AKYEAGS(0.7%) GWETRME(0.5%) NDRPHMP(0.3%) ADLMHFT(0.3%) VPPTEHP(0.3%) 10 Phage display (competitive panning) 3 PMID:26246327 Sera of peanut-tolerant subject 3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3157 KSLNSTI(17.2%) VPILNSL(7.1%) SMNETFA(6.6%) WSLSELH(3.7%) NTDTTRE(3.3%) VNQSWDL(2.0%) RDEVYFH(1.9%) NEAPRHA(1.7%) IDRTQFM(1.5%) NTVGTVQ(1.3%) 10 Phage display (competitive panning) 1 PMID:26246327 Sera of peanut-tolerant subject 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3158 VPILNSL(19.8%) WSLSELH(17.0%) ENHVHVR(13.2%) QETRGTY(4.8%) IDNSHTH(4.0%) TTANVRI(3.9%) VNQSWDL(3.1%) HAAWHFI(2.9%) EPAIATP(2.3%) NEAPRHA(2.3%) 10 Phage display (competitive panning) 2 PMID:26246327 Sera of peanut-tolerant subject 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3159 WSLSELH(20.6%) HAAWHFI(19.3%) ENHVHVR(16.7%) VPILNSL(7.6%) VTLPPSS(3.8%) TTANVRI(3.4%) ESKWMPL(3.2%) IDNSHTH(3.2%) QETRGTY(3.0%) GTMHSMK(2.6%) 10 Phage display (competitive panning) 3 PMID:26246327 Sera of peanut-tolerant subject 4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3160 MGQPTVK(58.0%) GPLKTWK(18.6%) GTLASLN(5.4%) KVKTYDM(3.4%) QQLAVDT(2.7%) STLNWWN(1.4%) QTNMRAP(0.4%) ESRVMSR(0.3%) EARMHRA(0.3%) TYPSLWK(0.3%) 10 Phage display (competitive panning) 1 PMID:26246327 Plasma of peanut-tolerant subject 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3161 QQLAVDT(39.8%) MGQPTVK(16.2%) TTQVLEA(8.8%) WEGPQPK(6.9%) GPLKTWK(6.8%) ESRVMSR(4.1%) GTSDKLP(2.3%) KDNTYVM(2.3%) STLNWWN(2.1%) GTLASLN(1.3%) 10 Phage display (competitive panning) 2 PMID:26246327 Plasma of peanut-tolerant subject 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3162 QQLAVDT(64.5%) ESRVMSR(14.4%) WEGPQPK(7.9%) ALGTRMI(6.5%) STLNWWN(2.7%) KDNTYVM(1.0%) TTQVLEA(0.8%) GTSDKLP(0.5%) HSHTLTW(0.4%) YPWFIRA(0.3%) 10 Phage display (competitive panning) 3 PMID:26246327 Plasma of peanut-tolerant subject 2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3163 DVHKLGN(23.8%) ISHTKAE(10.0%) NDIPHLM(9.1%) RIHVHNS(4.4%) AQSLETS(3.5%) SLTMKAN(2.1%) DLNMPIS(2.0%) LYADSVL(1.9%) ITPAYDN(1.8%) SVASTTV(1.7%) 10 Phage display (competitive panning) 1 PMID:26246327 Plasma of peanut-tolerant subject 3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3164 DVHKLGN(37.4%) AQSLETS(12.9%) NDIPHLM(12.3%) ETALIAA(7.5%) QQLAVDT(3.8%) RIHVHNS(3.5%) LYADSVL(3.2%) SVASTTV(3.2%) DLNMPIS(2.1%) IAGPFKL(1.6%) 10 Phage display (competitive panning) 2 PMID:26246327 Plasma of peanut-tolerant subject 3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3165 AQSLETS(40.5%) ETALIAA(16.6%) QQLAVDT(11.0%) DVHKLGN(10.8%) LYADSVL(7.7%) RIHVHNS(3.1%) NDIPHLM(2.8%) NPMYHSS(1.9%) KQEAMTL(1.1%) QNSPFSE(1.0%) 10 Phage display (competitive panning) 3 PMID:26246327 Plasma of peanut-tolerant subject 3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Bound phages were eluted by adding 25 μg Ara h 1 protein. The ten most frequent peptides were shown. 3166 RIRSEEL(24) TVVVAAG(17) VQLSIIV(15) PRTTIMG(15) KFVVAFC(14) LAPREGA(13) LMTARWC(13) FWALATW(13) YCVGDGC(13) WVNAATC(12) SRGVGVG(12) RRRPGAV(12) QLVRGRW(12) SVFIMLR(11) VFMGGCR(11) TSEGALR(11) SSRSSGG(11) GRLSADG(11) RRAVRFM(11) GLCAEAC(10) 20 0 PMID:22870226 Not determined. Not determined. Not determined. Not determined. pVIII fth1-dp-based X7 phage display library (X7) For the analysis of the naive libraries, four samples of 4.0e8 phages each were amplified individually by the PCR reactions. The amplified PCR products were validated for size (152 bp) by running in 2% agarose gel. All 4 PCR cleaned products were united and their concentration was measured. The sample was dried by speed vac and sent for Illumina sequencing. The 20 top most frequent peptides for each capture are given. 3167 ADGIVGW(3105) LAAGAVW(3039) SAGFAME(2944) VSLCSSR(2925) SVTDYVE(2855) RMGIRAL(2659) DDDGLDG(1931) SAARVFM(1723) QLYGARE(1716) NRSREMG(1707) IVPACRG(1646) RLVYVPS(1335) TGCSSIL(1166) AFHSGLT(1106) CSWTLER(996) GGGVGLL(912) WHAQVGF(852) ERRMGSC(844) SGVGATH(770) RLRWWGR(738) 20 Phage display (common panning) 1 PMID:22870226 Anti-gp120 monoclonal antibody GV4H3 Surface protein gp120, SU Not determined. Not determined. pVIII fth1-dp-based X7 phage display library (X7) the captured phages were eluted and directly amplified by PCR. The 20 top most frequent peptides for each capture are given. 3168 ADGIVGW(11489) SAGFAME(5173) LAAGAVW(3140) VTPHTGF(1172) GAHVAGG(672) LTACTGF(660) LGWAVLD(506) GTASVGF(444) SLGWAVP(355) GVGWAVP(344) AVGWAVP(336) GPGMALE(324) PASLVGF(300) SAGWALP(293) EVGWAVH(272) LNSMVGF(226) VGWAVLE(222) QMPNLGF(215) LVGWALS(202) SAGWALE(201) 20 Phage display (common panning) 2 PMID:22870226 Anti-gp120 monoclonal antibody GV4H3 Surface protein gp120, SU Not determined. Not determined. pVIII fth1-dp-based X7 phage display library (X7) the captured phages were eluted and directly amplified by PCR. The 20 top most frequent peptides for each capture are given. 3169 ADGIVGW(65003) SAGFAME(6630) VGWAVLE(2409) VTPHTGF(2366) LAAGAVW(2364) SAGVAME(2242) ADGIVGG(1009) EVGWAVH(905) WVGWAVQ(834) LGWAVLD(768) AVGWAVP(536) AGWAVLE(469) LVGWALS(453) SLGWAVP(445) GAHVAGG(413) LTACTGF(368) GVGFALD(362) GVGFALE(349) RVSAEVW(346) LAGWALD(337) 20 Phage display (common panning) 3 PMID:22870226 Anti-gp120 monoclonal antibody GV4H3 Surface protein gp120, SU Not determined. Not determined. pVIII fth1-dp-based X7 phage display library (X7) the captured phages were eluted and directly amplified by PCR. The 20 top most frequent peptides for each capture are given. 3170 ASSSHRS(73,075) DLKIPLR(37,802) YRAPWPP(34,250) AMSSRSL(15,301) LPLTPLP(13,006) IEFSPLM(4,765) GETRAPL(1,279) RHEPPLA(6) 8 Phage display (common panning) 3/4 PMID:22178910 Progenitor stem cell line KS483 at 4 days of differentiation Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) KS483 cells were incubated for 10 min on ice with 3 ml of 0.1 M HCl (pH 2.2) to elute cell surface-bound phage. 3171 GETRAPL(397,669) AMSSRSL(54,629) NPWTTRP(50,155) LPLTPLP(29,175) YRAPWPP(23,765) ASSSHRS(5,033) ALPQIVR(210) 7 Phage display (common panning) 3 PMID:22178910 Progenitor stem cell line KS483 at 4 days of differentiation Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) KS483 cells were lysed for 1 h on ice in 3 ml of 30 mM Tris–HCl and 1 mM EDTA (pH 8.0) to recover the cell-associated phage fraction. 3172 GETRAPL(3,599,322) ASSSHRS(57,476) YRAPWPP(56,436) DLKIPLR(55,342) IEFSPLM(43,697) LPLTPLP(20,988) AMSSRSL(11,517) 7 Phage display (common panning) 3 PMID:22178910 Progenitor stem cell line KS483 at 8 days of differentiation Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) KS483 cells were incubated for 10 min on ice with 3 ml of 0.1 M HCl (pH 2.2) to elute cell surface-bound phage. 3173 RHEPPLA(543,337) GETRAPL(427,153) AMSSRSL(134,645) YRAPWPP(55,732) ALPQIVR(34,034) ASSSHRS(24,902) WQSVPTI(20,390) LPLTPLP(20,003) DERHQHY(19,404) DLKIPLR(59) 10 Phage display (common panning) 1-4 PMID:22178910 Progenitor stem cell line KS483 at 8 days of differentiation Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) KS483 cells were lysed for 1 h on ice in 3 ml of 30 mM Tris–HCl and 1 mM EDTA (pH 8.0) to recover the cell-associated phage fraction. 3174 HAIYPRH(36) GKPMPPM(35) NHWASPR(33) SPPPPPP(32) TPPPPPP(31) VGYRLME(25) LPPPPPP(24) ATSAIHG(24) SPPVLPH(24) STYRITT(23) LSPLTPP(23) 699725 0 PMID:22178910 Not determined. Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Before analyzing the phage display libraries selected against biological targets, we screened for potential sequence biases introduced by the propagation of the phages in bacteria by sequencing the naive (unselected) library after one round of bacterial amplification. 3175 WPTDHQMLRIPM(342) DWSSWVYRDPQT(143) QSDPWLVSRWFA(66) TYPSTQWFFAKF(60) TLPPEAWWRTMY(52) YPSSEQLLAWWG(40) ELPSVEQLWDFF(37) WPWDPLRISDWL(36) TWVSDLDMWLGA(28) RLPTSMELLAAF(24) QDPWKVKNWINQ(15) FFPSEQVLIAAL(12) KLPSPYDLYLFL(9) WDPWQIDRWVLL(8) NAPSIYDWLATL(5) WPLWSFDWPQNA(4) WLTEDMSAFMSY(4) DPWFINEWIRIP(4) WPSDNQLLQIHV(2) SWDPLLVSAWIK(1) 20 0 PMID:26161996 Not determined. Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The Ph.D. library was amplified once with no selection and used as the unselected library control (replicate 1). 3176 WPTDHQMLRIPM(342) QSDPWLVSRWFA(66) TYPSTQWFFAKF(60) TLPPEAWWRTMY(52) SFPDVQWWRNQY(50) YPSSEQLLAWWG(40) ELPSVEQLWDFF(37) WPWDPLRISDWL(36) TWVSDLDMWLGA(28) RLPTSMELLAAF(24) SDSWVRGLNYWY(16) SYWVPDIVWAGL(15) QDPWKVKNWINQ(15) GLPSSAELERLW(14) LPWPSDQIILMW(14) FFPSEQVLIAAL(12) LPSSAELLWALR(9) WDPWQIDRWVLL(8) FDPLNIRHWIWG(7) DPWFINEWIRIP(4) 20 0 PMID:26161996 Not determined. Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The Ph.D. library was amplified once with no selection and used as the unselected library control (replicate 2). 3177 DWSSWVYRDPQT(3030) WPLWSFDWPQNA(2) 2 0 PMID:26161996 Not determined. Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The Ph.D. library was amplified four times with no selection and used as the unselected library control (replicate 1). 3178 WPTDHQMLRIPM(40.15%) QSDPWLVSRWFA(8.55%) DWSSWVYRDPQT(7.82%) TYPSTQWFFAKF(1.90%) ELPSVEQLWDFF(1.87%) YPSSEQLLAWWG(1.74%) WPSDNQLLQIHV(1.64%) RLPTSMELLAAF(1.64%) TWVSDLDMWLGA(1.53%) WDPWQIDRWVLL(1.47%) NAPSIYDWLATL(1.20%) KLPSPYDLYLFL(1.15%) TLPPEAWWRTMY(0.72%) WPWDPLRISDWL(0.62%) QDPWKVKNWINQ(0.56%) FFPSEQVLIAAL(0.43%) SWDPLLVSAWIK(0.43%) WPLWSFDWPQNA(0.38%) WLTEDMSAFMSY(0.34%) DPWFINEWIRIP(0.32%) 20 Phage display (subtractive panning) 4 PMID:26161996 IL-4-polarized M2 macrophage Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Subtractive phage display using murine IL-4- polarized M2 ("anti-inflammatory") macrophage for positive selection and IFN-γ- and LPS-polarized M1 ("pro-inflammatory") macrophage for negative selection was completed in biological duplicates with one initial enrichment selection followed by three rounds of paired negative/positive selection. Barcoded phage library amplicons were sequenced by Illumina, and the top 20 most abundant sequences from the last rounds are shown （replicate 1). 3179 WPTDHQMLRIPM(32.14%) TYPSTQWFFAKF(9.51%) SFPDVQWWRNQY(5.16%) WPWDPLRISDWL(5.08%) TLPPEAWWRTMY(4.42%) QSDPWLVSRWFA(3.59%) YPSSEQLLAWWG(3.52%) RLPTSMELLAAF(3.21%) GLPSSAELERLW(2.47%) ELPSVEQLWDFF(2.44%) TWVSDLDMWLGA(1.89%) QDPWKVKNWINQ(1.78%) FFPSEQVLIAAL(1.64%) SDSWVRGLNYWY(1.27%) FDPLNIRHWIWG(1.20%) WDPWQIDRWVLL(1.19%) LPWPSDQIILMW(1.17%) DPWFINEWIRIP(0.88%) LPSSAELLWALR(0.81%) SYWVPDIVWAGL(0.74%) 20 Phage display (subtractive panning) 4 PMID:26161996 IL-4-polarized M2 macrophage Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Subtractive phage display using murine IL-4- polarized M2 ("anti-inflammatory") macrophage for positive selection and IFN-γ- and LPS-polarized M1 ("pro-inflammatory") macrophage for negative selection was completed in biological duplicates with one initial enrichment selection followed by three rounds of paired negative/positive selection. Barcoded phage library amplicons were sequenced by Illumina, and the top 20 most abundant sequences from the last rounds are shown (replicate 2). 3180 SCNVAGGTCGLT YPSSEQLLAWWG VAGPQITKYNVP ESSPATKFVRPS VCLYVNIECSQQ LGPQRQENSHPF GLPIWPQHLTNT RMDPDQPRARVA GFGGLVDDVRGG INIPDEQSSNHL QSDPWLVSRWFA ALVNQADGDSTQ SLYGKKSPMGYP YSDEGTPRSGGL WPTDHQMLRIPM 15 Phage display (subtractive panning) 4 PMID:26161996 IL-4-polarized M2 macrophage Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Subtractive phage display using murine IL-4- polarized M2 ("anti-inflammatory") macrophage for positive selection and IFN-γ- and LPS-polarized M1 ("pro-inflammatory") macrophage for negative selection was completed in biological duplicates with one initial enrichment selection followed by three rounds of paired negative/positive selection. From the last round of each duplicate phage display experiment, 20 phage plaques were randomly selected for Sanger sequencing (replicate 1). 3181 WPTDHQMLRIPM ELPSVEQLWDFF WPTDHQMLRIPM TLPPEAWWRTMY MLPSPELLWEWL WDPWQIDRWVLL GLPSSAELERLW TYPSTQWFFAKF FFPSEQVLIAAL GLPSSAELERLW QDPWKVKNWINQ WPTDHQMLRIPM GLPSSAELERLW WPWDPLRISDWL WDPWQIDRWVLL TYPSTQWFFAKF ELPSVEQLWDFF YPSDLDILRLFS GLPSSAELERLW SDSWVRGLNYWY 20 Phage display (subtractive panning) 4 PMID:26161996 IL-4-polarized M2 macrophage Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Subtractive phage display using murine IL-4- polarized M2 ("anti-inflammatory") macrophage for positive selection and IFN-γ- and LPS-polarized M1 ("pro-inflammatory") macrophage for negative selection was completed in biological duplicates with one initial enrichment selection followed by three rounds of paired negative/positive selection. From the last round of each duplicate phage display experiment, 20 phage plaques were randomly selected for Sanger sequencing (replicate 2). 3182 CPLHARLPC(82.49%)[2.16693±0.28562] CFDTRSLVC(5.05%)[1.46493±0.08252] CDHGYLPSC(0.92%)[0.72612±0.07109] CHYDGARAC(0.81%)[1.04729±0.07235] 4 Phage display (subtractive panning) 5 PMID:24265677 Mycobacterium tuberculosis △leucineD and △panthothenateCD double auxotroph, △leu/△pan Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Microtiter plate wells were coated with 100 ml of mycobacteria suspension, with an optical density of 1.0 at 660 nm in carbonate buffer, and incubated overnight at 4uC. Absorbance was determined using a microtiter plate reader at the wavelength of 405 nm. Five rounds of biopanning were performed. The first three rounds were targeted against M. tb (△leu/△pan), followed by a subtraction round against M. smegmatis. The fifth round of positive selection was against the targeted M. tb (△leu/△pan). We further showed that the clone displaying the CPLHARLPC peptide which was identified by Illumina sequencing as the most enriched, binds better to mycobacteria than three clones selected by random picking. Using surface plasmon resonance, we showed that chemically synthesised CPLHARLPC peptide binds to a 15 KDa peptide from M.tb H37Rv whole cell lysates. 3183 QCVPLKCLWDRCE(110) TCLCKRCIKELCC(36) VCERQVCYLMSCW(22) YCVWDKCLWLMCE(16) ACDARPCPQTYCL(12) ACGMSICVLYGCN(9) 6 Phage display (common panning) 2 PMID:25348396 Human beta-FXIIa Not determined. Not determined. Not determined. XCX4CX4CX phage display library Identical peptides would be considered as target-specific peptides. We repeated a first round of selection against FXIIa and found that only six peptide sequences were common in both pools. Four of them, QCVPLKCLWDRCE, ACDARPCPQTYCL, TCLCKRCIKELCC and YCVWDKCLWLMCE, corresponded to confirmed binding motifs that were previously found after three rounds of selection. 3184 ACKRTHCLPPCCGGSG(743) ACPLLPPCADDCGGSG(701) ACSRHCLTLPPCGGSG(364) ACPLLPPCSLDCGGSG(317) ACRQLPPCSFECGGSG(302) ACSWRCPSLPPCGGSG(194) ACSVLPPCSFSCGGSG(148) ACNTLCPYLPPCGGSG(146) ACYQLPPCDHSCGGSG(140) ACPILPPCHAHCGGSG(135) ACHLPPCSLHLCGGSG(125) LPP 6243 Phage display (common panning) 1 PMID:25348396 Sortase A from S. aureus (SrtA) Not determined. Not determined. Not determined. CX3CX4CG, CX4CX3CG, CX4CX4CG, CX3CX5CG and CX5CX3CG phage display library pool 3185 MAACSILPPCNPPQCGGSG(5860) MAACPLLPPCHLPQCGGSG(4327) MAACTLLPPCTPDQCGGSG(3990) MAACPELPPCQLMLCGGSG(2326) MAACQVLPPCGLQLCGGSG(2302) MAACRPKQCWQLPPCGGSG(1798) MAACSFIQDCHPQSCGGSG(1574) MAACPMLPPCDLSYCGGSG(1258) MAACRQLPPCAEYVCGGSG(1231) MAACLPPHSCWNQVCGGSG(1227) MAACPSLPPCWQLQCGGSG(1131) LPP 2776 Phage display (common panning) 1 PMID:25348396 Sortase A from S. aureus (SrtA) Not determined. Not determined. Not determined. CX3CX6CG, CX6CX3CG, CX4CX5CG and CX5CX4CG phage display library pool 3186 MAACTQSACSARVVCGGSG(5410) MAACAPDQCTKFTMCGGSG(2493) MAACTYALCTARTFCGGSG(1905) MAACSASQCSARIGCGGSG(1493) MAACMLSGSCTARSCGGSG(1381) MAACKHSDCTARFPCGGSG(1315) MAACFRYQCTARSHCGGSG(1081) MAACPVKPSCHSGRCGGSG(1056) MAACPLSACSGRTLCGGSG(1026) MAACAYRSCQGTARCGGSG(918) MAACWTPTCSARSHCGGSG(890) [TS]-A-R, [KR]-[FY]-[ST]-L 6906 Phage display (common panning) 1 PMID:25348396 Urokinase-type plasminogen activator Urokinase plasminogen activator surface receptor Not determined. Not determined. CX3CX6CG, CX6CX3CG, CX4CX5CG and CX5CX4CG phage display library pool 3187 HCWVSACRHWRCQSHS(795) ACESLTCALIVCQSHS(434) VCLRQTCSSANCGSHS(425) VCDQWACGQEWCVSHS(381) RCTGRSCEWHACGSHS(369) QCWQMPCSLGSCPSHS(344) WCWYIGCAGIGCASHS(322) KCSDQPCELMACKSHS(319) ACDARPCPQTYCLSHS(307) SCISRKCLAMQCHSHS(300) GCGAVACCLRSCQSHS(300) RPCP, VXXKCL 17469 Phage display (common panning) 1 PMID:25348396 Human beta-FXIIa Not determined. Not determined. Not determined. XCX4CX4CX phage display library 3188 MADCWDSVCYRVLCWSHS(704) MADCYQLCRVSCESHS(501) MAMCEQRACTFRECWSHS(417) MARCFIYCRTLCMSHS(415) MANCFQTCRVSCYSHS(350) MAGCFTPCRTMCWSHS(317) MAWCATDLCDACVCQSHS(309) MATCSWGKCQVTDCGSHS(295) MAQCWIACRVVCLSHS(292) MAWCHTSCQVACSSHS(290) MARCLGHRCGAWICSSHS(287) [FYW]-X-X-C-R-V 3532 Phage display (common panning) 1 PMID:25348396 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX3CX3CX and XCX4CX4CX phage display library pool 3189 MAECISSVCTRGVCLSHS(1791) MATLSGALH*AVYFSL(1248) MAKCWRGDCALSVCASHS(1064) MASCCACVGFVCLTP(808) MASCEACLCMGLCVLLTP(804) MAHCAYLRCTLLLCQSHS(798) MASCRDVCCSHCLSHS(795) MAGCPEAACCLQVCGSHS(754) MAYCTLGCDTDCSSHS(718) MATCRQGCVGACSSHS(718) MAQCQTRCTAVCASHS(702) HPQ 498 Phage display (common panning) 1 PMID:25348396 Streptavidin Biotin Not determined. Not determined. XCX3CX3CX and XCX4CX4CX phage display library pool 3190 RVRSAPSSS(17)[2.014 ± 0.067] RVRSYSPHL(6)[NT] RVRSNAASM(3)[NT] RVRSSPVVQ(1)[NT] RVRSIPHVA(1)[NT] R-V-R-S-P-S 5 Phage display (subtractive panning) 4 PMID:28654884 Staphylococcus aureus, S.aureus Not determined. Not determined. Not determined. pVIII-9aa phage display library (X9) ELISA Phage particles bearing no recombinant insert, from superinfection of phagemid vector pC89-containing cells, served as a negative control for evaluation of background from non-specific binding.The ability of the isolated phage clone to interact specifically with S. aureus and the efficacy of its bacteria-binding properties were established by ELISA on a kinetic plate reader at 405 nm. The library was used for pretreatment with the plastic materials which will be used to the selection protocolon-binding phage clones. A phage clone displaying peptide capable of specific binding to S. aureus cell surface, namely St.au9IVS5 (sequence peptide RVRSAPSSS) was selected from a 9-mer phage peptide library. The ability of the isolated phage clone to interact specifically with S. aureus and the efficacy of its bacteria-binding properties were established by using enzyme linked immune-sorbent assay (ELISA). We also demonstrated by Western blot analysis that the most reactive and selective phage peptide binds a 78 KDa protein on the bacterial cell surface. Furthermore, we observed selectivity of phage–bacteria-binding allowing to identify clinical isolates of S. aureus in comparison with a panel of other bacterial species. In order to explore the possibility of realizing a selective bacteria biosensor device, based on immobilization of affinity-selected phage, we have studied the physisorbed phage deposition onto a mica surface. Atomic Force Microscopy (AFM) was used to determine the organization of phage on mica surface and then the binding performance of mica-physisorbed phage to bacterial target was evaluated during the time by fluorescent microscopy. The system is able to bind specifically about 50% of S. aureus cells after 15' and 90% after one hour. Due to specificity and rapidness, this biosensing strategy paves the way to the further development of new cheap biosensors to be used in developing countries, as lab-on-chip (LOC) to detect bacterial agents in clinical diagnostics applications. 3191 LSSTTSPRVTTS(6)[0.889 ± 0.016] QQLPSSSTSTYP(5)[1.011 ± 0.031] TVAHHYPPQTPL(5)[0.832 ± 0.016] ITTGNEWSYSRK(4)[0.846 ± 0.054] NKLPMPISVPLV(4)[0.995 ± 0.016] TELGQVSNAIPW(3)[0.932 ± 0.021] 6 Phage display (subtractive panning) 4 PMID:28634746 SiHa cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The reaction was terminated by adding 2 M H2SO4 (50 ll/well) and measured at 450 nm by an ELISA reader.Irrelevant phage clone (IRP, an amplified phage clone randomly selected from the original phage-displayed peptide library) and PBS were used as negative controls. Data shown (OD450) were reproduced from Figure 1 in the reference. SiHa and HEK293 cells were used as positive target cells and negative absorber cells. Phages were incubated with HEK293 cells for 1 h after which the supernatant containing unbound phages was incubated with the blocked SiHa cells. After sequencing of positive clones, six different peptide sequences were obtained and CSP3 (QQLPSSSTSTYP) showed best affinity and specificity to SiHa cells via immunofluorescence assay. Peptide CSP3 may be a potential candidate for molecular imaging detection and targeting therapy of cervical cancer. 3192 SHGRITFAYFAN[3.187 ± 0.128] KPSYNQLYQLRF[NT] HSTLHYRFSDWP[NT] WDTKAVTKVPSW[NT] NIHAHETPFKMV[NT] AEYLANPESSEA[NT] HVVSKVLTDRTD[NT] YMPYMPHLPQEH[NT] WADKEHHLILRN[NT] SDNLLIKARTSY[NT] TPQSIGKAFQSE[NT] LSTPNTVGLARF[NT] FISVQNSFGIEI[NT] KVATIRDLHSMT[NT] TEYKKGHDPAEI[NT] VTGLTPYISESG[NT] HPIQSLTTNQTK[NT] DNDENITWGLTR[NT] AFNAHGALLFVT[NT] KVWDFGYAFGRT[NT] TERMSLTTRIGF[NT] KVWSIPYAFMAR[NT] VKVHSGQEHETW[NT] 23 Phage display (common panning) 4 PMID:28635259 Hemagglutinin, HA Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Triplicate determinations were performed at each data point, and the average OD450 nm values were determined. Data shown were reproduced from Figure 1 in the reference. The final selected haemagglutinin-binding peptide, SHGRITFAYFAN, was conjugated to an anti-influenza A virus (IAV) antisense oligonucleotide (ODN). The delivery efficiency and the anti-IAV effects of the conjugated molecule were evaluated in a cell culture and a mouse infection model. The conjugated molecule was successfully delivered into IAV-infected host cells more efficiently than the anti-IAV ODN in vitro and in vivo. Furthermore, the conjugated molecule protected 80% of the mice from lethal challenge and inhibited the plaque count by 75%, compared to the un-conjugated molecule (60% and 40%). These findings demonstrate that the delivery of antisense oligodeoxynucleotides to infected tissues by a virus-binding peptide-mediated system is a potential therapeutic strategy against IAV. 3193 LYAKPRDLSSLT HDGVSANSSIHA STWLYPLTSLQL IAKQTVTSSSHP HYSDKWGHAGTE NNPDPHYKSTWP KAHYGFMHQLSS MNGSPDSSQQKS DADYTGYLDLND GYTLQYTLPGAP FHESWPTFLSPS YCGGDSCLVYVD DRTAFYSPYNLS FHWPWWDTYVGK MPSSSGMMIRTT CVEFFAAVSESS 16 Phage display (common panning) 4 PMID:28635259 Neuraminidase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Triplicate determinations were performed at each data point, and the average OD450 nm values were determined. Data shown were reproduced from Figure 1 in the reference. 3194 SRTGNWTRIDQS(3) ARYFARHPTAMG(1) DGQSDLSPRPPH(1) FATSRLIDTLAS(1) GSSSKGSAFVTA(1) HTRQWSGSGQMF(1) INPYAQHPTGSI(1) KASFYNVTSLNE(1) NSVHRAALGPGT(1) NTLLLNMDPSVV(1) RLIDKKPDAFIT(1) SRTGIWTRTDRS(1) WDPWSRVMGPNT(1) WNGPKLVSNGDG(1) YAGEPVSVPGTS(1) YRAGQVVMNVGA(1) 16 Phage display (common panning) 3 PMID:28482148 Eu2O3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The high efficiency and the specificity of binding Eu2O3 nanoparticles by M13 PG-Eu-3 was further confirmed with the use of atomic force microscopic (AFM) imaging. It is easy to see that M13 PG-Eu-3 phages bind to Eu2O3 NPs with one end of the virion, where the SRTGNWTRIDQS peptide is presented. The peptide revealed the ability to catalyze Eu2O3 nanoparticles’ formation from Eu(OH)3 and Eu(NO3)3 solutions. 3195 LGRFYAASG 1 Phage display (common panning) 5 PMID:28652618 Annexin A2 Not determined. Not determined. Not determined. X4YX4 phage display library Annexin A2 and a cell-internalizing, penetratin-fused version of the selected peptide (LGRFYAASG-pen) co-localized and specifically accumulated in the cytoplasm at the cell edges and cell-cell contacts. Functionally, tumor cells incubated with LGRFYAASG-pen showed disruption of filamentous actin, focal adhesions and caveolaemediated membrane trafficking, resulting in impaired cell adhesion and migration in vitro. These effects were paralleled by a decrease in the phosphorylation of both focal adhesion kinase (Fak) and protein kinase B (Akt). Likewise, tumor cells pretreated with LGRFYAASG-pen exhibited an impaired capacity to colonize the lungs in vivo in several mouse models. 3196 CNHQHQKGC(1)[108, 37.23] CQNAHQKGC(1)[2.4, 4.18] CQSHKNNKC(1)[N.A., N.A.] CHDKQSKKC(1)[N.A. N.A.] 4 Phage display (common panning) 4 PMID:28650615 Human Hair Not determined. Not determined. Not determined. ACX7CG phage display library ELISA Phage enzyme-linked immunosorbent assay (ELISA) experiments were performed in order to determine the apparent binding strength (Kapp, 1.0e10 M-1) and relative affinity (RAmax) of selected phage clones in comparison to the random phage library. The first column of the affinity value is Kapp, and the second RAmax. The hair samples were incubated with the shampoo formulations containing the phage library. Washing was performed using a surfactant concentration of 0.1 w/w% in each round. A peptide with the sequence of CNHQHQKGC that can bind to human hair under shampooing conditions was used to enhance the deposition of model fragrance delivery systems. These delivery systems are either based on poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA) copolymers as a representative for polymeric profragrances or polyurethane/urea-type core-shell microcapsules as a model physical fragrance carrier. The incorporation of a hair binding peptide enhanced the deposition of PHPMA copolymers with a factor of 3.5-5.0 depending on the extent of peptide incorporation, whereas 10 w/w% surface functionalization of microcapsules with the peptide led to a 20-fold increase in their deposition. In a final experiment, treatment of the hair samples under realistic application conditions with the peptide-functionalized microcapsules resulted in an increase fragrance release from the hair surfaces. 3197 CNHQHQKGC(3)[108, 37.23] CKSKNHPSC(1)[21.0, 18.44] CQNAHQKGC(1)[2.4, 4.18] CGHNKNKDC(1)[N.A., N.A.] 4 Phage display (common panning) 5 PMID:28650615 Human Hair Not determined. Not determined. Not determined. ACX7CG phage display library ELISA Phage enzyme-linked immunosorbent assay (ELISA) experiments were performed in order to determine the apparent binding strength (Kapp, 1.0e10 M-1) and relative affinity (RAmax) of selected phage clones in comparison to the random phage library. The first column of the affinity value is Kapp, and the second RAmax. The hair samples were incubated with the shampoo formulations containing the phage library. Washing was performed using a surfactant concentration of 0.1 w/w% in each round. A peptide with the sequence of CNHQHQKGC that can bind to human hair under shampooing conditions was used to enhance the deposition of model fragrance delivery systems. These delivery systems are either based on poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA) copolymers as a representative for polymeric profragrances or polyurethane/urea-type core-shell microcapsules as a model physical fragrance carrier. The incorporation of a hair binding peptide enhanced the deposition of PHPMA copolymers with a factor of 3.5-5.0 depending on the extent of peptide incorporation, whereas 10 w/w% surface functionalization of microcapsules with the peptide led to a 20-fold increase in their deposition. In a final experiment, treatment of the hair samples under realistic application conditions with the peptide-functionalized microcapsules resulted in an increase fragrance release from the hair surfaces. 3198 CNHQHQKGC(3)[108, 37.23] CKSKNHPSC(2)[21.0, 18.44] CQSHKNNKC(1)[N.A., N.A.] 3 Phage display (common panning) 4 PMID:28650615 Human Hair Not determined. Not determined. Not determined. ACX7CG phage display library ELISA Phage enzyme-linked immunosorbent assay (ELISA) experiments were performed in order to determine the apparent binding strength (Kapp, 1.0e10 M-1) and relative affinity (RAmax) of selected phage clones in comparison to the random phage library. The first column of the affinity value is Kapp, and the second RAmax. The hair samples were incubated with the shampoo formulations containing the phage library. The washing stringency was gradually enhanced by stepwise increasing the surfactant concentration from 0.1 w/w% (Round 1) to 0.4 w/w% (Round 4). A peptide with the sequence of CNHQHQKGC that can bind to human hair under shampooing conditions was used to enhance the deposition of model fragrance delivery systems. These delivery systems are either based on poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA) copolymers as a representative for polymeric profragrances or polyurethane/urea-type core-shell microcapsules as a model physical fragrance carrier. The incorporation of a hair binding peptide enhanced the deposition of PHPMA copolymers with a factor of 3.5-5.0 depending on the extent of peptide incorporation, whereas 10 w/w% surface functionalization of microcapsules with the peptide led to a 20-fold increase in their deposition. In a final experiment, treatment of the hair samples under realistic application conditions with the peptide- functionalized microcapsules resulted in an increase fragrance release from the hair surfaces. 3199 CNHQHQKGC(3)[108, 37.23] CKSKNHPSC(2)[21.0, 18.44] CQNAHQKGC(2)[2.4, 4.18] CGHNKNKDC(1)[N.A., N.A.] CHDKQSKKC(1)[N.A., N.A.] 5 Phage display (common panning) 5 PMID:28650615 Human Hair Not determined. Not determined. Not determined. ACX7CG phage display library ELISA Phage enzyme-linked immunosorbent assay (ELISA) experiments were performed in order to determine the apparent binding strength (Kapp, 1.0e10 M-1) and relative affinity (RAmax) of selected phage clones in comparison to the random phage library. The first column of the affinity value is Kapp, and the second RAmax. The hair samples were incubated with the shampoo formulations containing the phage library. The washing stringency was gradually enhanced by stepwise increasing the surfactant concentration from 0.1 w/w% (Round 1) to 0.5 w/w% (Round 5). A peptide with the sequence of CNHQHQKGC that can bind to human hair under shampooing conditions was used to enhance the deposition of model fragrance delivery systems. These delivery systems are either based on poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA) copolymers as a representative for polymeric profragrances or polyurethane/urea-type core-shell microcapsules as a model physical fragrance carrier. The incorporation of a hair binding peptide enhanced the deposition of PHPMA copolymers with a factor of 3.5-5.0 depending on the extent of peptide incorporation, whereas 10 w/w% surface functionalization of microcapsules with the peptide led to a 20-fold increase in their deposition. In a final experiment, treatment of the hair samples under realistic application conditions with the peptide- functionalized microcapsules resulted in an increase fragrance release from the hair surfaces. 3200 YPLRPNAESLRF DPIYALSWSGMA NATHLTADHVNK YPSAPPQWLTNT AQHIRSWDGFSH LPLYTSPDKPGK SILNYPTNPGIA LLADTTHHRPWT HPIRVQPDWGFL VETHTTQWLITEV TPMGRPHPETPA SGKTSLISHAAL NHLPLPPPAATM SDTQMPPAXGRA GYLPLHSITYRP QLLEPVNLSTGP WHPPKGLSPLPD FRLPGSLINHPQ SILSTMSPHGAT NYNPHNPFPPAP NHT, [ST]-[IV]-L-S 20 Phage display (subtractive panning) 3 PMID:28453953 Human bone marrow stromal cell (hBMSC) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The NEB 12-mer peptide library was prescreened against sintered Hydroxyapatite (HAP) disks prior to introduction to the cells. A peptide sequence with high affinity towards apatite (VTKHLNQISQSY, VTK) was identified using phage display. In this study, a combinatorial phage display identified the cell binding sequence (DPIYALSWSGMA, DPI) which was combined with the mineral binding sequence to generate the dual peptide DPI-VTK. DPI-VTK demonstrated significantly greater binding affinity (1/KD) to apatite surfaces compared to VTK, phosphorylated VTK (VTKphos), DPIVTKphos, RGD-VTK, and peptide-free apatite surfaces (p < 0.01), while significantly increasing hBMSC adhesion strength (t50, p < 0.01). MSCs demonstrated significantly greater adhesion strength to DPI-VTK compared to other cell types, while attachment of MC3T3 pre-osteoblasts and murine fibroblasts was limited (p < 0.01). MSCs on DPI-VTK coated surfaces also demonstrated increased spreading compared to pre-osteoblasts and fibroblasts. MSCs cultured on DPI-VTK coated apatite films exhibited significantly greater proliferation compared to controls (p < 0.001). Moreover, early and late stage osteogenic differentiation markers were elevated on DPI-VTK coated apatite films compared to controls. 3201 FPWYKWRLPDVS(3)[30.98 ± 5.84] IPWHRPAQVMHL(3)[27.31 ± 16.09] FPHHKQRYVDPL(3)[6.15 ± 3.98] MHKPTVHIKGPT(2)[21.93 ± 1.84] HTSSLWHLFRST(1)[4.92 ± 0] GMKPWFYSNWKG(1)[1.70 ± 0] GMLHWSYSIFNP(1)[2.01 ± 0] FHRAPWJYLGNY(1)[10.89 ± 8.28] IPLHSLHVKWGK(1)[2.01 ± 0] MGKSTLRYTTIV(1)[31.28 ± 15.03] 10 Phage display (common panning) 3-5 PMID:27861731 Enargite Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Competition experiment Binding assays were performed with selected enargite phage clones in competition with the Wt phage. In these experiments, equal amounts of each phage were mixed and then the usual binding assay was performed. In another experiment, an increasing amount of wild-type phage was added to a constant amount of enargite-binding phage and the assay was performed as with the mini library screen. Screening of a random peptide phage display library resulted in the identification of an enargite-selective peptide with the sequence MHKPTVHIKGPT. Mineral-binding selectivity was demonstrated by binding studies, zeta potential determination and immunochemistry. Peptides that have the ability to discriminate between enargite and chalcopyrite provide a greener option for the separation of arsenic containing contaminants from copper concentrates. This represents the first step towards a major advance in the replacement or reduction of toxic collectors as well as reducing the level of arsenic-bearing minerals in the early stages of mineral processing. 3202 DKKKCDGKRCSWPS(3)[23.91 ± 0] RKKKCKGNCCYTPQ(3)[220.18 ± 55.53] ERSGCHKKACPKTP(2)[68.64 ± 0] HKTQCNPRACTRRH(1)[1.03 ± 0] KKTNCKHDSCRTCQ(1)[12.98 ± 0] QRNKCHHNTCVKML(1)[187.40 ± 87.28] EKDRCTKNTCKPPA(1)[219.02 ± 149.36] KDHDCHRAQCRPNL(1)[41.39 ± 0] GKKKCPNKSCTSLF(1)[10.80 ± 0] KSKSCEAMQCNKYY(1)[5.40 ± 0] KNKRCSQGCCINNG(1)[5.40 ± 0] KNEKCAHHKCYLYP(1)[10.80 ± 0] GKCSCKEKQCRTTL(1)[7.58 ± 0] KSRHCSQIQCGDKV(1)[4.24 ± 0] EKKKCGTMACPYRA(1)[5.19 ± 0] 15 Phage display (common panning) 3-5 PMID:27861731 Chalcopyrite Not determined. Not determined. Not determined. F88-FUSE X4CX4CX4 phage display library (X4CX4CX4) Competition experiment Binding assays were performed with selected chalcopyrite phage clones in competition with the Wt phage. In these experiments, equal amounts of each phage were mixed and then the usual binding assay was performed. In another experiment, an increasing amount of wild-type phage was added to a constant amount of chalcopyrite-binding phage and the assay was performed as with the mini library screen. Screening of a random peptide phage display library resulted in the identification of a chalcopyrite-selective peptide with the sequence RKKKCKGNCCYTPQ. Mineral-binding selectivity was demonstrated by binding studies, zeta potential determination and immunochemistry. Peptides that have the ability to discriminate between enargite and chalcopyrite provide a greener option for the separation of arsenic containing contaminants from copper concentrates. This represents the first step towards a major advance in the replacement or reduction of toxic collectors as well as reducing the level of arsenic-bearing minerals in the early stages of mineral processing. 3203 RCQYPLCS(14/40)[102.96 ± 23.15] RCLRSHCG(3/40)[4.60 ± 0] SCKTVFCY(3/40)[3.77 ± 0] RCPRFSCW(2/40)[10.51 ± 0.40] SCFRPTCP(2/40)[5.01 ± 0] RCQYSPCH(1/40)[4.70 ± 1.99] 6 Phage display (common panning) 3 PMID:27987304 LaPO4:Ce3+,Tb3+, LAP Not determined. Not determined. Not determined. LX-4 phage display library (XCX4CX) Binding assay The percent bound was calculated by dividing the number of eluted phage by the number of input phage and multiplying by 100. Results in Figure 1 are presented as the fold increase above the percent bound of the wild type bacteriophage from up to 6 independent experiments. Data shown were regenerated from Figure 1 in the reference. After three rounds of enrichment, a phage clone containing the surface peptide loop RCQYPLCS was found to bind specifically to LAP. Specificity and affinity of the identified phage bound peptide was confirmed by using binding and competition assays, immunofluorescence assays and zeta potential measurements. Binding and immunofluorescence assays identified the peptide’s affinity for the fluorescent phosphor components CAT (CeMgAl11O19:Tb3+) and BAM (BaMgAl10O17:Eu2+). No affinity was found for other fluorescent phosphor components such as YOX (Y2O3:Eu3+). The binding specificity of the RCQYPLCS peptide loop was improved 3-51-fold by using alanine scanning mutagenesis. The identification of peptides with high specificity and affinity for special components in the fluorescent phosphor in CFLs provides a potentially new strategic approach to rare earth recycling. 3204 GLHTSATNLYLH(7/10)[0.38 ± 0.28] AGYPLSENFYYP(1/10)[4.08 ± 0.31] WQDFGAVRSTRS(1/10)[3.15 ± 0.36] FHPRLQQDHWLH(1/10)[3.92 ± 0.44] 4 Phage display (competitive panning) 3 PMID:28094103 Particulate matter, PM Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay Spot intensity measurements (with background subtraction) using Image Quant. The data shown were reproduced from the Figure 1B in the reference. Bound phages were eluted using a PM suspension [100 μg PM/ml TBS (50 mM Tris-HCl (pH 7.5), 150 mM NaCl)]. Four PM binding peptides were obtained by phage display and binding characteristics of these peptides were investigated using the peptide array. The strongest binding peptide, WQDFGAVRSTRS, displayed a binding property, measured in terms of spot intensity, 11.4 times higher than that of the negative control, AAAAA. Inductively coupled plasma mass spectrometry (ICPMS) analysis of the transition metal compounds in the PM bound to the peptide spots was performed, and two peptides showed higher binding towards Cu and Zn compounds in PM. These results suggest that the screened peptides could serve as an indicator of transition metal compounds, which are related to adverse health effects, contained in PM. 3205 CHMQGRSTC CHGGQTVAC CNAGHLSQC CTTQMGYSC CPEWFRWHC CNSSASKNC CGWKHEQTC CVPSKPGLC CNSHRHGAC CWWNNFKHC CFPTGTYWC CALHSGQKC CTNASQSYC CNWMINKWC CRDLSLHSC CRTXNGTRC CFSAVVEPC CYKPVRVHC CSLFSKNYC 19 Phage display (common panning) 3 PMID:28414460 SECp43180 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 3206 DDGAYTHRSNLI GIWYRDVVQQWP SPGNQTYSSQVR GHGMLILSPNPT GSMAGPENHRAI EPAILTDAMPFN YAPDLLPNSTWT SHVDSASLRYWR SGVYKVAYDWQH GLHTSATNLYLH VHWDFRQWWQPS 11 Phage display (common panning) 3 PMID:28414460 SECp43180 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3207 TFQAFDLSPFPS(8/17)[2.11 ± 0.07] ISSPRSAPTPPY(6/17)[1.81 ± 0.01] HAPQTFSSPKFP(1/17)[0.34 ± 0.10] WPTYLNPSSLKA(1/17)[0.34 ± 0.07] KLWDIKLPQTSV(1/17)[0.27 ± 0.09] 5 Phage display (common panning) 4 PMID:28349164 Major structural protein VP28 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of the positive phage clones to the Envelope protein were measured by phage ELISA. The OD 450nm values were measured and data shown were re-generated from the Figure 1a in the reference. A bacteriophage clone VP28-4L was obtained, and its binding to purified rVP28 protein as well as WSSV from infected shrimp Litopaeneus vannamei tissue was confirmed by ELISA and western blot. The apparent equilibrium dissociation constant (Kd,app) was calculated to be 810 nM. VP28-4L did not show cross-reactivity with any other shrimp viruses. A 12-mer peptide (pep28, with the sequence ′TFQAFDLSPFPS′) displayed on the VP28-4L was synthesized, and its diagnostic potential was evaluated in a lateral flow assay (LFA). Visual detection of WSSV could be achieved using biotinylated-pep28 and streptavidin-conjugated gold nanoparticles. In LFA, 12.5 μg/mL of the virus could be detected from L. vannamei gill tissue homogenate within 20 min. Pep28 thus becomes an attractive candidate in bio-recognition of WSSV in field-usable diagnostic platforms benefitting the aquaculture sector. 3208 KPSHT(12) VNTSN(5) ASSHN(3) 3 Phage display (in vivo) 5 PMID:28359814 Tumor vessels Not determined. Not determined. Not determined. Ph.D.-5 M13 phage display library (X5) Biopanning of the phage-displayed peptide library was performed by in vitro biopanning using human endothelial progenitor cells (hEPCs) and in vivo biopanning in DAS model mice for a total of 3 and 2 rounds, respectively. Phage clones displaying ASSHN peptide showed a marked affinity for hEPCs in vitro, and also for tumor vessels in vivo. PEGylated liposomes modified with the ASSHN peptide (ASSHN-Lip) were designed and prepared for the delivery of anticancer agents. Confocal images showed that ASSHN-Lip clearly bound to hEPCs in vitro and tumor vessels, and also showed extravasation from the vessels. The administration of doxorubicin- encapsulated ASSHN-Lip into Colon26 NL-17-bearing mice significantly suppressed tumor growth compared with doxorubicin-encapsulated PEGylated liposomes. These results suggest that the delivery of anticancer agents with ASSHN-Lip could be useful for targeted cancer therapy. 3209 VTPNDDTFDPFR[38.7 ± 5.4] RPLDLYPGSGQE[85.8 ± 8.3] 2 Phage display (common panning) 3-5 PMID:28475831 Anti-Fumonisins B1 monoclonal antibody Fumonisin B1 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Mimotopes, the phage-displayed peptides, A2 (VTPNDDTFDPFR) and D1 (RPLDLYPGSGQE), were tested in competitive ELISAs. Four-parametric logistic fit (OriginPro 9.0) was used to calculate the IC50 values (ng/ml). Clone A2, with peptide sequence VTPNDDTFDPFR, showed the best response in ELISA in terms of sensitivity and reproducibility and was selected for microarray development. A biotinylated synthetic derivative of this mimotope was immobilized onto epoxy-glass slides and fumonisin B1 was detected in a competitive binding inhibition assay using the antifumonisin antibody and a labelled secondary antibody. The array showed an IC50 value of 37.1 ± 2.4 ng mL-1 (n = 9), a detection limit of 11.1 ng mL-1, and a dynamic range from 17.3 to 79.6 ng mL-1. Good specificity towards fumonisin B1 and its structural analog, fumonisin B2, was observed, together with negligible cross-reactivity for other mycotoxins produced by the same fungi species. The mimotope microarray was applied to the analysis of fumonisin B1 in spiked maize and wheat samples. 3210 VHWDFRQWWQPS[1.75 ± 0.12] 1 Phage display (common panning) PMID:28572662 Polystyrene 96-well microtiter plate (polystyrene, PS) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of the positive phage clones to polystyrene were measured by phage ELISA. The OD450nm values at the concentration of TBS with 0.20% (w/v) NFM were measured. Data shown were re-generated from the Figure 2 in the reference. Peptide VHWDFRQWWQPS, PB-TUP, has been isolated on target proteins (folate receptor-α, programmed death 1 and programmed death-ligand 1). ELISA and phage recovery assay demonstrated that PB-TUP phage had a significant superior affinity to polystyrene solid surface compared with control phage clones. Propagation rate assays of the selected phage clones showed that the growth rate of PB-TUP phage was not superior to the control phages. Furthermore, the binding of PB-TUB to polystyrene was concentration dependent and varied with solution pH. Molecular modeling revealed that stable structures of α-helix and β-turn may contribute to the binding of PB-TUP to polystyrene plate. The PB-TUP sequence was fused to the N-terminus of peptide P2 and the fusion peptide significantly increased the binding affinity to polystyrene. The fusion peptide also enhanced the cell adhesion ability of peptide P2 with human umbilical vein endothelial cell (HUVEC). The addition of the polystyrene binding peptide provided a convenient method for peptide immobilization. 3211 SGNNPLHVHHDKR 1 Phage display (common panning) 5 PMID:28578326 T lymphocyte leukemia cell line Jurkat Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) By fusing the Jurkat-binding peptide (SGNNPLHVHHDKR) to the C-terminus of IFNα, we constructed an engineered chimeric IFNα molecule (IFNP) for the treatment of adult T-cell leukemia/lymphoma (ATL). We found that IFNP exhibited significantly higher activity than wild type IFNα in inhibiting the growth of leukemia cells and inducing cell blockage at the G0/G1 phase. The synthetic IFNP molecule exerted its antitumor activity by upregulating the downstream genes involved in the STAT1 pathway and in apoptosis. Using a cell receptor binding assay, we showed that this Jurkat-binding peptide facilitated the binding affinity of IFNα to the cell surface type I IFN receptor. The engineered IFNP molecule may prove to a novel antitumor approach in the treatment of patients with ATL. 3212 CNYCRLNLW(19)[0.36 ± 0.07] CRSTLQHSC(1)[0.27 ± 0.05] CLKNQSDQC(1)[0.20 ± 0.04] CSTSSRTGC(1)[0.04 ± 0.01] CMHVRHGLV(1)[0.16 ± 0.04] CDSAPTRKC(1)[0.16 ± 0.04] 6 Phage display (subtractive panning) 3 PMID:28584144 Nitrite reductase AniA Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Phage clones were purified and tested separately to measure their affinity to sAniA in the phage ELISA assay. Absorbance at 450 nm was measured. Readings were compared to wells, which underwent identical treatment but lacked sAniA (control) and to signal from a wild type phage, M13KE (of 1010 PFU), that does not display peptides (wild type). The mean and SEMs from seven independent experiments are shown. Data shown were re-generated from Figure 3C in the reference. Magnetic Ni-NTA bead based affinity capture was used to immobilize sAniA. A preclearance step was to remove Ni2+ and plastic binders from the phage pool. 3213 AYPPNLWQKALA(2)[0.21 ± 0.04] HLNHNDNYLPP(1)[0.14 ± 0.04] AYSDRIPSLWDT(1)[0.07 ± 0.04] GYIERGLALWNS(1)[0.50 ± 0.10] YNFDVWKDHWIY(1)[0.24 ± 0.08] KHYYGGDTTTLW(1)[0.22 ± 0.06] SYNFDLWSFGLE(1)[0.68 ± 0.03] HYDRGNLWFQKS(1)[0.17 ± 0.06] GYKVPLWSKPEY(1)[0.11 ± 0.03] AQYERFDWASWW(1)[0.16 ± 0.04] YPSTLREMFWH(1)[0.35 ± 0.07] THDFSKARLWPS(1)[0.36 ± 0.06] EYSSFRLWNIYT(1)[0.53 ± 0.08] YHDLNLWELNVR(1)[0.34 ± 0.06] YHPNGMNPYTKA(1)[0.50 ± 0.08] DYSKMRLWDLRP(1)[0.14 ± 0.04] SYKYPLWHSITL(1)[0.22 ± 0.04] SLSPHYMNLWNA(1)[0.29 ± 0.06] YTGRAIDLWTEW(1)[0.24 ± 0.06] QYPYDLWSSMQR(1)[0.43 ± 0.09] SMSYKDREMQMW(1)[0.21 ± 0.04] AYSEWYASLWDF(1)[0.53 ± 0.09] ASIYYKDRSLLW(1)[0.53 ± 0.11] 23 Phage display (subtractive panning) 3 PMID:28584144 Nitrite reductase AniA Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Phage clones were purified and tested separately to measure their affinity to sAniA in the phage ELISA assay. Absorbance at 450 nm was measured. Readings were compared to wells, which underwent identical treatment but lacked sAniA (control) and to signal from a wild type phage, M13KE (of 1010 PFU), that does not display peptides (wild type). The mean and SEMs from seven independent experiments are shown. Data shown were re-generated from Figure 3C in the reference. Magnetic Ni-NTA bead based affinity capture was used to immobilize sAniA. A preclearance step was to remove Ni2+ and plastic binders from the phage pool. 3214 HSVKPVVNLILR(34) HSIRLHTYPHMK(19) YSLRADSRWMPS(18) HSLRPEWRMPGP(15) HSIRTYWQSAQP(10) HSLKPSWLLLGY(2) HSLKPSLKQLAI(2) HSLREDWTLRMQ(2) APPGNWRNYLMP(2) TMGFTAPRFPHY(2) TMHSLRPEWRMP(1) HSVKPDWAQMLR(1) HSVKHDFRLLTK(1) HSIRSSHLHMFT(1) SGHQLLLNKMPN(1) APRLPQSLLPQL(1) SHALPLTWSTAA(1) APPMSRQSFDGV(1) NFMESLPRLGMH(1) LLADTTHHRPWT(1) MEGQYKSNLLFT(1) NTELTSYGPPPA(1) H-S-[ILV]-[KR],[KR]-X-P, [KR]-X-X-P 22 Phage display (subtractive panning, competitive panning) 4 PMID:28415614 Gap junction alpha-5 protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A phage library was first pre-cleared in an uncoated well, and then presented to the bait protein. Low-affinity binders were first eluted using a solution of free bait protein in TBS. One of the retrieved peptides (HSLRPEWRMPGP) showed a 58.3% homology with amino acids 5-to-16 of IκBα, a member of the protein complex inhibiting NFκB activation. Binding of IκBα (peptide) and Cx40 was confirmed by crosslinking and en face proximity ligation assay on carotid arteries. 3215 GASESYL 1 Phage display (common panning) PMID:28208072 Anti-triosephosphate isomerase (TIM) polyclonal antibody Triosephosphate isomerase, TIM Not determined. Not determined. Ph.D.-7 phage display library (X7) A conformational epitope (A71G74S69D75T73F72V67) was confirmed by the phage display technology. 3216 CKKKAGSGC(57) CKELRDMQC(54) CDAPRSRRC(47) CKGKGRKGC(41) CGKKESPRC(40) CNKSMRNSC(39) CNKQARAKC(38) CERAQNGGC(37) CTGRDGKSC(36) CVNQVGKRC(35) CRKVKGDKC(30) CKKGLKGKC(20) CREGKGVRC(20) CKKKESSRC(18) CKGGKGARC(16) CRDTGEEKC(16) CKSEHVKKC(16) CKGKNSGRC(15) CKKSGGKKC(15) CKGAEEKKC(15) CREKRDNKC(14) CRSNMREKC(13) CKRDLSRRC(13) CKGSVNKRC(13) [RK]-x(2)-[RK], C-[RK]-x(5)-[RK]-C 24 Phage display (in vivo) 3 PMID:28468593 Human gastric cancer cell line MKN-45P Not determined. Not determined. Not determined. CX7C T7 phage display library For ex vivo biopanning, 1.0e10 pfu of CX7C phage naïve library was incubated with tumor and organs excised from the mice with MKN-45 IP tumors. For in vivo biopanning, amplified phage pool from previous round was IP injected to an MKN-45 tumor mouse and incubated for 1 h. After the termination, tumors and organs were collected, washed with DMEM, and transferred in LB-NP40. The rescued phage was amplified, pooled, and used for the subsequent round of selection. IP3 peptide with the sequence of CKRDLSRRC was identified by in vivo phage display on a mouse model of peritoneal carcinomatosis of gastric origin (MKN-45P), using high-throughput sequencing of the peptide-encoding region of phage genome as a readout. The IP3 peptide contains a hyaluronan-binding motif, and fluorescein-labeled IP3 peptide bound to immobilized hyaluronan in vitro. After intraperitoneal administration in mice bearing peritoneal metastases of gastric and colon origin, IP3 peptide homed robustly to macrophage-rich regions in peritoneal tumors, including poorly vascularized micro-tumors. Finally, we show that IP3 functionalization conferred silver nanoparticles the ability to home to peritoneal tumors of gastric and colonic origin, suggesting that it could facilitate targeted delivery of nanoscale payloads to peritoneal tumors. Collectively, our study suggests that the IP3 peptide has potential applications for targeting drugs, nanoparticles, and imaging agents to peritoneal tumors. 3217 CKKGLKGKC(225) CVNQVGKRC(209) CRDTGEEKC(159) CTGRDGKSC(123) CERAQNGGC(84) CKELRDMQC(45) CKGKGRKGC(38) CKGKNSGRC(32) CREGKGVRC(28) CKSEHVKKC(20) CKGGKGARC(18) CKGAEEKKC(16) CKKKAGSGC(15) CNKQARAKC(15) CRSNMREKC(3) CGKKESPRC(2) CNKSMRNSC(2) CRKVKGDKC(2) 18 Phage display (in vivo) 3 PMID:28468593 Mouse colon adenocarcinoma cell line CT26 Not determined. Not determined. Not determined. CX7C T7 phage display library 3218 CKKKAGSGC(83) CKELRDMQC(80) CKKSGGKKC(77) CNKQARAKC(69) CKGGKGARC(62) CKGKGRKGC(56) CTGRDGKSC(55) CVNQVGKRC(50) CRKVKGDKC(36) CERAQNGGC(24) CNKSMRNSC(23) CRDTGEEKC(21) CREGKGVRC(20) CGKKESPRC(18) CKKGLKGKC(18) CKSEHVKKC(14) CKGKNSGRC(9) CKGSVNKRC(5) CREKRDNKC(4) CRSNMREKC(3) CDAPRSRRC(2) CKKKESSRC(2) CKGAEEKKC(2) CKRDLSRRC(2) 24 Phage display (in vivo) 3 PMID:28468593 Kidney Not determined. Not determined. Not determined. CX7C T7 phage display library 3219 CQNNNMTSC(47.4%) CNWGDRILC(31.6%) CTVRTSADC(15.8%) CIGNSNTLC(5.30%) 4 Phage display (subtractive panning) 5 PMID:28428053 Crohn disease (CD) inflamed mucosa Not determined. Not determined. Not determined. Ph.D.-7 and Ph.D.-C7C phage display library pool A subtraction step was performed initially by clearing non-specific binders against a freshly harvested normal colon mucosa of healthy volunteers undergoing colonoscopy. We identified one peptide (TVRTSAD) able to recognize selectively biopsies of inflamed mucosa of Crohn disease (CD) patients. 3220 CNWGDRILC(31.3%) CQNNNMTSC(31.3%) CIGNSNTLC(12.5%) CWPANTLKC(12.5%) 4 Phage display (subtractive panning) 5 PMID:28428053 Ulcerative colitis (UC) inflamed mucosa Not determined. Not determined. Not determined. Ph.D.-7 and Ph.D.-C7C phage display library pool A subtraction step was performed initially by clearing non-specific binders against a freshly harvested normal colon mucosa of healthy volunteers undergoing colonoscopy. We identified one peptide (WPANTLK) able to recognize selectively biopsies of inflamed mucosa of ulcerative colitis (UC) patients. 3221 CTNPGVHIC(54.5%) CLKLGEKWC(18.2%) 2 Phage display (common panning) 5 PMID:28428053 Normal colon mucosa of healthy volunteers Not determined. Not determined. Not determined. Ph.D.-7 and Ph.D.-C7C phage display library pool 3222 GAMHLPWHMGTL(3/13) GAMHLSWHMGTH(1/13) DPMHNNWHSSPI(1/13) GLDHLWWSSQTP(1/13) NPWEEQGYRYSM(1/13) NPWNEMWFQTSR(1/13) NNPWREMMYIEI(1/13) WADMMTSVTPWL(1/13) SEFPRSWDMETN(1/13) QHYETLAFRPKH(1/13) ATYNSVNRHSAV(1/13) 11 Phage display (common panning) 4/5 PMID:28529640 Huamn large cell carcinoma (LCC) cell line H460 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Three targeting phages (HPC1, HPC2, and HPC4) and their respective displayed peptides (GAMHLPWHMGTL, NPWEEQGYRYSM and NNPWREMMYIEI) were able to bind to both SCLC and NSCLC cell lines, as well as clinical specimens, but not to normal pneumonic tissues. In vivo optical imaging of phage homing and magnetic resonance imaging (MRI) of peptide-SPIONs revealed that HSP1 was the most favorable probe for multimodal molecular imaging. Using HSP1-SPION, the T2-weighted MR signal of H460 xenografts was decreased up to 42%. In contrast to the tight binding of HSP1 to cancer cell surfaces, NNPWREMMYIEI was preferentially endocytosed and intracellular drug delivery was thereby effected, significantly improving the therapeutic index of liposomal drug in vivo. Liposomal doxorubicin (LD) conjugated to GAMHLPWHMGTL, NPWEEQGYRYSM or NNPWREMMYIEI had significantly greater therapeutic efficacy than non-targeting liposomal drugs in NSCLC (H460 and H1993) animal models. Combined therapy with an NNPWREMMYIEI-conjugated stable formulation of liposomal vinorelbine (sLV) further improved median overall survival (131 vs. 84 days; P = 0.0248), even in aggressive A549 orthotopic models. Overall, these peptides have the potential to guide a wide variety of tailored theranostic agents for targeting therapeutics, non-invasive imaging, or clinical detection of SCLC and NSCLC. 3223 DYDRYSSALIAA(5)[0.42] GQPKTFLSVSEL(5)[0.14] YWTPGYTPSQTE(1)[NA] NNSRDTALRWFV(1)[NA] 4 Phage display (common panning) 3 PMID:28469983 Anti-HO-1 polyclonal antibody Heme oxygenase-1, HO-1 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The selection of positive phage clones specific to polyclonal anti-△rHO-1 antibodies was detected by competitive ELISA. The color intensity was determined spectrophotometrically at A450. Data shown were reproduced from Figure 3 in the reference. NA denotes not applicable. Screening of a phage display library revealed four epitopes that could interact with the polyclonal antibody prepared by immunizing rabbits with the purified HO-1 protein. Two of these four epitopes (DYDRYSSALIAA and GQPKTFLSVSEL) are responsible for HO-1 catalytic activity because their antibodies were able to neutralize HO-1 activity. 3224 KCCFPAQ(13) SILPYPY(4) YPAPWPP(3) QPWPTSI(3) WPTPPYA(1) MHAPPFY(1) 6 Phage display (subtractive panning) 5 PMID:28012848 Human colorectal cancer cell line HT29 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Three rounds of biopanning with a total of ~1e9 plaque-forming units (pfu) was performed with ~1e6 human normal intestinal cells Hs738.St/Int (control) to remove non-specific binders. The supernatant containing the cleared phage pool was amplified to ~2e11 pfu for positive selection against HT29 cells. We identified the peptide sequence KCCFPAQ, and measured an apparent dissociation constant of kd = 72 nM and an apparent association time constant of k = 0.174 min–1 (5.76 min). During fluorescence imaging of patients during endoscopy, regions of SSA had 2.43-fold higher mean fluorescence intensity than that for normal colonic mucosa. Fluorescence labeling distinguished SSAs from normal colonic mucosa with 89% sensitivity and 92% specificity. The peptide had no observed toxic effects in animals or patients. In the analysis of ex vivo specimens, peptide bound to SSAs had significantly higher mean fluorescence intensity than to hyperplastic polyps. 3225 NPVEDYLDYSVI ESYYMNPVEMFV LPTESNPVEDWI NPIESYIASVFS NPVELLLKTSSD NPVETQIVLSLV NPVEKWISMRTM NPVELLLLMGIS NGIERLLEEPVS NPVENWIDPKSI VNPVEYYLDTMR NPVEALLSKFHM NPVESYLANYTS VTNPYESLVQEK SPPEFSGSTVGL MVPEFSGSFPMR RMPPEFMGSLPQ QLFVPEFAGSSP LVPEFTGSTPFR SHGAIEFDGAFP VPEFAGHVPSTA VPEMNGSLIARK HPIVKTYFAQTT APAKTYFGQTTD QMKAWFPQTTYD ATKVMFPQRIYV QPAKTYFNQVTL QAAAKTMFPQNT SPSIDAFETSIF SWVLTATETGSS EMRFPSLSASDT QAPTLDAQETAL TALDAVSTGFSW GAIGSLTADSTS SLGSLSAYETGR TLYRPPLTSAET LIADLNAESTSR SLGAAGARVTTV ASGPLHAGATGL GHKPVLTALSTA VMPAIHAGVTGA VHPTLTVTSKEI ELDVSLRVMPKV SKLAMEIMSGPV SNAVTSSKSPRM QADATVLTKPKT KAVSDASRGTVF DYPKIANFQEYA GGQVRSIHSGPT QHWPTNVDSVTV ETKSDDMLLSNV MTVDRTVRVASK DLLIRDAITDTK AYHVDTVSDAGW VDTINLPQNTIQ VMSVNASTTAAN TLHAPMTIRSGP TNLHRVMTVVNM LRPNAVQTDTLA QMRQLTEYGSEK VLSSTAIKVDSV VHTVHDVFTAFG AKIRMFLDTDYK ASWGPIAIDRVN SQQYALTNSTTN QPQTKSFYPQYV NVVDRVNRTGVV WSALPERTSLPV RPAIVDQVSSSP QWNWRVRSVANV AQLHPTTLVKHK TSYRPPLNVCQD QSHSLFYPHPYG N-P-V-E-x(3), X-P-E-F-X-G-S-x(2), K-x(2)-F-P-Q-x-T 73 Phage display (common panning) 3 PMID:28167275 Human whole serum Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) We synthesize identified peptides in the array format and rescreen the serum used for phage panning to validate antibody binding peptides. Finally, we systematically vary the sequence of validated antibody binding peptides to identify those amino acids within the peptides that are crucial for binding “their” antibody species. The resulting immune fingerprints can then be used to trace them back to potential antigens. 3226 FIPFDPMSMRWE[0.0057,NT] SYIEPPERHRHR[0.0230,57] DDAKSRQGPLFR[0.0062,NT] NVNEGKAGVTGW[0.0036,NT] 4 Phage display (subtractive panning) 5 PMID:27989733 Myeloperoxidase Anti-human myeloperoxidase antibody Not determined. Not determined. Ph.D.-12 phage display library (X12) Surface plasmon resonance (SPR),Quartz crystal microbalance (QCM) A quartz crystal microbalance sensor (QCM D-300; Q-sense, Sweden) equipped with a QSX 301 gold chip was used to determine the molar binding ratio of hMPO to the immobilized ligand, i.e. the anti-hMPO antibody or the synthesized peptide. To study the binding kinetics and equilibrium, a surface plasmon resonance (SPR) biosensor (Biacore 3000, GE Healthcare, USA) equipped with a CM5 Sensor chip was used. The first colum of the affinity values represents the molar binding ratio determined by QCM, the second column denotes the Kd values (nM) determined by SPR. Before proceeding to the next round of biopanning, non-specific phages that were included in the selected phages were removed by a ‘subtraction’ (or negative selection) step by using epoxy-functionalized magnetic particles without a myeloperoxidase from human leukocytes (hMPO) coating (‘nude’ magnetic particles). The supernatant from this subtraction step was introduced to the next round. We selected and chemically synthesized a 12-mer peptide (SYIEPPERHRHR). Quartz crystal microbalance and surface plasmon resonance analyses revealed that the molar binding equilibrium ratio of the synthesized peptide was 0.023, approximately 43-fold lower than that of the anti-hMPO antibody. The dissociation constant (Kd) was 57 nM, which was comparable to that of the native antibody (83 nM). We biotinylated the peptide at its N-terminus and attached the biotinylated peptide to the surface of streptavidin-coated magnetic particles to assess its ability to selectively capture hMPO. The binding equilibrium data were similar to the previous analyses; specifically, around 0.021 mol peptide bound to 1 mol of hMPO. Antigen capture was found to be selective and to be relatively little influenced by the presence of human serum albumin (HSA), an abundant constituent of serum. 3227 HPNYDHDRMHTQ(16/50)[0.068] SMQYADHPVNTG(13/50)[0.272] FRVIYTDMPGAH(9/50)[0.250] 3 Phage display (common panning) 4 PMID:28346102 Pancreatic secretory granule membrane major glycoprotein GP2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Saturation equilibrium assay To measure the binding strength of the selected peptide ligands to GP-2 (Kd, μM), saturation binding assays were performed. Binding affinity of each peptide was calculated by nonlinear regression and transformed to Scatchard plot using the GraphPad Prism 6 program (GraphPad Software, La Jolla, CA). Four rounds of biopanning were performed using a 96-well plate (Corning, Corning, NY). Selected peptides were conjugated to the C-terminal of enhanced green fluorescence protein (EGFP) and evaluated for their ability to induce an immune response in mice. One of our selected peptides, Gb-1 (HPNYDHDRMHTQ), showed high binding affinity to GP-2 and, when fused to EGFP, significantly increased the uptake of EGFP by M cells compared to EGFP alone. After oral administration, the Gb1-EGFP fusion induced efficient mucosal and systemic immune responses in mice measured at the level of antigen-specific serum and fecal antibodies, cytokine secretion, and lymphocyte proliferation. Furthermore, the IgG subclasses and cytokine secretion showed that ligand Gb-1 induced a Th2-type immune response. 3228 CAVECPSTLDKH[5.05, 4.35] LSSNPKLYVGLE[5.32, 4.42] 2 Phage display (subtractive panning) 2 PMID:28434129 Human IgG anti-HEV positive serum Capsid protein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The sandwich phage ELISA was used to test mimotope reactivity against anti-HEV positive sera S51 and S56, respectively. The ratio of clone OD absorbance and wt (wild-type phage M13KE) OD absorbance at 450 nm was calculated. The first column of the affinity values is the ratio with S51, the second column is the ratio with S56. The positive selection was carried out with protein G magnetic micro beads (Miltenyi Biotec) labelled with human IgG anti-hepatitis E virus (HEV) positive serum. The eluted bound phage was negatively selected with beads labelled with human IgG anti-HEV negative serum. Unbound phage from negative selection was amplified in E. coli ER2738 by a high-throughput method [38] and subjected to one more round of biopanning with different positive and negative sera. Four mimotopes did not have homology with the primary sequence of HEV ORF2 capsid but competed effectively with a commercial HEV antigen for binding to anti-HEV reference serum. When the reactivity profile of each mimotope was compared with Wantai HEV-IgG ELISA, the most sensitive HEV immunoassay, mimotopes showed 95.2–100% sensitivity while the specificity ranged from 81.5 to 95.8%. PepSurf algorithm was used to map affinity-selected peptides onto the ORF2 crystal structure of HEV genotype 3, which predicted that these four mimototopes are clustered in the P domain of ORF2 capsid, near conformational epitopes of anti-HEV neutralising monoclonal antibodies. These HEV mimotopes may have potential applications in the design of structural vaccines and the development of new diagnostic tests. 3229 HCSSAVGSWTWENGKWTWKGIIRLEQ[65] CSSPIQGSWTWENGKWTWKGIIRLEQ[16] HSCSSPIQGSWTWENGKWTWKGIIRLEQQP[3] 3 Phage display (common panning) 4 PMID:22271427 Fibronectin extradomain B, EDB Not determined. Not determined. Not determined. Aptide pIGT2-based phage display library (X6GSWTWENGKWTWKGX6) Surface plasmon resonance (SPR) Kinetic binding data (Kd, nM) for selected aptides, as determined by affinity measurements using BIAcore chips adapted for surface plasmon resonance. To immobilize the target protein (biotin-fibronectin extradomain B), streptavidin was immobilized directly on 96-well plates overnight at 4°C, and then the target protein was immobilized on the plate-bound streptavidin. 3230 HANFFQGSWTWENGKWTWKGWKYNQS[30] 1 Phage display (common panning) 4 PMID:22271427 Vascular endothelial growth factor A, VEGF-A Vascular endothelial growth factor receptor 1, VEGFR-1 1FLT,1QTY, Not determined. Aptide pIGT2-based phage display library (X6GSWTWENGKWTWKGX6) Surface plasmon resonance (SPR) Kinetic binding data (Kd, nM) for selected aptides, as determined by affinity measurements using BIAcore chips adapted for surface plasmon resonance. To immobilize the target protein, streptavidin was immobilized directly on 96-well plates overnight at 4°C, and then the target protein was immobilized on the plate-bound streptavidin. 3231 ASTINFGSWTWENGKWTWKGYTRRWN[93] 1 Phage display (common panning) 4 PMID:22271427 T-cell antigen CD7 Not determined. Not determined. Not determined. Aptide pIGT2-based phage display library (X6GSWTWENGKWTWKGX6) Surface plasmon resonance (SPR) Kinetic binding data (Kd, nM) for selected aptides, as determined by affinity measurements using BIAcore chips adapted for surface plasmon resonance. To immobilize the target protein, streptavidin was immobilized directly on 96-well plates overnight at 4°C, and then the target protein was immobilized on the plate-bound streptavidin. 3232 CSQNAYGSWTWENGKWTWKGWLYQNF[71] 1 Phage display (common panning) 4 PMID:22271427 His6-tag Not determined. Not determined. Not determined. Aptide pIGT2-based phage display library (X6GSWTWENGKWTWKGX6) Surface plasmon resonance (SPR) Kinetic binding data (Kd, nM) for selected aptides, as determined by affinity measurements using BIAcore chips adapted for surface plasmon resonance. To immobilize the target protein, streptavidin was immobilized directly on 96-well plates overnight at 4°C, and then the target protein was immobilized on the plate-bound streptavidin. 3233 PD(93.33%) DT(6.67%) 2 Phage display (common panning) 3 PMID:28548828 Gold nanocube Not determined. Not determined. Not determined. pVIII AE(X)2DP phage display library (AEX2DP) This clone displayed Pro–Asp on the surface of the bacteriophage and had amplification yields similar to those of the wild-type helper bacteriophage (VCSM13). The clone displayed even binding of gold nanocubes along its length and minimal aggregation after binding. We conclude that, for efficient amplification, the exposed pVIII amino acid length should be limited to six residues and Ala–Glu–Pro–Asp–Asp–Pro (AEPDDP) is the ideal fusion protein sequence for guaranteeing the optimal formation of a complex with gold nanocubes. 3234 SDG DID DVS ESQ ENS RMG DNA EVP 8 Phage display (common panning) 3 PMID:28548828 Gold nanocube Not determined. Not determined. Not determined. pVIII AE(X)3DP phage display library (AEX3DP) 3235 AGDS 1 Phage display (common panning) 3 PMID:28548828 Gold nanocube Not determined. Not determined. Not determined. pVIII AE(X)4DP phage display library (AEX4DP) 3236 FRGDKMQL(7) HWFWSKWW(5) WWKFMWFQ(1) RAWWSHWH(1) WRWASWFW(1) WHQNIHSP(1) 6 Phage display (common panning) 12 PMID:28468949 Head and neck squamous cell carcinoma (HNSCC) cell line HNO97 Not determined. Not determined. Not determined. SFTI-based phage display (SFTI8Ph) library (X8) Initially, phages were incubated for 1 hour with HNO97 grown to 90% confluence. Unbound phages were removed by washing steps and the cells were lysed with 1% Triton X-100 solution. Phages isolated from cell lysate were amplified and packaged in XL1 blue bacteria overnight and precipitated in polyethylenglycole solution . Subsequently, the phages were exposed alternatingly to HNO97 cells and HNO97 protein fractions (100nmol/L) in96-well plates for1 hour. After 5 PBS washing steps phages were eluted in 100 mLGlycin/HCl (pH 2.2) per well, neutralized by 15 mL Tris-HCl(pH 9.1) and amplified in XL1 blue bacteria. For titration, the phages were diluted and grown on agar plates. Twelve selection rounds were performed followed by single stranded DNA isolation of 16 clones (QIApreo Spin M13 Kit;Qiagen). 3237 DWYRDPVFDVLL LWRRVVVDSYGV YMWIACPDGWDC WWDWAVQSETLL WWVNPLGVTQDV EYPWWMEQFYLE WWAEAWFAWTPG ISWHSDWWVELV 8 Phage display (common panning) 4 PMID:28701344 The μ (mu) homology domain of suppressor of yeast profilin deletion Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) From the screen, we identified eight potential Syp1 ligand sequences and, to aid in the search for potential motifs, we compared the peptide sequences to that of Mid2 (251-316), the smallest fragment of Mid2 previously shown to bind the Syp1 μHD. Two DxY sequences (DWYRDPVFDVLL and LWRRVVVDSYGV), two YxxΦ motifs (YMWIACPDGWDC and EYPWWMEQFYLE), and several instances of two consecutive residues with bulky side chains, including one WY (DWYRDPVFDVLL) and five WW sequences (WWDWAVQSETLL, WWVNPLGVTQDV, EYPWWMEQFYLE, WWAEAWFAWTPG and ISWHSDWWVELV), were present in the peptides. All of these potential Syp1-interacting motifs are present in Mid2 (251-316). 3238 NPNSAQG 1 Phage display (common panning) 3 PMID:28710679 Cellular tumor antigen p53 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA ELISA of phage against p53 domains was carried out by coating 100 µl of 10 µg/ml of peptide solution (in 2% non-fat dry milk in 1x PBS) overnight at 4 °C. Reaction is read using a microplate reader set at 410 nm. The estimated gain of signal (% phage/control) for the phage N6 (with the sequence of NPNSAQG) is 48.23%. Three panning rounds were performed directly on the fusion GST-p53 that was previously fixed on G-Sepharose beads. Phages were sequenced and shown to contain a consensus sequence NPNSAQG 3239 QVNGMLGERSQQ(5)[0.77, 0.21] VYPYPKVDLSQD(3)[0.64, 0.28] DVSSLYDGPVGD(2)[0.64, 0.27] HAQGSLGTPVFM(2)[0.83, 0.24] Y-x(2)-P-x-G-D-L-G 4 Phage display (common panning) 3 PMID:28711695 Anti-FMDV monoclonal antibody 6C9 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Four phage clones were selected after three rounds of biopanning, and their binding activity to MAb 6C9 was analyzed in phage ELISA. Murine MAb 5D12 and wild-type M13 phage (PL) were used as negative controls. For each clone, the absorbance at 450 nm was calculated as an average of three duplicates. Data shown were absorbance values (the first column of the affinity values) reproduced from Figure 2. Additionally, binding inhibition of phage clones and whole virus antigens to MAb 6C9 were analyzed in competitive ELISA. TBS was used as a negative control. For each clone, the absorbance at 450 nm was calculated as the average of three duplicates. Data shown were absorbance values (the second column of the affinity values) reproduced from Figure 3. The M13 phage display library displaying 12-mer random peptides (Ph.D-12 Phage Display Peptide Library Kit, New England Biolabs) was used. Affinity selection of the phage clones from the random peptide library was conducted following the manufacturer's recommendations with minor modifications. Screening of a phage-displayed random 12-mer peptide library revealed that MAb 6C9 bound to phages displaying the consensus motif YxxPxGDLG. 3240 DMHADHAVYTPR(13) DMHAEHPGYRTA(1) NIHPDPPVYTPR(1) NMHADHPGYRTR(1) DMHADHPGYRTG(1) DMHAEHPGDRTG(1) DMLADHPGDRTR(1) DMHADHPGDRTR(1) 8 Phage display (competitive panning) 5 PMID:28715634 Tumor necrosis factor receptor superfamily member 12A Tumor necrosis factor ligand superfamily member 12 Not determined. Not determined. Ph.D.-12 phage display library (X12) Considering the possible non-specific binding that emerges in the interaction between the phage library and L-Fn14 CRD, competitive elution was conducted to avoid the elution of background-bound phage. The natural ligand of Fn14, TWEAK, was served as the eluent. After five rounds of panning, twenty plaques were randomly selected and the amplified DNA was sequenced, of which thirteen binding clones gave rise to the following sequence: DMHADHAVYTPR. 3241 LANNMLTLSPKMPGGGYW(3) SPTIFSTLSPKMPGGGYW(3) AEAFAFTLSPKMPGGGYW(1) ATTLTTTLSPKMPGGGYW(1) AGDLQETLSPKMPGGGYW(1) ASSEIHTLSPKMPGGGYW(1) ASLPLMTLSPKMPGGGYW(1) SQAAMLTLSPKMPGGGYW(1) MTTTNETLSPKMPGGGYW(1) MFPANATLSPKMPGGGYW(1) SPIVPDTLSPKMPGGGYW(1) SPLLSSTLSPKMPGGGYW(1) VPQGNXTLSPKMPGGGYW(1) VPQGATTLSPKMPGGGYW(1) LPYTIPTLSPKMPGGGYW(1) GPHTLATLSPKMPGGGYW(1) DLLHWTTLSPKMPGGGYW(1) GSAAPETLSPKMPGGGYW(1) LPLAPYTLSPKMPGGGYW(1) TQLAASTLSPKMPGGGYW(1) SNSQAYTLSPKMPGGGYW(1) HLATPTTLSPKMPGGGYW(1) HNPIPYTLSPKMPGGGYW(1) 23 Phage display (in vivo) 3 PMID:28658990 The middle ear Not determined. Not determined. Not determined. X6TLSPKMPGGGYW M13 bacteriophage library (X6TLSPKMPGGGYW) 50ul aliquots of each 18-mer phage library containing approximately 50 copies of each distinct 18-mer phage peptide were applied to the external surface of intact TMs of six rats. The phage samples were then pooled, amplified in E. coli, and the resulting library applied to the TMs of additional rats for two more rounds of library enrichment. After the third round, 30 individual phage plaques were selected at random and sequenced to identify the expressed peptide. After a 15-minute incubation on the TM, the external ear canal was repeatedly rinsed with PBS to remove all bacteriophage. The rats were then sacrificed and the ME bullas harvested intact. The exterior surface of the bulla was then extensively rinsed to remove contaminating phage. The bulla was then opened, exposing the ME contents. Fluid was collected from the ME, diluted in PBS and bacteriophage were titered using E. coli ER2738 (New England Biolabs, Cambridge, MA). The phage samples were then pooled, amplified in E. coli, and the resulting library applied to the TMs of additional rats for two more rounds of library enrichment. Transport of phage bearing peptide TMT-3(18)-1 was 8.9-fold higher than that of the TMT-3 12-mer. Phage expressing peptide TMT-3(18)-2 exhibited a 3-fold enhancement, while the TMT-3(18)-3 phage exhibited a significant decrease. 3242 KISVYRSADSTKTTHLTL(26) TEVTSRSADSTKTTHLTL(3) EGNLFPSADSTKTTHLTL(1) 3 Phage display (in vivo) 3 PMID:28658990 The middle ear Not determined. Not determined. Not determined. X6SADSTKTTHLTL M13 bacteriophage library (X6SADSTKTTHLTL) 50ul aliquots of each 18-mer phage library containing approximately 50 copies of each distinct 18-mer phage peptide were applied to the external surface of intact TMs of six rats. The phage samples were then pooled, amplified in E. coli, and the resulting library applied to the TMs of additional rats for two more rounds of library enrichment. After the third round, 30 individual phage plaques were selected at random and sequenced to identify the expressed peptide. After a 15-minute incubation on the TM, the external ear canal was repeatedly rinsed with PBS to remove all bacteriophage. The rats were then sacrificed and the ME bullas harvested intact. The exterior surface of the bulla was then extensively rinsed to remove contaminating phage. The bulla was then opened, exposing the ME contents. Fluid was collected from the ME, diluted in PBS and bacteriophage were titered using E. coli ER2738 (New England Biolabs, Cambridge, MA). The phage samples were then pooled, amplified in E. coli, and the resulting library applied to the TMs of additional rats for two more rounds of library enrichment. Transport of phage bearing peptide TMT-3(18)-1 was 8.9-fold higher than that of the TMT-3 12-mer. Phage expressing peptide TMT-3(18)-2 exhibited a 3-fold enhancement, while the TMT-3(18)-3 phage exhibited a significant decrease. 3243 FRLGLLKAFRRLF[210] RRDMCSMYGLC[NT] CDYFALGWRCWL[NT] 3 Phage display (competitive panning) 4 PMID:28676225 CTP synthase 1 Not determined. Not determined. Not determined. T7 phage display library Surface plasmon resonance (SPR) SPR experiments were performed on a BiacoreT100 instrument equipped with SA sensor chips (GE Healthcare). Binding affinities (Kd) were calculated from the equation Kd = koff/kon. We evaluated the binding activity of synthetic peptides by SPR analysis. CTpep-3 (FRLGLLKAFRRLF) showed CTPS1-binding in a peptide concentration dependent manner, and the Kd value was estimated as 210 nM. At the 3rd and 4th rounds, phages were subtracted using cytidine triphosphate synthase 1 (CTPS2) immobilized magnetic beads. 3244 SADSTKTTHLTL(10/22) SPPGKFLESLRS(6/22) DVGAGRWFSDNG(4/22) TLSPKMPGGGYW(2/22) SDDSRPIAQFAI(NA) 4 Phage display (in vivo) 3 PMID:26946957 The middle ear Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 50 μl aliquots of the naïve phage library, each containing 1,000 copies of the 2.9× 109 different peptide sequences in the library, were placed on the outer surface of the TM (Tympanic membrane) of anesthetized and immobilized rats with active OM (Otitis media), then laid on their side for 1 hr. Te remaining phage solution was removed by wicking and the TMs were then extensively washed with PBS several times to remove any unbound phage particles from the external canal and reduce the possibility of ME (Middle ear) sample contamination. The animals were then sacrificed by decapitation under deep anesthesia to isolate the ME from the single side where phage was applied. The bullas of the rats were then dissected, isolated, and further rinsed in PBS prior to opening, as another precaution against contamination. The bulla then was opened and ME effusion was collected in a sterile tube for further analysis. The number of phage particles present in the ME fluid sample was assessed using an aliquot of supernatant in standard plaque assay. Phage in the remaining fluid were amplified by infecting bacteria via standard methods29 to generate a library of candidate TM-translocating phage, which was re-applied onto the TMs again using the protocol described above. The screen was successively repeated twice before DNA sequencing. There was a strong selection for a central positively charged residue, lysine or arginine (K/R), in all five peptides. 3245 TPMVERNYNAAD(5/30) ALWPPNLHAWVP(3/30) THPSTKVPGTPA(3/30) TFNPPPPQMPST(3/30) ILVGDESDGAKP(2/30) MSTPLGQYTGTK(2/30) HYPTTQLPHHKQ(2/30) VQNSTLSTKMDS(2/30) MVEKYATLFQSS(1/30) SHQNYHLTPQPP(1/30) APTTQRVGWDQS(1/30) AHSANNFDVKGI(1/30) MPLMSEPALEML(1/30) NYEFSISEQSHD(1/30) VNPSSLHSHNHS(1/30) ATNVVIMTQPRP(1/30) 16 Phage display (in vivo) 10 PMID:26946957 The middle ear Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the first stage, the commercial phage library was applied onto the TM (Tympanic membrane) of ears with OM, allowed to incubate for 1 hour (hr), and the external canal was extensively rinsed with PBS to remove unbound phage as described earlier. The TM was then harvested from the ME (Middle ear) bulla, washed with PBS, and homogenized. Phage present in the homogenate was amplified, and the process was repeated twice to generate a library enriched for phage with TM-binding activity. We then exposed TMs of infected rats to this preselected library, followed by elution with low pH buffer to remove all phage that had not internalized into the membrane. The TM was again harvested by dissection, homogenized, and phage libraries were amplified in 3 consecutive rounds. The resulting “internalizing phage” selected library was again applied to the TM surface of ears for 1 hr in rats with OM (Otitis media). Fluids were then recovered from the opposite ME cavity side, phage within the samples were amplified, and re-applied for three additional rounds. 30 phage particles from the final two rounds of in vivo selection were chosen at random and sequenced to identify the peptides displayed by the transported phage. Using this successive selection procedure (ten rounds total), additional populations of peptide-targeted phage with the ability to bind, internalize and finally transit the TM were identified. Two animals per condition were used. Pooled samples recovered from the final round of in vivo selection were sequenced. Sorting the sequences into groups using ClustalW2 for alignment identified 5 major phylogenetic groups. Group 1 contained two peptides that are characterized by a conserved NxSTLSTK core motif. Group 2 had the noteworthy PPxx core motif and a variable C-terminal region. These were different from group 3 which appeared to have a PT(S/T)E(R/K) central motif preceding a variable C-terminal region. The fourth category of sequences contained a polar negatively charged N-terminal region followed by the consensus aliphatic motif SISEQxR though the C-terminal region. The final sequence group contained an interesting HxxYxxD motif and a C-terminal QxP motif. These results indicate that the different grouped peptides could be binding a variety of targets due to the differences in the consensus motifs and the physical and chemical characteristics of the amino acid groups involved. Many of the residues contained are polar, either negatively or positively charged, which suggests that electrostatic interactions are at play. 3246 DSQFNKYSIATV(2/19)[0.60] VQCRLGPPWCAK(1/19)[1.32] SLLHTSMPSMIA(1/19)[0.57] KVEPGTGAHSWQ(1/19)[0.55] VADSNWINPRGS(1/19)[0.52] QGPHEHTKIHID(1/19)[0.41] 6 Phage display (common panning) 3 PMID:28780344 Cholera toxin subunit B, CTX-B Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA ELISA signal was measured at 405nm using a microplate spectrophotometer (Multiskan FC, Thermo Scientific, Waltham, MA, USA). Binding affinity values shown were regenerated from Figure 1A. Biopanning over recombinant CTX-B led to rapid screening of a unique peptide with an amino acid sequence of VQCRLGPPWCAK, and the phage-displayed peptides analyzed using ELISA, were found to show specific affinities towards CTX-B. To address the use of affinity peptides in development of the biosensor, sequences of newly selected peptides were modified and chemically synthesized to create a series of affinity peptides.Performance of the biosensor was studied using plasmonic-based optical techniques: localized surface plasmon resonance (LSPR) and surface-enhanced Raman scattering (SERS). The limit of detection (LOD) obtained by LSPR with 3σ-rule was 1.89ng/mL, while SERS had a LOD of 3.51pg/mL. In both cases, the sensitivity was much higher than the previously reported values, and our sensor system was specific towards actual CTX-B secreted from V. cholera, but not for CTX-AB5. 3247 GVKCTWSSIVDWVCVDM(11)[7.3 ± 0.2] GTRCDWSAAYGWLCYDY(4)[NT] RSVCVWTAVTGWDCRND(1)[ 4.4 ± 0.1] x(3)-C-x-W-x(5)-W-x-C-x(3) 3 Phage display (subtractive panning) 4-5 PMID:28758361 Chicken egg yolk immunoglobulin, IgY Not determined. Not determined. Not determined. T7 X3CX9-10CX3 phage display library (X3CX9-10CX3) Surface plasmon resonance (SPR) Surface plasmon resonance (SPR) analysis was performed on a BIAcore T200 (GE Healthcare, Little Chalfont, UK). Binding kinetic parameters (Kd, μM) were calculated using BIAevalution Version3.2 Software (GE HealthCare). T7 phage libraries of X3CX7–10CX3 were incubated in wells coated with BSA to remove non-specific phage and then added to IgY-coated wells. The synthetic peptides (Y4-4: GVKCTWSSIVDWVCVDM, Y5-55: RSVCVWTAVTGWDCRND) showed high binding specificity to IgY-Fc and moderate affinity for IgY-Fc (Kd: Y4-4 = 7.3 ± 0.2 μM and Y5-55 = 4.4 ± 0.1 μM) by surface plasmon resonance analysis. To evaluate the ability to purify IgY, we performed immunoprecipitation and affinity high-performance liquid chromatography using IgYbinding peptides; the result indicated that these peptides can be used as affinity ligands for IgY purification. We then used Y4-4 peptide-conjugated column to purify IgY from egg yolks pre-treated using an optimized delipidation technique. 3248 CPFMSVANC(2) CAISSPGSC(1) CNAKHHPRC(1) CDSRWHSVC(1) CNGGYAFEC(1) CEHHKNTLC(1) CNSRLNQHC(1) CEIWSRAAC(1) CPGEPPTLC(1) CELQGWHLC(1) CPMFLASLC(1) CEPSLIPFC(1) CPRVNMLHC(1) CGENIQQYC(1) CQNHQSRHC(1) CGLNKTPGC(1) CRSDSYNKC(1) CGPKTPHKC(1) CSGALTSPC(1) CGSKLDALC(1) CSKQSTIAC(1) CHQWNSNSC(1) CSMEELVTC(1) CHTAHKATC(1) CSTEHTRSC(1) CKDQGQMRC(1) CTASPSGLC(1) CKPHTGHVC(1) CTDPNSTTC(1) CKPPGSYFC(1) CTNANSHLC(1) CLDSPNNVC(1) CTPDASRKC(1) CLKLTAASC(1) CTSNTKFNC(1) CLNDSVHQC(1) CTTYNATSC(1) CLNVPTKS(1) CWPPSTYSC(1) CLSDYSIMC(1) CYGKNNQRC(1) CMLNTSVAC(1) CYSELTGNC(1) 43 Phage display (common panning) 3 PMID:28757359 The human colon carcinoma Caco-2 cell Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Phage displaying cyclic heptapeptide DNPGNET (DNP-phage) showed the greatest permeability across a Caco-2 cell monolayer and mouse intestinal epithelium. Interestingly, DNPGNET (DNP) does not contain any basic amino acids. Its isoelectric point (pI) was estimated to be 2.72. It did not reduce the viability or tight-junction integrity of Caco-2 cells at concentrations up to 100 μM for 24h. Up take of either DNP phage or fluorescence-labeled DNP derivative (AC-DNPGNET-CGGGS modified with 5/6-FAM at the C-terminal; the cysteines serve to generate the cyclic peptide via disulfide bond formation, and GGGS is the phage linker) by Caco-2 cells was inhibited by low temperature, unlabeled AC-DNPGNET-CGGGS and EIPA, a macropinocytosis inhibitor. Thus, DNP appears to facilitate transcellular permeability of phages via macropinocytosis, but not paracellular diffusion. These findings indicate that DNP is a promising candidate as a carrier to promote intestinal absorption of macromolecular drugs. 3249 KVVKTHR[1.34] MVYLTEK[0.49] KTVLTHR[0.28] NAGHLSQ[0.20] NMHTPMV[0.18] 5 Phage display (subtractive panning) 4 PMID:28744632 Tumor necrosis factor ligand superfamily member 10 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance at 450 nm was measured on a Tecan microplate spectrophotometer (Genios). Data shown were regenerated from Figure 2b. For phage selection, phage particles were pre-incubated with mouse Fc. Then, bound phages were removed using magnetic beads with immobilized protein G (Invitrogen), while the supernatant was used for further selection of more specific peptides by adding mouse DR5-Fc. The cyclopeptide sequences selected from the library show appreciable homology at positions 2 (valine), 4 (leucine), and 5 (tyrosine). Substitution of valine for threonine in position 2 in a KVVLTHR peptide leads to a decrease in binding. It is possible that these very amino acids play the key role in interaction between the peptide and DR5. 3250 CLPTKFRSC(5)[high binding, 270 ± 151] CHEQQNPHC(1)[low binding, NT] CHKHKDSQC(1)[low binding, NT] CHLGEMGHC(1)[low binding, NT] 4 Phage display (common panning) 3 PMID:28733706 The extracellular domain of receptor tyrosine-protein kinase erbB-3 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance at 405 nm was read by the spectrophotometer. Peptide ELISA with HER3P1 (CLPTKFRSC) demonstrates an affinity of 270 ± 0.151 nM. The receptor tyrosine-protein kinase erbB-3 extracellular domain (R&D Systems, Minneapolis, MN) was conjugated with long chain biotin (Thermo Scientific, Waltham, MA) utilizing standard NHS ester chemistry and purified. Purified, biotinylated receptor tyrosine-protein kinase erbB-3 was bound to Dynabeads M-280 streptavidin beads (Thermo Scientific). The selection converged to a consensus peptide sequence that was subsequently found to bind receptor tyrosine-protein kinase erbB-3 (HER3) with an affinity of 270 ± 151 nM. The peptide with the sequence of CLPTKFRSC, termed HER3P1, was bound with high selectivity to HER3 over other similar receptor tyrosine kinases such as EGFR and HER2. Furthermore, HER3P1 was able to distinguish between high and low HER3-expressing cells in vitro. The peptide was radiolabeled with Ga-68 and demonstrated to specifically bind HER3 by in vivo PET imaging. Uptake of [ 68Ga]HER3P1 was highly specific for HER3-positive tumors, with tumor-to-background ratios ranging from 1.59–3.32, compared to those of HER3-negative tumors, ranging from 0.84–0.93. The uptake of [68Ga]HER3P1 also demonstrated high (P < 0.001) correlation with protein expression as quantified by Western blot and confirmed by biodistribution. HER3P1 accurately quantifies expression of HER3 by PET imaging and has potential utility as a clinical imaging agent. 3251 LPKQKRRQRRRM HASPPKNSHFAL LSNQSNPGIAKS LSKNRRNRRRQH WSFNWSMHSLGN KQRMQTSNRRRR NRRHRNLRRPPL QKRIRIQRRSLR QLRRRHRRIRTN RRHQSRILIQRS RQR 10 Phage display (common panning) 5 PMID:28860066 Mouse melanoma carcinoma cell line B16 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) MT23 (LPKQKRRQRRRM) showed higher penetration efficiency based on fluorescence microcopy and quantitative assay, and it has capability for mediating functional Apoptin into cells in vitro or in vivo. 3252 HSVKPVVNLILR(34) HSIRLHTYPHMK(19) YSLRADSRWMPS(18) HSLRPEWRMPGP(15) HSIRTYWQSAQP(10) HSLREDWTLRMQ(2) HSLKPSLKQLAI(2) TMGFTAPRFPHY(2) APPGNWRNYLMP(2) HSLKPSWLLLGY(2) HSVKPDWAQMLR(1) HSIRSSHLHMFT(1) HSVKHDFRLLTK(1) SGHQLLLNKMPN(1) APRLPQSLLPQL(1) SHALPLTWSTAA(1) APPMSRQSFDGV(1) NFMESLPRLGMH(1) LLADTTHHRPWT(1) MEGQYKSNLLFT(1) NTELTSYGPPPA(1) TMHSLRPEWRMP(1) H-S-[ILV]-[KR], x-P-[KR], x(2)-P-[KR] 22 Phage display (subtractive panning) 4 PMID:28881621 Gap junction alpha-5 protein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A phage library was first pre-cleared in an uncoated well, and then presented to the bait protein. One of the retrieved peptides (HSLRPEWRMPGP) showed a 58.3% homology with amino acids 5-to-16 of IκBα, a member of the protein complex inhibiting NFκB activation. 3253 NKLLATW(18) NKLCCTK(1) NKLCCAK(1) GQLLCTM(1) GQLLATK(1) LL 5 Phage display (common panning) 4 PMID:28853252 Anti-VP2 monoclonal antibody 3G11 Outer capsid protein VP2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The results of sandwich ELISA test shown that MAb 3G11 can react positively with BTV-8, and with other 23,no positive reaction was found in the virus species. After the fourth round of panning, obtained positive sequence clones KLLAT, the 3 amino acids LL and T of the sequence are consistent with the amino acid residue LLST sequence of BTV-8 VP2. 3254 SAPSSKN(2) HAIYPRH(2) AMLPYTF(1) ANSTPPI(1) ANTTPRH(1) ATLTHPP(1) EDRANRQ(1) EPLQLKM(1) GKPMPPM(1) GKVQAQS(1) GRPHSAL(1) HTAPNFA(1) IPTLPSS(1) LHRPYST(1) LPTPPYA(1) MTSQHPK(1) NQLPLHA(1) QILAFNS(1) SHGNWWR(1) SLLNRMP(1) SLYKWTl(1) SPTQPKS(1) STPIQQP(1) TLHSLPP(1) TSLSMPS(1) TTVMGNL(1) VAHQLSR(1) WTITKHP(1) 28 Phage display (in vivo) 3 PMID:28916756 Human pancreatic ductal carcinoma cell Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) To conduct the phage screen, 10 µL of PhD-7 phage display library (New England Biolabs, Ipswich, MA, USA) was combined with 190 µL of Dulbecco’s phosphate buffered saline (DPBS) and injected intravenously into mice harboring orthotopic xenograft PDAC tumors. Phage circulated for 8–10min before the animals were killed and cardiac perfused with 30mL of DPBS. Tumors were minced into 1–2mm segments and Dounce homogenized in a solution consisting of DPBS+ 1% TritonX, ethylene diamine tetraacetic acid, and protease inhibitors (Thermo Scientific, Waltham, MA, USA). To assess the specificity of a few of the selected clones for blood vessels in vivo, fluorescently labeled phage were injected into animals bearing orthotopic tumors. Specificity of the phage for tumor vessels was assessed by immunofluorescence. We used three clones with similar homology, PRH, in our analysis. The “PRH” pool (HAIYPRH and LHRPYST) bound 73.4% (4.1-fold over normal pancreas vessels) of platelet endothelial cell adhesionmolecule1-positive tumor vessels. The immunofluorescent images indicate that the fluorescein isothiocyanate (FITC)-phage remained bound to the luminal surface of the blood vessels even after vigorous perfusion. 3255 DLTFTVNPLSKA(8)[0.134, 1.616] WHWSWWHPNQLT(7)[0.550, 1.949] TSVSYINNRHNL(1)[0.122, 0.377] 3 Phage display (common panning) 3 PMID:28929019 Refolded apical merozoite antigen 1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding signals for each peptide were estimated using absorbance value at 405 nm towards BSA (control) and rPvAMA1, respectively. The first column of the affinity values is the absorbance value towards BSA, the second is towards rPvAMA1. 3256 YYWGPPV(2) HYIDFRW(2) WNEVKPI(1) IIIRNSF(1) NYTLTDI(1) AQNQPMA(1) SLLGQTP(1) GLFHNAQ(1) 8 Phage display (common panning) 3 PMID:28888997 Human prostate cancer cell line PC3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) PC-3 cells were seeded onto a well of a 96 well microplate (Nunc Maxisorb) for 24 hours to bring the culture to about 75% confluence. The cells were washed with PBS and then exposed to 1.0e9 phage particles from the parent Ph.D-7 library in 100µl PBS. After incubation, the cells were washed with PBS and then 100µl of 0.2M Glycine-HCl, pH. 2.2 were added to elute cell-bound phage. After 10 mins, 15µl of 1M Tris-HCl, pH 9.1 was added to neutralize the solution and the eluted phage were recovered. The cells were washed again with PBS and then 100µl 30mM Tris, pH. 8.0 was added. The cells were frozenthawed until they had ruptured as judged by light microscopy (three cycles). The ruptured cell mixture was collected and centrifuged at 2000rpm for 2 min; the supernatant containing the PC3 internalized phage was recovered. Phage were titrated, amplified and the exposure procedure was repeated twice more. 3257 TPARHIY(3) AYPSAHR(2) TPARHIY(1) THISKLI(1) NERALTL(1) YWHPHMV(1) LSNNNLR(1) 7 Phage display (subtractive panning) 3 PMID:28888997 Human prostate cancer cell line PC3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The first (termed L1) was the parent Ph.D-7 library. The second library (L2) was derived from L1 and comprised phage that did not bind normal mouse tissues. For this purpose, 1.0e11 pfu of L1 (10µl) in 100µl PBS were injected intravenously into a Balb/c mouse and phage were recovered from the blood 24hrs later using the phage isolation procedure described in the kit. Then three rounds of biopanning were performed against the PC-3 cells, which were seeded onto a well of a 96 well microplate (Nunc Maxisorb). Phage from a libraries displaying 7mer peptides were exposed to PC-3 cells and only internalized phage were recovered. The ability of these phage to internalize into other prostate cancer cells (LNCaP, DU-145) was validated. The displayed peptides of selected phage clones were synthesized and their specificity for target cells was validated in vitro and in vivo. One peptide (P12, LSNNNLR) which specifically targeted PC-3 tumors in vivo was incorporated into mono-drug (Chlorambucil, Combretastatin or Camptothecin) and dual-drug (Chlorambucil/Combretastatin or Chlorambucil/Camptothecin) PDCs and the cytotoxic efficacy of these conjugates for target cells was tested. Conjugation of P12 into dual-drug PDCs allowed discovery of new drug combinations with synergistic effects. 3258 CPRIWADSC(12)[4.84] CTYLNSAKC(10)[4.63] 2 Phage display (competitive panning) 3 PMID:28783899 Anti-imidaclothiz monoclonal antibody 1E7 Imidaclothiz Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The absorbance at 450 nm was measured. Data shown as affinity values were IC50 (ng/mL). The imidaclothiz (2 μg/mL in PBS) was added to elute the binding phage with shaking for 1 h. 3259 CLPPRMIYEC(25)[4.02] 1 Phage display (competitive panning) 3 PMID:28783899 Anti-imidaclothiz monoclonal antibody 1E7 Imidaclothiz Not determined. Not determined. CX8C M13 phage display library ELISA The absorbance at 450 nm was measured. Data shown as affinity values were IC50 (ng/mL). The imidaclothiz (2 μg/mL in PBS) was added to elute the binding phage with shaking for 1 h. 3260 TMHLPYCPTNIC(29)[2.97] SQPWCPPSICGD(8)[5.08] DYHDPSLPTLRK(1)[5.74] 3 Phage display (competitive panning) 3 PMID:28783899 Anti-imidaclothiz monoclonal antibody 1E7 Imidaclothiz Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance at 450 nm was measured. Data shown as affinity values were IC50 (ng/mL). The imidaclothiz (2 μg/mL in PBS) was added to elute the binding phage with shaking for 1 h. 3261 DRWVARDPASIF(22/60) FPSSPGVSRAIN(3/60) GLHTSATNLYLH(2/60) GVAMLQSGYASR(2/60)[0.36] VTVELQHLSPAK(2/60)[0.45] FEHSLYKEMTHL(1/60)[1.79] NYPATNTHRYTP(1/60) TPQSFWQKGSLV(1/60) AFDTFNLYMDEL(1/60) VPGLVFSKLTTG(1/60) VPAQSGQGIEHE(1/60) SPDLSTRPTLGY(1/60) IPAPLLVPNLWH(1/60) SSDSVVKQAILT(1/60) HMNTHDSLFPVG(1/60) QPNNILPAFGNY(1/60) APTLYWNVPTHV(1/60) LFAAASSNHIHR(1/60) SVLPLLCCDSTP(1/60) VERSFLNERVTP(1/60) DVTRLLSTHMWI(1/60) GIQNGLAFQGLQ(1/60) QLQLNNQVALMY(1/60) 23 Phage display (subtractive panning) 5 PMID:28823053 Macrophage-stimulating protein receptor, MSP receptor Hepatocyte growth factor-like protein 4QT8, Not determined. Ph.D.-12 phage display library (X12) ELISA A subtractive biopanning was first performed by adding a peptide-displaying phage library to pre-cold NIH3T3 cells. Subsequently, the supernatant was transferred onto pre-cold NIH3T3-RON cells and incubated. Totally, five rounds of biopanning were carried out using NIH3T3-RON cell line. Then,the recovered phage from fifth round were used for subtractive biopanning using EBC-1-Dox+ (RON negative) in order to eliminate possible non-RON binding phage particles. Binding modes and affinities were also predicted by docking and molecular dynamics (MD) simulation. The results of ELISA experiment showed that P6 peptide (FEHSLYKEMTHL) displaying phage has higher affinity for RON compared to others and its binding site is located out of ligand binding site. Docking and MD simulation results also indicated higher affinity of P6 to RON as well as its exosite-binding feature. Taken together,our data suggest a capacity for P6 peptide to be utilized as RON binding agent,and hence be used for various purposes,including design of drug delivery systems for transferring cytotoxic agents to RON-positive cancer cells, interfering with RON signaling, peptidomimetics design, and diagnostic imaging. 3262 DRWVARDPASIF(39/60) FPSSPGVSRAIN(20/60) 2 Phage display (subtractive panning) 5 PMID:28823053 Macrophage-stimulating protein receptor, MSP receptor Hepatocyte growth factor-like protein 4QT8, Not determined. Ph.D.-12 phage display library (X12) A subtractive biopanning was first performed by adding a peptide-displaying phage library to pre-cold MDCK cells. Subsequently, the supernatant was transferred onto pre-cold MDCK-RON cells and incubated. Totally, five rounds of biopanning were carried out using MDCK-RON cell line. Then,the recovered phage from fifth round were used for subtractive biopanning using EBC-1-Dox+ (RON negative) in order to eliminate possible non-RON binding phage particles. 3263 NCWSSLRGICENLG[21.7] SCSWNVERIRGCSV[18.8] CWNLKRIGSQGC[30.7] CWNLKRIGSQGC[26.4] GCVKYRGSVSCQESKT[18.0] ACLNHFVSGNMIRC[26.1] CPFYDWRCSDF[25.0] NVRCYRDVPSCS[25.9] EDVVCYGRVDYMPLCV[25.2] DCNLWGDDGKYRLCFG[23.6] CVKVGLSWIGDCNS[23.9] CMEAWQCSLA[24.6] QCYPWCRL[25.1] CWTYLRGGCSVT[22.1] GDCGYHYMCSMT[25.0] NMCYLRGKMDCNIV[30.9] VCSDHYIGGKEVRC[27.4] CCSYGYEKCCNG[28.6] SPCRVGYTPC[27.7] NCARLYSHQRDYSQVC[24.4] CLDRSGGCYTREGYLS[29.0] CYPLCRVG[16.2] KCDSHYIRGVVACH[30.2] GCGYRYMCDMINGL[32.8] GDCYPHCRLV[28.9] DCLRYRVGSSCS[29.6] RCESKGWANTCVSE[20.7] CYPLCRVL[31.1] RCGLAWCVVLVDQ[20.8] 29 Phage display (common panning) 4 PMID:28825773 Proprotein convertase subtilisin/kexin type 9, PCSK9 Low-density lipoprotein receptor 2W2M,2W2N,2W2O,2W2P,2W2Q,3BPS,3GCW,3GCX,3M0C,3P5B,3P5C, Not determined. TAFTSWEEYLDWVGSG fusion M13 phage display library (TAFTSWEEYLDWVGSGX(8,10,12,14,16)) ELISA The s/n ratio is the signal:noise ratio, wherein “signal” and "noise" are the spot phage ELISA signals of phage binding to NeutrAvidin-captured PCSK9 and to NeutrAvidin, respectively. The s/n ratios were shown. 3264 KLWNLGRV[28.2] VLWNHSRI[24.9] GPWNLNRV[27.6] PLLWNLQKVH[23.5] KLWNLTRISS[28.1] VVLWNHSRIL[26.8] VPWNLARL[29.3] VPWNLAKI[22.5] GHPLWNLSRI[25.7] NELWNINRLRSS[12.0] SIPWNLERITPPR[28.9] VLWNHSRI[29.7] MPWNLARIER[27.8] ALWNMRRVESVAFR[23.9] GEYLWNLKRLES[28.7] TRYADRGVMVYSLS[23.0] MVYVDRGVRVFT[28.0] EHQFYSYRGVDVYR[22.3] KNNYAYLINMPRAPGI[24.7] RGGYELNIPR[26.1] NPTYWLTRIS[23.1] PHSYWLDRVQ[25.3] GVLGVGGMWIA[12.8] SYRGAWSVFGGSTD[19.5] PRTYLRGLVD[25.6] QTTYLRVSSS[27.0] QISYLRNA[22.9] RGEVYDKGGVIVHL[30.5] GREMRSVIQM[28.2] QELPNHRRLS[19.1] SETGLGTYYG[28.1] SSTWHFVGGVRV[24.4] TRYADRGVMVYSLS[30.3] KMTHMKAD[22.4] PTSGLTPS[27.5] QVTSLPQA[27.9] RGGYELNIPR[29.8] W-N-L-x-R-I 37 Phage display (common panning) 4 PMID:28825773 PCSK9ΔP' Not determined. Not determined. Not determined. WNLVRIGLLR fusion peptide library (X(2-5)(G/S)WNLVRIGLLR) ELISA The s/n ratio is the signal:noise ratio, wherein “signal” and "noise" are the spot phage ELISA signals of phage binding to NeutrAvidin-captured PCSK9 and to NeutrAvidin, respectively. The s/n ratios were shown. One cluster contained a motif, WNLxRI , homologous to the sequence of the native P' helix. 3265 HLWGWLYAPSFQ(15)[0.95] YTFHFDIFQPHF(1)[25.66] 2 Phage display (subtractive panning) 6 PMID:28986753 Galectin-1, Gal-1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA To determine the apparent dissociation constant (K*d) of the selected clones, ten serial dilutions of phages (generally ranging from 5e12 to 1.0e7 phages/100 µL) were incubated with gal-1 or control proteins in the same conditions as for the first clones' screening. The optical density (OD) was measured (405 nm, differential filter: 630 nm) using a microplate reader (StatFax-2100, Awareness Technology, Fisher Bioblock Scientific, Tournai, Belgium). The affinity constants (Ka = 1/K*d) against gal-1 or control proteins were determined for one representative clone per peptide sequence. The phage clone has a better specific affinity when the affinity ratio is high. Coll = collagen. Affinity ratio = KaGal-1/((KaHSA + KaColl)/2). Data shown are affinity ratios. Based on this parameter, the best peptide clone is YTFHFDIFQPHF (P2) A linear 12-mer random peptide phage display library (PhD-12, New England Biolabs Inc. Bioké, Leiden, The Netherlands) was screened against human gal-1 (Peprotech, London, UK) for six rounds, which was immobilized on a 96-well ELISA plate at a concentration of 100 µg/mL. The preselection step was performed by incubating the phage library with bovine serum albumin (BSA, 1 µg/mL, Sigma-Aldrich, Diegem, Belgium). The time for the preselection step is modified along the rounds (80 – 100 – 120 – 140 – 160 –180 min). Peptide P1 (HLWGWLYAPSFQ) was selected according to its affinity toward gal-1. Its binding has been revealed by immunohistochemistry on human thyroid cancer biopsies, and it was co-localized with gal-1 by immunofluorescence on TPC-1 cell line. Peptide P1 induced a decrease in TPC-1 cells' adhesion to gal-1 immobilized on culture plates. After coupling to USPIO, the peptide preserved its affinity toward gal-1. Its specific binding has been corroborated by co-localization with gal-1 expressed by TPC-1 cells and by its ability to compete with anti-gal-1 antibody. The peptide and its USPIO derivatives produce no toxicity in HepaRG cells as determined by MTT assay. The vectorized contrast agent is a potential imaging probe for thyroid cancer diagnosis. Moreover, the gal-1-targeted peptide prevent cancer cell adhesion by interacting with the carbohydrate-recognition domain of gal-1. 3266 HLWGWLYAPSF(15)[0.95] HYSWSWIAYSPG(2)[1.14] VHWDFRQWWQPS(2)[1.47] YSWHIDIVAPRN(2)[Infinite ratio] YHHSGPYAGPMW(1)[2.50] SVYVEISWVRTM(1)[1.17] DWSSWVYRDPQT(1)[2.36] 7 Phage display (subtractive panning) 3 PMID:28976753 Galectin-1, Gal-1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Peptide P7 (YSWHIDIVAPRN) was selected according to its affinity toward gal-1. Its binding has been revealed by immunohistochemistry on human thyroid cancer biopsies, and it was co-localized with gal-1 by immunofluorescence on TPC-1 cell line. Peptide P7 induced a decrease in TPC-1 cells' adhesion to gal-1 immobilized on culture plates. After coupling to USPIO, the peptide preserved its affinity toward gal-1. Its specific binding has been corroborated by co-localization with gal-1 expressed by TPC-1 cells and by its ability to compete with anti-gal-1 antibody. The peptide and its USPIO derivatives produce no toxicity in HepaRG cells as determined by MTT assay. The vectorized contrast agent is a potential imaging probe for thyroid cancer diagnosis. Moreover, the gal-1-targeted peptide prevent cancer cell adhesion by interacting with the carbohydrate-recognition domain of gal-1. A linear 12-mer random peptide phage display library (PhD-12, New England Biolabs Inc. Bioké, Leiden, The Netherlands) was screened against human gal-1 (Creative Biomart, New York, USA) for three rounds, which was immobilized on a 96-well ELISA plate at a concentration of 100 µg/mL. The preselection step was performed by incubating the phage library with a mix of human serum albumin (HSA) and collagen (both at 100 µg/mL; Sigma-Aldrich). The time for the preselection step is modified along the rounds (80 – 100 – 120 min). Peptide P7 (YSWHIDIVAPRN) was selected according to its affinity toward gal-1. Its binding has been revealed by immunohistochemistry on human thyroid cancer biopsies, and it was co-localized with gal-1 by immunofluorescence on TPC-1 cell line. Peptide P7 induced a decrease in TPC-1 cells' adhesion to gal-1 immobilized on culture plates. After coupling to USPIO, the peptide preserved its affinity toward gal-1. Its specific binding has been corroborated by co-localization with gal-1 expressed by TPC-1 cells and by its ability to compete with anti-gal-1 antibody. The peptide and its USPIO derivatives produce no toxicity in HepaRG cells as determined by MTT assay. The vectorized contrast agent is a potential imaging probe for thyroid cancer diagnosis. Moreover, the gal-1-targeted peptide prevent cancer cell adhesion by interacting with the carbohydrate-recognition domain of gal-1. 3267 CIGNSNTLC(38.4%)[6.29] CTVRTSADC(23.2%)[15.4] CSNNGNALC(15.4%)[47.3] CISNGNQPC(15.4%)[64.2] CRVNTAALC(7.6%)[77.7] 5 Phage display (common panning) 3 PMID:29029605 Immunoglobulins (Igs) of murine 5T33MM cells Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The specific binding of peptides to the 5T33MM sIgG was analyzed by ELISA. Streptavidin coated 96 well plates were washed extensively and supplemented with biotin-conjugated peptides by 1 h-incubation at 37 °C; after washing and blocking with blocking solution (1X PBS, 0.05% Tween-20, 5% milk), aliquots of 5T33MM sIgG were added overnight at 4 °C. Wells were extensively washed and coated with the anti-mouse IgG (Fc-specific) alkaline phosphatase-conjugated [Sigma Aldrich] for 1 h at 37 °C, incubated with the alkaline phosphatase substrate, and analyzed by an ELISA reader at 405 nm [Labsystems multiscan MS]. KD values (nM) of identified synthetic peptides are shown. The screening of phage displayed library was performed using the bait 5T33MM Igs. Briefly, the streptavidin-conjugated beads were coated with 5T33MM Igs and incubated with 1.0e11 phages overnight at 4 °C. 5T33MM Igs-interacting phages were eluted with 0.2 M glycine-HCl (pH 2.2, 1 mg/mL BSA) followed by the addition of neutralizing solution (1 M Tris-HCl pH 9.1). Ultimately, 3 cycles of panning were performed. Plaques of lysis from isolated phages were transferred to nitrocellulose filters, and membranes were blocked with PBS 1X, 0.1% NP-40; 5% milk; 0.02% NaN3 and then incubated for 2 h at RT with 100 μg of purified 5T33MM Igs. After washing, membranes were hybridized with an alkaline phosphatase-conjugated anti-mouse IgG (dilution 1:5000) and then washed 6 times. Immunoreactive phage clones were detected by BCIP/NBT premixed substrate. Based on the highest affinity binding to 5T33MM Igs, the insert amino acid sequence of phage clone 5 (CIGNSNTLC) was used for large-scale synthesis of the p5 peptide. By FACS, the FITC-conjugated P5 detected the MM-released exosomes in the serum of 5T33MM-engrafted mice, levels of which are correlated with tumor progression at an earlier time point compared to serum paraprotein. 3268 DWSSWVYRDPQT(3)[37933.89] LPKTVSSDMSLN(1)[NT] 2 Phage display (subtractive panning) 4 PMID:29065111 Human Colon cancer cell line COLO320HSR Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Isothermal titration calorimetry (ITC) COLO320HSR was grown on coverslips overnight and fixed with 4% paraformaldehyde at room temperature for 30 min. Samples were blocked in 3% BSA (w/v) inPBS for 30 min at room temperature. Cells were then incubated with 10 mM FITC-labelled peptides for 15 min at room temperature and washed with PBS for three times. This was followed by nuclear staining with 4,6-diamidino-2-phenylindole (DAPI). A laser scanning confocal microscope was used to visualise the slides. The cumulative values of the integrated optical density (IOD) of green fluorescence (FITC) in each photo were analysed using the Image-Pro Plus 6.0 software. The IOD value was reproduced from Figure 3D in the literature and shown. Four rounds of reiterative biopanning were performed against a human colon cancer cell line (COLO320HSR). Then, the selected phages were applied to normal human intestinal epithelial cell line NCM460 in the same way, for subtractive screening. The peptides that specifically binds to human colon cancer cell line COLO320HSR but not to a normal human intestinal epithelial cell line NCM460 were selected finally. A peptide termed as CBP-DWS with the sequence of DWSSWVYRDPQT, which was demonstrated to be capable of binding to a panel of human colon cancer cell lines and tissues, was identified; it had virtually no binding to normal human intestinal epithelial cell line NCM460 and normal surrounding colon tissues. Bioinformatics analyses suggest that CBP-DWS targets human Glypican-3, which may be involved in important cellular functions in multiple cancer types. 3269 TLLVIRGLPGAC(38)[1.54] NLLTMARPSWLH(20)[0.99] ANPKVRVGHAAS(20)[0.80] VKKRTLCAAACC(6)[0.28] VKVTTASCCSMW(4)[0.31] HLMDHPIRSNNQ(1)[0.30] TTRSSAFLIGGP(1)[0.29] 7 Phage display (common panning) 5 PMID:23031494 Gold Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA To verify specificity of selected phages, two different and independent assays were conduced. Wild-type (Wt-M13) phages with no displayed peptide on their surface were used as negative control. Results were analyzed by an ELISA reader (LabSystem Multiscan MS) at 405 nm. The intensity of absorbance at 405 nm from selected phages adsorbed on gold surfaces was reproduced from Figure S6 in the literature. The intensity of absorbance of the Wt-M13 phage was 0.27. In the phage context, we demonstrated the adhesive motif was capable to adsorb on gold in a preferential way with a morphological and viscoelastic signature of the adsorbed layer as evidenced by QCM-D and AFM investigations. Out of the phage context, the linear dodecapeptide (TLLVIRGLPGAC) is reproducibly found to adhere to the gold surface, and by quantitative SPR measurements, high affinity constants (Keq ~ 1.0e6 M-1, binding energy ~ -8kcal/mol) were determined. We proved that the interactions occurring at gold interface were mainly hydrophobic as a consequence of high frequency of hydrophobic residues in the peptide sequence. Moreover, by CD, molecular dynamics and steered molecular dynamics, we demonstrated that the molecular flexibility only played a minor role in the peptide adsorption. 3270 CKKNSPTLC(15/24) CIGNSNTLC(7/24) CAKATCPAC(2/24) 3 Phage display (subtractive panning) 4 PMID:28029647 G-protein coupled receptor 55 Lysophosphatidylinositol, LPI Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Briefly, phages of approximately 1.0e10 plaque-forming units (PFU) were pre-incubated for 1 h with 5.0e5 wild-type HEK293 cells. The supernatant containing unbound phage particles was subsequently transferred to 35-mm Petri dishes to be incubated with 5 ×105 ev-HEK293 cells for an additional 2 h, to further eliminate phage particles with a specific binding. The ultimate pre-cleaned supernatant was incubated with GPR55-HEK293 cells for 2 h, and unbound phage particles were removed by washing with 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1% Tween-20. The extracellular bound phages were eluted with 100 mM glycine-HCl (pH 2.2) for 10 min, and neutralized with 1 M Tris-HCl (pH 9.0). The plasma-membrane-bound phages were collected. Sequencing of selected phagotopes dictated the primary structure for the synthetic FITC-labeled peptide P1 (CKKNSPTLC), which was analyzed for binding specificity to immunoprecipitated HA-GPR55, and to endogenously expressed the lysophosphatidylinositol receptor GPR55, using cells interfered or not for GPR55, as well as for co-localization imaging with HA-GPR55 at the membrane level. The peptide P1 stimulated GPR55 endocytosis and inhibited GPR55-dependent proliferation of EHEB and DeFew cells, two human B-lymphoblastoid cell lines. Our data support the potential therapeutic application of peptide ligands of GPR55 for targeting and inhibiting growth of neoplastic cells, which overexpress GPR55 and are dependent on GPR55 signaling for their proliferation. 3271 CKKNSPTLC(14/18) CIGNSNTLC(4/18) 2 Phage display (subtractive panning) 4 PMID:28029647 G-protein coupled receptor 55 Lysophosphatidylinositol, LPI Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Briefly, phages of approximately 1.0e10 plaque-forming units (PFU) were pre-incubated for 1 h with 5.0e5 wild-type HEK293 cells. The supernatant containing unbound phage particles was subsequently transferred to 35-mm Petri dishes to be incubated with 5 ×105 ev-HEK293 cells for an additional 2 h, to further eliminate phage particles with a specific binding. The ultimate pre-cleaned supernatant was incubated with GPR55-HEK293 cells for 2 h, and unbound phage particles were removed by washing with 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1% Tween-20. The extracellular bound phages were eluted with 100 mM glycine-HCl (pH 2.2) for 10 min, and neutralized with 1 M Tris-HCl (pH 9.0). An additional step of cryogenic lysis of GPR55-HEK293 cells allowed the recovery of the intracellular bound phages. The internalized phages were collected. Sequencing of selected phagotopes dictated the primary structure for the synthetic FITC-labeled peptide P1 (CKKNSPTLC), which was analyzed for binding specificity to immunoprecipitated HA-GPR55, and to endogenously expressed the lysophosphatidylinositol receptor GPR55, using cells interfered or not for GPR55, as well as for co-localization imaging with HA-GPR55 at the membrane level. The peptide P1 stimulated GPR55 endocytosis and inhibited GPR55-dependent proliferation of EHEB and DeFew cells, two human B-lymphoblastoid cell lines. Our data support the potential therapeutic application of peptide ligands of GPR55 for targeting and inhibiting growth of neoplastic cells, which overexpress GPR55 and are dependent on GPR55 signaling for their proliferation. 3272 LMSTCTWLDECFRPQ(6)[1.025] PTPPCHRGDECQPLW(5)[0.703] SRPHCYPMDDCHPLW(4)[1.137] WSGMCESQWCKDFA(4)[1.039] TPHTCKLLDECVPLW(3)[2.925] TPTVCTWLDECPPWS(1)[1.052] HTPWCSIADPCLWEL(1)[1.597] YHEPCLWATACPTTP(1)[0.464] MQDMCNDDSCPLWS (1)[0.481] CTHPCEPPPLWPIAP(1)[2.082] PGPQCPYPAELWCTQ(1)[0.336] 11 Phage display (competitive panning) 2 PMID:24356816 Anti-CD43 monoclonal antibody UN1 Leukosialin Not determined. Not determined. f88-Cys5 phage display library (X4CX5CX4) ELISA Optical density (OD) at 405 nm (OD405) and 620 nm (OD620) were measured by an ELISA reader (Tecan), and values were expressed and shown as difference between OD405 and OD620. The UN1 mAb (10 μg) was linked to streptavidin-conjugated magnetic beads (Promega), which had been coated with 10 μg of goat anti-mouse IgG (Fc-specific) biotin-conjugated Ab (Sigma-Aldrich) in 200 μL of beads suspension. A total of 3.0e10 transducing units of phages from the library were added and incubated for 16 hours at 4 °C. After extensive washing, bound phages were eluted with 0.1 mol/L HCl/glycine buffer, pH 2.2, 1 mg/mL bovine serum albumin (BSA), and neutralized with 1 mol/L Tris, pH9.1. Eluted phages were amplified by infection of K91BK bacteria, and purified from plaques for a second round of affinity selection. Phage colonies were transferred according to an ordered grid on a lawn of K91BK cells on Luria-Bertani (LB) agar plates supplemented with 1 mmol/L isopropyl b-D-1-thiogalactopyranoside. Nitrocellulose filters were layered onto these plates and incubated overnight at 37 °C. Filters were blocked for 2 hours with blocking buffer(1 × PBS, 5% non-fat dry milk, 0.1% NP40, 0.01% NaN3) at room temperature and incubated O/N at 4 °C with the UN1 mAb (1 μg/mL) in blocking buffer. Then, filters were washed with washing buffer (1 × PBS, 0.1% NP40) and incubated with alkaline phosphatase-conjugated anti-mouse IgG (Fc specific) secondary antibody (SigmaAldrich) at the dilution of 1:5000 for 4 hours at 4 °C. After extensive washing, filters were incubated in developing solution (1-Step NBT/BCIP, Pierce, Thermo Scientific). Collected phage supernatants from positive clones were further analyzed by ELISA. By screening a phage-displayed random peptide library, we identified the phagotope 2/165 with the sequence of TPHTCKLLDECVPLW as a mimotope of the UN1 antigen, as it harbored a peptide sequence that was specifically recognized by the UN1 mAb and inhibited the binding of the UN1 mAb to UN1-positive tumor cells. On the basis of sequence homology with the extracellular region of CD43 (amino acids 64 to 83), the 2/165 peptide sequence was likely mimicking the protein core of the UN1/CD43 epitope. When used as vaccine in mice, the 2/165 phagotope raised antibodies against the UN1/CD43 antigen, indicating that the 2/165 phagotope mimicked the UN1 antigen structure, and could represent a novel immunogen for cancer immunotherapy. 3273 CIIGSYVC CNLSVPAC CLSPIGEC CLGKGSVC CGRDSKQC CAAAPIRC CAGGGAYC CMRGKGLC CASGSENC CVPMRLQC CGRAKVRC CLPISSSC CMARQARC CRECGERC CAGKSSNC CIRREKRC CNIRRQGC CPQPRPLC CLANLQTC CAVVFSQC CSIRRESC CWRSVEVC CADVLRPC CEGESASC 24 Phage display (subtractive panning) 3 PMID:11689892 Human endothelial cells stimulated with vascular endothelial growth factor (VEGF) Not determined. Not determined. Not determined. CX6C phage display library A method termed biopanning and rapid analysis of selective interactive ligands (BRASIL) was used. BRASIL allows separation of phage-cell complexes from the remaining unbound phage; this is accomplished by a differential centrifugation that drives the cells from a hydrophilic environment into a non-miscible organic phase. Because the organic phase is hydrophobic, it excludes water-soluble materials surrounding cell surfaces. Bound phage are recovered from the cell pellet whereas the unbound phage remain soluble in the upper aqueous phase, eliminating the need for repeated washes. A two-step panning strategy to isolate phage that bind to angiogenic ECs was used. First, to decrease non-specific binding, we pre-cleared the phage library on starved ECs (before panning on the same cell line stimulated with recombinant VEGF165); starved human umbilical vein ECs (HUVECs) were incubated with the phage library and centrifuged through the organic phase. Second, the unbound phage pool left in the aqueous phase was transferred to a fresh tube and incubated with VEGF165-stimulated HUVECs. After centrifugation through the organic phase, phage bound to the VEGF165-stimulated HUVEC pellet were recovered by bacterial infection, amplified and subjected to two more rounds of selection. The motif PQPRPL is a novel chimeric ligand mimic that binds specifically to VEGF receptor-1 and to neuropilin-1. 3274 PFTVSVPFVWNFTAD(28/60)[0.59 ± 0.01] WAGPRVSSVYYAGAR(6/60)[0.72 ± 0.01] 2 Phage display (common panning) 3 PMID:29156833 Anti-leptospira monoclonal antibody P17 (8C1E11) Not determined. Not determined. Not determined. f5-15mer phage display library (X15) ELISA Eleven phage clones with the highest selection frequency and unique sequences were chosen for analysis of their ability to bind the v6 region peptide using phage ELISA. The optical densities were measured with a μQuant Universal Microplate Spectrophotometer (Bio-Tek Instruments, Winooski, VT) at 490 nm. The data shown were reproduced from Figure 1C. Libraries were selected against a v6 region peptide derivative, KEQWFGNRWHEGYR, which encompassed a 14 amino acid sequence of v6 domain of human CD44v6 protein. After three cycles of affinity selection, sixty individual phage clones from the elution pool of the fUSE5 library were randomly chosen for DNA sequencing. A 15 amino acid peptide (PFTVSVPFVWNFTAD, PFT) was identified by affinity selection against a peptide derived from the v6 region of CD44 (CD44v6). Synthesized PFT exhibited specific binding to CD44v6 with an equilibrium dissociation constant (Kd) of 743.4 nM. PFT also bound CD44v6 highly expressed on human PCa cell lines. Further, an aggressive form of PCa cells (v6A3) was isolated and tagged by a novel CSC reporter vector. The v6A3 cells had a CSC-like phenotype including enriched CD44v6 expression, enhanced clonogenicity, resistance to chemotherapeutics, and generation of heterogeneous offspring. PFT exhibited preferential binding to v6A3 cells compared to parental cells. Immunohistofluorescence studies with human PCa tissue microarrays (TMA) indicated that PFT was highly accurate in detecting CD44v6 positive aggressive PCa cells, and staining positivity was significantly higher in late stage, metastatic and higher-grade samples. Taken together, this study provides for the first time phage display selected peptides that target CD44v6 overexpressed on PCa cells. Peptide PFT may be explored as an aid in the diagnosis and therapy of advanced PCa disease. 3275 CSAYDRPLC(12/60)[0.74 ± 0.01] CWSNKGYDC(2/60)[0.79 ± 0.01] 2 Phage display (common panning) 3 PMID:29156833 Peptide KEQWFGNRWHEGYR Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Eleven phage clones with the highest selection frequency and unique sequences were chosen for analysis of their ability to bind the v6 region peptide using phage ELISA. The optical densities were measured with a μQuant Universal Microplate Spectrophotometer (Bio-Tek Instruments, Winooski, VT) at 490 nm. The data shown were reproduced from Figure 1C. Libraries were selected against a v6 region peptide derivative, KEQWFGNRWHEGYR, which encompassed a 14 amino acid sequence of v6 domain of human CD44v6 protein. After three cycles of affinity selection, sixty individual phage clones from the elution pool of the M13C7C library were randomly chosen for DNA sequencing. 3276 YLWPDYPAPSLP(12)[6.14 ± 1.95] WVRVRSMLPVAP(11)[8.59 ± 1.81] WLHQECARMGAL(11)[23.19 ± 4.87] TCSRLAPWACVT(11)[8.61 ± 3.09] VFRGSPGYVEYV(10)[30.40 ± 2.86] PIPPLRWFEERV(10)[10.62 ± 2.00] WWTTKAFLSGPW(9)[31.12 ± 2.20] WSFLVPPSFRPI(9)[29.40 ± 5.94] GWYWTADLMDKM(9)[8.68 ± 3.57] TFHLMRPLMLPV(8)[17.57 ± 3.05] VPLRHSSLVWLV(7)[7.07 ± 2.65] WPDWTPRLPVPY(6)[37.74 ± 4.16] VRVAPSVMAPVL(6)[11.06 ± 4.93] LRPVVASKSWVI(6)[8.52 ± 5.38] YPAVPPRVVAAI(5)[23.98 ± 12.71] GFTWKGYCSELV(5)[12.96 ± 5.97] YPPVYRAEKTLA(3)[11.65 ± 0.33] WRGSLSYLKGPL(3)[13.32 ± 8.83] WRCIGSWVSASY(3)[19.03 ± 4.04] WLYDRVHSMWVL(3)[31.25 ± 2.87] WKSCGVWAGCPM(3)[10.89 ± 3.01] SVMFSWPAPMIP(3)[32.75 ± 1.83] RWYKPLPSLLLW(3)[28.92 ± 4.52] RCIYPAYTGCLF(3)[3.48 ± 0.41] MPARPTLPTGLI(3)[27.72 ± 3.83] HGWHCVYGESYV(3)[4.32 ± 1.08] YRTRALPSLVVR(2)[7.66 ± 2.95] WVYRGNLGIWVL(2)[35.54 ± 0.16] WSTGSYWGSLLW(2)[7.35 ± 1.26] WRGGPAYLKYIS(2)[5.72 ± 0.41] WRDSSGYVMGPW(2)[33.72 ± 1.11] VLLTHSQWSKLY(2)[14.49 ± 9.79] VFWRTALAIPGG(2)[23.43 ± 2.06] STRSQLPWHWSQ(2)[13.91 ± 0.34] LSRYGYAYEAYV(2)[4.88 ± 1.88] GWRWEYSYLVGP(2)[33.31 ± 1.02] GVVRGDAYWFYV(2)[5.28 ± 2.12] GRPAIRIPDIIG(2)[24.17 ± 1.58] GPPVSVKSWVSQ(2)[8.73 ± 5.69] GPIVPVPSWLGI(2)[40.11 ± 0.85] GLLDHRMWNELY(2)[24.30 ± 0.58] AAPVVPSVPLNW(2)[4.73 ± 0.40] YLVGTMAPWWPA(1)[6.18] YFWHDLTGTWSV(1)[4.04] YDPRGYEIPSWL(1)[41.05] YCWWVSFTDCPL(1)[14.10] WYREPMPTPVVM(1)[10.76] WWWPGYDAYMSV(1)[37.69] WWWDSWNGFELL(1)[4.58] WWMESTFLEGPS(1)[41.43] WWFLDSYVNGPL(1)[28.46] WRALPNPGVSYV(1)[24.50] WPDWTPTLQVPW(1)[36.99] WLTYPAPPTLPF(1)[4.32] WLHLHPPIWAPL(1)[9.72] WLFSETDQVWYL(1)[18.43] WGWWLLDDVITS(1)[4.39] WGRYLACYHHLD(1)[9.71] WGPVVAPRIESA(1)[6.43] WEFVRANLPVWG(1)[12.05] WCLLVPHSTCAV(1)[6.48] WARRQLPALPVV(1)[35.70] VSLAGRVAAVLP(1)[11.53] VRMLPTPPNYFD(1)[4.44] VRAPAVIPRVTY(1)[4.93] VPNFLPPAVKNL(1)[26.70] VPKVPSAPAYYL(1)[4.49] VPAVNRLAKFVE(1)[26.14] VLPRFQAPSPTV(1)[20.80] VFDHVRGQWVAI(1)[26.41] VAAKELWWWDVV(1)[20.99] TPWWRAAIPSSL(1)[14.46] TGPLTAPRSVLY(1)[11.41] TAPCLPTLPWWQ(1)[20.62] SVPVLPYRAVTL(1)[4.14] SRPAFPMPGIVS(1)[3.06] SRFSFWLPVPAG(1)[24.99] SLPGWAPARFVS(1)[5.87] SLAVKWAAYPGL(1)[3.56] SGRPVPPVRWVM(1)[9.82] SGPTVPPVMWMM(1)[10.59] SEVPLFRPILRV(1)[30.35] SAWMRNLPLVPQ(1)[30.03] RWWWGGVPVNLI(1)[8.19] RWSIWHTGYLLV(1)[13.89] RWSHGYLAYMLD(1)[8.26] RWGWSFDPWLSM(1)[3.25] RWGWCFDRSLLD(1)[3.80] RVVKDWARLAIP(1)[3.60] RVVAPYRIPDWL(1)[35.37] RVPPRIVAPLYG(1)[13.44] RRPVVASNSWVI(1)[6.59] RQTYSSYIAMAM(1)[3.87] RQKMPIPVGNLV(1)[20.55] RPPVVASNSWVI(1)[15.42] RPDKPVRFVVSG(1)[10.97] RPAIPSRPIQFQ(1)[17.16] RLQRPLPALPCK(1)[31.17] RLPLRPPLPHTS(1)[5.48] RIVYGGGRWFLM(1)[6.01] RIRPAVALPYLL(1)[3.06] RIPNFIPMGPMM(1)[24.72] RGSVGYLTYLEH(1)[8.08] RETFSTYWLRTH(1)[5.97] RAPMPVGQSWMV(1)[18.41] RACFFIQGAVRC(1)[8.67] QMLPSFSPAEVI(1)[26.64] QGCCFVRSLPLM(1)[22.51] PWSRALPPLPGD(1)[3.28] PWATWGILSPEL(1)[29.94] PRYRLLPLYPYL(1)[32.37] PRRPQLPVVAVS(1)[7.86] PPLTAPRFWAEI(1)[28.56] PPAIPSGFCIGF(1)[18.13] PLVDRHLKYMGV(1)[29.21] PKYSAGLPAVPQ(1)[5.35] PAVGSSYACPGL(1)[23.73] NRPKAQLPTCYL(1)[14.57] NLQLPKWSPAAI(1)[22.07] NIMFGRLAAVLP(1)[23.10] NIGRARFMVPFL(1)[3.49] NGWALFKQTYMV(1)[28.55] MPHRGPLSLPTA(1)[3.49] MGNRARMSIPFV(1)[14.14] MFPTFIPPLPGK(1)[6.12] LVPALPEYSWNW(1)[15.87] LSYVVRPLPELP(1)[11.41] LSQRARLELPWS(1)[12.19] LRYGSAYLRYVS(1)[10.61] LRTGEAYLRYVD(1)[10.12] LRGAYPFHSWMV(1)[6.61] LPLPPIPPSYIA(1)[14.85] LPFHSRANPHYL(1)[8.36] LIPVRPVLPMAM(1)[17.35] LIAVKPLRTAFI(1)[5.34] LFVVKPLRLSVL(1)[10.22] LAYRSRAFLALP(1)[8.93] LAVPTRPSLPWT(1)[27.27] LALRSLPLVGAQ(1)[3.82] KRYSALPRVTDT(1)[30.90] ISYIGPEVASRA(1)[10.65] IIWSPVSSWWDL(1)[3.06] IGRAQFPLPIIS(1)[24.68] ICFGSGVGWYEC(1)[4.88] HRLFRLPLPVPV(1)[20.67] HPAVPPRVVAAI(1)[32.89] GYWKWSPVLDIV(1)[4.54] GYFPVIPTSVAN(1)[14.78] GWYYFQGMMGTS(1)[3.04] GWYCNYQTSTVF(1)[4.41] GWYCEYVYQARQ(1)[3.59] GWWWVESPLKVI(1)[3.06] GWWFLEPPSGAP(1)[5.67] GWRVTAAYLTYG(1)[13.87] GWRGSYAFLQGP(1)[31.03] GVKWYRRAPLLT(1)[17.19] GVAPGLPLYGPW(1)[10.42] GRYPLVNWVLFP(1)[22.87] GRTYPA*SGCWL(1)[3.57] GRKPPLLMAPIY(1)[8.06] GPWFLDGYGVWW(1)[6.81] GPSVSERTFFYL(1)[3.63] GPSVPSLPLDFG(1)[29.98] GPPVWPRMPGLV(1)[4.44] GPPVNYATKYEV(1)[15.00] GPMVWPRQVVYV(1)[14.34] GPAVPRLTVCYT(1)[10.43] GNAPATMQRYFV(1)[4.23] GLKRYLPLLTWT(1)[28.74] GIRGRSFWLPVP(1)[27.42] GGACQYVQHWSC(1)[5.30] GFSHPEWPAPML(1)[10.63] GAPALPMRSWWL(1)[18.91] FPSWWGDGPVGV(1)[7.16] ESYIPAVGAVAF(1)[5.72] DIRNRDLPPLPL(1)[19.07] DIPQFWFRGVFY(1)[6.15] CYVYDSWHVCLP(1)[14.74] ATVRNRLPIPLL(1)[4.25] APPVPLRYLQRL(1)[3.25] APMVLPRSSVSL(1)[27.62] APIIGPKTWLVS(1)[8.88] APEIDRSYLFLF(1)[20.30] ALALPPFPHGYV(1)[12.66] AKRYGIAYEMYV(1)[6.83] 185 Phage display (common panning) 5 PMID:28890361 SH3 domain Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA They validated most of the non-canonical specificity motifs by isolating individual clones from the phage pools to test for positive interactions with the cognate SH3 domains by phage ELISAs. Clonal ElISA signal is the ratio between protein and GST. Qualitatively, everything above 3 is considered a positive binder at uM or lower range. Based on ~70,000 unique binding peptides, we obtained 154 specificity profiles for 115 SH3 domains, which reveal that roughly half of the SH3 domains exhibit non-canonical specificities and collectively recognize a wide variety of peptide motifs, most of which were previously unknown. Crystal structures of SH3 domains with two distinct non-canonical specificities revealed novel peptide-binding modes through an extended surface outside of the canonical proline-binding site. Our results constitute a significant contribution toward a complete understanding of the mechanisms underlying SH3-mediated cellular responses. 3277 CDAGRKQKC(952)[0.48,0.65] CAKRNGKSC CGRKNVKLC CGRKQSERC CQGKRQTEC CGGRKQTAC CAPAGKKQC CYMGKKNAC CGLRGRRQC CKDMGKRNC CDRGRRNPC CGGTARRNC CARRNTRAC CAKKNATTC CSARKQKDC CVSAGKRQC CRYAKKNEC CRGRRNDEC CVGNAKKQC CGKRNARSC CIPGRKNPC CGGRRNRTC CAGKRQPMC CGARRNKSC CVARRNVGC CGRRNC [AG]-B-B-[NQ] 26 Phage display (in vivo) 1 PMID:29123083 Connective tissue growth factor (CTGF) Not determined. Not determined. Not determined. T7 CX7C phage display library ELISA To demonstrate the specific binding of DAG peptide to its receptor, ELISA plate wells were coated with full-length recombinant human CTGF/CCN2 (R&D systems; cat#9190-CC) at a concentration of 5 μ/ml for 2–3 h at room temperature.The reaction was stopped with the addition of equal volume of 1 M sulphuric acid and absorbance measured at 450 nm.(20um,60um) A CX7C naïve phage library (1e10 pfu of, in 100 µL of PBS) was intravenously injected into hAPP-J20 mice and allowed to circulate for 30 minutes due to its short half life, after which mice were anesthetized with 2.5% avertin and perfused with PBS intracardially. The brain was removed and the hippocampus was extracted and homogenized in LB-NP 40 (1%). The phages in the lysate were rescued by amplification in E.coli. 3278 MSSRSRPHINSL(3) TAELYPDLQSSQ(2) EDARRPPTSTEH(2) PVNKQHTSLQNN(2) VNHEYKLHSIKY(1) DDTYPSRPVYLK(1) DLYLSHGAPPQH(1) HVTHNITNESNS(1) ARMTFSQMSPHT(1) TGSIRPKLHASP(1) QLARMSSLHVPM(1) SHEISRITAVSK(1) VDMVTKQLLEYP(1) ELQIGSWRMPPM(1) SERLMTPPKLFR(1) MTHKQMHKHHGL(1) LVSLTPPWINVD(1) SSAQMNLNTFLN(1) LGSHNIRLGEGS(1) YPHPIRQNFFAY(1) KSHTENSFTNVW(1) KLPPMNSDSMVW(1) HMNAHLTFQSAI(1) DAVKTHHLKHHS(1) 24 Phage display (competitive panning) 3 PMID:29119720 1-deoxy-D-xylulose-5-phosphate synthase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) For the first two rounds, DXS functionalized with N-hydroxysuccinimide (NHS)–desthiobiotin and Dynabeads MyOne Streptavidin C1 (Invitrogen 65001) were used to capture DXS from the solution. For the third round, to avoid selection of phages against streptavidin and given that D. radiodurans DXS contains an N-terminal Histag, MagneHis Ni Particles (Promega V8560) were used as capturing system. After the incubation step, 0.1 mL of beads were added to the solution, which was mixed in a thermo shaker at 48Cfor15 min. The tube was placed in a magnetic rack to allow for adhesion of the magnetic beads on one side of the tube. Phages expressing DXS binders were retained on the bead surface, and the buffer containing unbound phages was gently discarded from the tube. To remove weakly bound phages further, the beads were washed with PBST (10V1 mLPBS with 0.05% Tween 20) whilst retaining them using a magnetic rack. The elution of strongly bound phages from the beads was achieved by suspending the beads in the elution buffer (1 mL, 1mM Biotin in PBS for rounds 1 and 2 and 500 mm imidazole in PBS for round 3). 3279 MAIPTRGKMPQY(8) HSSPVQTDWITV(4) KAIRTRGKRPQY(1) YSSTIYTPTAVG(1) GSSLLYSGSGPA(1) ALWPPNLHAWVP(1) SSSPVAWALAMR(1) HSSPPFPWLLVT(1) DSSSGLYLRPLS(1) VSSSIFPIALPD(1) THPSTKVPGTPA(1) ASSVISPRWLLW(1) ALWPPNLHAWVP(1) TSSAAAPYYSPP(1) VSSMKGPTLSTN(1) DSSTWLFLSSYR(1) 16 Phage display (competitive panning) 2 PMID:29119720 1-deoxy-D-xylulose-5-phosphate synthase Not determined. Not determined. Not determined. XS2X9, Ph.D.-12 and wild-type phage library pool Two rounds of selection were performed by using a custom-made phage library. TBS (trisbuffered saline, 50 mm Tris·HCl pH 7.5, 150 mm NaCl) was used as incubation buffer,TBST (TBS with 0.05% Tween 20) was used as washing buffer, MagneHis Ni Particles (Promega V8560) were employed for protein recovery, and 1mm ThDP in TBS was used as elution buffer. ThDP was chosen as competitive eluent to elute peptides specifically interacting with the ThDP-binding site of DXS. A D-GAP-competitive peptidic inhibitor, P12 (MAIPTRGKMPQY), with a Ki value of (113 ± 4) μM, was identified by phage display targeting the thiamine diphosphate (ThDP)-binding pocket. 3280 CSPGAKVRC(322/143637) CGEKRTRGC(279/143637) CDSRSRKRC(226/143637) CPKGAGSKC(196/143637) CRKGTLGRC(149/143637) CVKGTVKTC(148/143637) CRGTIRGRC(143/143637) CLDRRKKPC(136/143637) CPARRSNRC(117/143637) CKDSAEMRC(113/143637) CERSNNVSC(113/143637) CGKRAPFRC(110/143637) CVSSSSGRC(108/143637) 13 Phage display (in vivo) 1 PMID:29116108 Peritoneal macrophage of mouse bearing breast cell line 4T1 Not determined. Not determined. Not determined. T7 CX7C phage display library Naïve phage library was injected intraperitoneally in 4T1 tumor-bearing mice, and allowed to circulate for 2 h. Peritoneal cells were collected, the accompanying phages were rescued and the peptide-encoding segment of phage DNA was sequenced. Tumor-associated macrophages (TAMs) expressing the multi-ligand endocytic receptor mannose receptor (CD206/MRC1) contribute to tumor immunosuppression, angiogenesis, metastasis, and relapse. Here, we describe a peptide that selectively targets MRC1-expressing TAMs (MEMs). Deep sequencing of the peptide-encoding inserts in the selected phage pool revealed enrichment of the peptide CSPGAKVRC (codenamed “UNO”). Intravenously injected FAM-labeled UNO (FAM-UNO) homed to tumor and sentinel lymph node MEMs in different cancer models: 4T1 and MCF-7 breast carcinoma, B16F10 melanoma, WT-GBM glioma and MKN45-P gastric carcinoma. Fluorescence anisotropy assay showed that FAM-UNO interacts with recombinant CD206 when subjected to reducing conditions. Interestingly, the GSPGAK motif is present in all CD206-binding collagens. FAM-UNO was able to transport drug-loaded nanoparticles into MEMs, whereas particles without the peptide were not taken up by MEMs. In ex vivo organ imaging, FAM-UNO showed significantly higher accumulation in sentinel lymph nodes than a control peptide. This study suggests applications for UNO peptide in diagnostic imaging and therapeutic targeting of MEMs in solid tumors. 3281 CTVAGSGPC(77/197903) CIGNSKKKC(66/197903) CVRLREKRC(63/197903) CRKKSRAQC(60/197903) CGKRAPFRC(59/197903) CAANDYSDC(57/197903) CLEDDSAKC(57/197903) CNDKS(55/197903) CVGRKVRGC(55/197903) CVDRRESRC(53/197903) CRQEEESRC(52/197903) CGKTRYSRC(50/197903) CSPGAKVRC(50/197903) 13 Phage display (in vivo) 1 PMID:29116108 Peritoneal macrophage of healthy mouse Not determined. Not determined. Not determined. T7 CX7C phage display library Naïve phage library was injected intraperitoneally in eight-week old Balb/c mice (Charles River, Wilmington, MA, USA), and allowed to circulate for 2 h. Peritoneal cells were collected, the accompanying phages were rescued and the peptide-encoding segment of phage DNA was sequenced. 3282 CRTLRSKAC(775) CRRTRQRSC(500) CIGNK(420) CVLNESGDC(216) CRDKRGSKC(194) CVGRKVRGC(193) CKRPNENVC(183) CNRRTKIGC(182) CSPKMRATC(180) CKRTRRREC(176) CLSSITPEC(169) CVDQDPL(163) CRGTRSNRC(157) CGPCQEGLC(153) CMTLRSRKC(147) CSTKTSLKC(146) CGDEA(146) CTTSTGADC(141) CSTLKRRVC(136) CRGVAKVRC(133) CSVGRLK(131) CHQDF(128) CSFDEANPC(124) CRNRA(123) CVSDRKVAC(123) CKRGRFAKL(120) CAQPNSR(115) CRPTRRVSC(114) CRNGLNKRC(113) CGFRSD(112) CRKTVGPRC(112) CEVMQRKRC(107) CVASVKRKC(106) CDANQ(106) CRRTAIKKC(106) CDSRSRKRC(105) CLSKRTPRC(105) CVDSEATDC(105) CPRTAKVLC(105) CQSRSPRNC(101) CNKNGTAPC(101) CTDRHSTNC(94) CDALAPNSC(93) CIDGRTDLC(92) CMNVESSPC(91) CREKNSQRC(91) CLVRPERKC(90) CRKRMNRTC(90) CVDITSPDC(89) CSYEKEPVC(86) CSPGAKVRC(10) 51 Phage display (common panning) 1 PMID:29116108 Mouse macrophage cell line Raw 267.4 Not determined. Not determined. Not determined. T7 CX7C phage display library 3283 CNDAPLTEC[0.105±0.015] CYSANASSC[0.121±0.012] CGPLPHRHC[0.095±0.045] CNPHFYKHC[0.059±0.006] CTESPNPLC[0.062±0.009] CSMPLMGHC[0.080±0.012] CTMREQHFC[0.021±0.007] CNTHEKPTC[0.060±0.015] CQNTHTKQC[0.195±0.013] CGGSMLSSC[0.133±0.002] CWASNVQSC[0.110±0.007] CTMSHENTC[0.098±0.006] CDSKHVPFC[0.045±0.003] CGSAPVRSC[0.048±0.003] CHMFTGSTC[0.052±0.004] CSKEATPFC[0.058±0.002] 16 Phage display (common panning) 1 PMID:28986145 bB2-crystallin fibrils Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The absorbance of each well was measured at 450 nm using FlexStation® 3 multimode plate reader.Data were reproduced from Figure 6 and shown. 3284 CNAGHLSQC[0.081±0.010] CDFRIRNAC[0.084±0.031] CNTQMHPKC[0.135±0.046] CKQEDILRC[0.131±0.009] CETSNTKSC[0.325±0.042] CGKHQFPHC[0.205±0.051] CGPTAKYIC[0.288±0.052] CSKEATPFC[0.295±0.032] CVNNADNSC[0.125±0.022] CPKGDENTC[0.198±0.020] CVGREPHAC[0.190±0.005] CNVWRAPGC[0.245±0.002] CIKHKAWKC[0.195±0.040] 13 Phage display (common panning) 1 PMID:28986145 bB2-crystallin fibrils Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The absorbance of each well was measured at 450 nm using FlexStation® 3 multimode plate reader.Data were reproduced from Figure 6 and shown. 3285 CKQFKDTTC[0.695±0.001] CVPSKPGLC[0.585±0.121] CFTSTLSRC[0.562±0.112] CMLPTRIAC[0.571±0.041] CDAHYEKSC[0.312±0.052] CNKHNMLNC[0.173±0.031] CYFHYPQRC[0.758±0.110] CSLFSKNYC[0.605±0.035] 8 Phage display (common panning) 1 PMID:28986145 bB2-crystallin fibrils Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The absorbance of each well was measured at 450 nm using FlexStation® 3 multimode plate reader.Data were reproduced from Figure 6 and shown. 3286 YKRGYK(50)[10 ± 2, 620 ± 60] WPQGPT(11)[NT, NT] ETWGYK(6)[NT, NT] TRSGPT(3)[NT, NT] RNSGWS(3)[NT, NT] ARAGTT(2)[NT, NT] 6 Phage display (common panning) 3 PMID:29050803 Platinum Not determined. Not determined. Not determined. T7 LARFH gene-based phage display library (X3GX2) Quartz crystal microbalance (QCM) The interaction of YKRGYK with a platinum surface was monitored and quantified by usinga hand-made flow-quartz crystal microbalance system (f-QCM) with a custom-made acryl flow cell equipped with 9-MHz Pt or Au quartz crystals (Kyocera, Japan). The dissociation constant (KD, nM) was shown. The majority of the selected variants contained the Tyr-Lys-Arg-Gly-Tyr-Lys (YKRGYK) sequence in their randomized segment. We characterized the platinum-binding properties of mutant LARFH by using quartz crystal microbalance analysis. Mutant LARFH seemed to interact with platinum through its loop containing the YKRGYK sequence, as judged by the estimated exclusive area occupied by a single molecule. Furthermore, a 10-residue peptide containing the YKRGYK sequence bound to platinum with reasonably high affinity and basic side chains in the peptide were crucial in mediating this interaction. In conclusion, we have identified an amino acid sequence, YKRGYK, in the loop of a helix-loop-helix motif that shows high platinum-binding affinity. This sequence could be grafted into loops of other polypeptides as an approach to immobilize proteins on platinum electrodes for use as biosensors among other applications. 3287 CSLFKNFRC CRADFYTTC CRENTNHTC CWNEDHTWC CLNSLFGSC CTYSPTEVC CQLFPLFRC CTLQDQATC CMQHSMRVC 9 Phage display (common panning) 1 PMID:29103911 Mouse heart Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) For in vitro evaluation of uptake by skeletal and cardiac cells, we synthesized the best 12 candidate peptides equipped with a fluorescein isothiocyanate (FITC) label we used the Ph.D.-C7C phage display peptide library. A schematic outline of the screening procedure is given in Figure 1. We performed a first-round in vitro screening using human control myotubes in which internalized and surface-bound phages were isolated and amplified. Subsequently, the naive library was also used for a first-round in vivo enriched internalized phage fraction, and the enriched surface-bound phage fraction was used for a second in vivo screening round in mdx mice, a mouse model for DMD.All enriched libraries from the biopanning selections, and the naive library with and without one round of bacterial amplification, were sequenced using a published NGS approach. 3288 VHWDFRQWWQPS(5)[1.04±0.08] GDGNSVLKPGNW(1)[0.49±0.07] HLSLPLWKWEKS(1)[0.61±0.09] VFAYHLRIPSGD(1)[0.43±0.13] 4 Phage display (common panning) 3 PMID:29163684 The second extracellular loop (107-130) of C-C chemokine receptor type 9 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Affinity was detected using ELISA. The OD450 values were reproduced from Figure 1 in the literature. P1 was (VHWDFRQWWQPS) able to bind specifically with MOLT4 cells which exhibit marked expression of CCR9. In addition, P1 promoted the apoptosis of MOLT4 cells induced by doxorubicin, and inhibited the migration of MOLT4 cells in the presence of chemokine (C-C motif) ligand 25. It was suggested that decreased activity in the phosphorylation of protein kinase B in MOLT4 cells may be responsible for the inhibition. In conclusion, the peptide P1 derived from a screened phage is able to specifically bind to CCR9 and inhibit the activity of CCR9. It has potential use as an antagonist in the treatment of CCR9-overexpressed carcinoma. 3289 CQVRSNTTC CSLFKNFRC CWNEDHTWC CLNSLFGSC CLLGHTNNC CFSHTYRVC CQLFPLFRC CMQHSMRVC 8 Phage display (common panning) 1 PMID:29103911 Mouse muscle Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) we used the Ph.D.-C7C phage display peptide library. A schematic outline of the screening procedure is given in Figure 1. We performed a first-round in vitro screening using human control myotubes in which internalized and surface-bound phages were isolated and amplified. Subsequently, the naive library was also used for a first-round in vivo enriched internalized phage fraction, and the enriched surface-bound phage fraction was used for a second in vivo screening round in mdx mice, a mouse model for DMD.All enriched libraries from the biopanning selections, and the naive library with and without one round of bacterial amplification, were sequenced using a published NGS approach. 3290 SRRPASFRTARE(15)[68] GLHTSATNLYLH(4)[NT] 2 Phage display (common panning) 4 PMID:29152066 Extracellular domain of Fibroblast growth factor receptor 2 [1-377] Not determined. 1EV2,1II4,1IIL, Not determined. Ph.D.-12 phage display library (X12) Flow Cytometry The Cy5.5-labeled peptide was serially diluted in PBS at concentrations ranging from 0 to 200 nM in 25 nM increments. QhTERT/FGFR2c cells (~1.0e5) were incubated with peptide at 4°C for 1 hour, washed with cold PBS, and the mean fluorescence intensities were measured using flow cytometry. The equilibrium dissociation constant kd=1/ka was calculated by performing a least squares fit of the data to the non-linear equation I=(I0+Imaxka[X])/(I0+ka[X]). I0 and Imax are the initial and maximum fluorescence intensities, corresponding to no peptide and at saturation, respectively, and [X] represents the concentration of the bound peptide. Prism 5.0 software (GraphPad Inc) was used to calculate kd (nM). Four rounds of biopanning were performed using a decreasing quantity (100, 80, 60, and 40 μg) of FGFR2-ECD in successive rounds to increase binding specificity. We used phage display to identify the peptide SRRPASFRTARE that binds specifically to the extracellular domain of FGFR2. We labeled this peptide with a nearinfrared fluorophore Cy5.5, and validated the specific binding to FGFR2 overexpressed in cells in vitro. We found high affinity kd = 68 nM and rapid binding k = 0.16 min-1 (6.2 min). In human esophageal specimens, we found significantly greater peptide binding to high-grade dysplasia (HGD) versus either Barrett’s esophagus (BE) or normal squamous epithelium, and good correlation with anti-FGFR2 antibody. We also observed significantly greater peptide binding to excised specimens of esophageal squamous cell carcinoma and gastric cancer compared to normal mucosa. These results demonstrate potential for this FGFR2 peptide to be used as a clinical imaging agent to guide tissue biopsy and improve methods for early detection of esophageal adenocarcinoma (EAC) and potentially other epithelial-derived cancers. 3291 TWPKHFDKHTFYSILKLGKH[45] 1 Phage display (common panning) 6 PMID:29090274 Domain 3332-3779 of prolow-density lipoprotein receptor-related protein 1 Alpha-2-macroglobulin receptor-associated protein, Alpha-2-MRAP 2FYL, Not determined. T7 X12, X16 and X20 phage display library pool ELISA Binding activity and selectivity of biotinylated TWPKHFDKHTFYSILKLGKH (L57) to LRP1(3332-3379)-Fc were evaluated by plate ELISA. Binding activity was detected with HRP-conjugated streptavidin and was measured by determining the absorbance at 450 nm. LRP1(3332-3379)-Fc and Act RIIB-Fc are counter proteins. The absorbance at 450 nm of biotinylated L57 was expressed as the mean ± SD (n = 2). The EC50 (nM) value of L57 for binding to LRP1(3332-3379)-Fc was shown. For the screening of T7 phage libraries, 20 μg LRP1-Fc was immobilized on Dynabeads Protein A (200 μL; Invitrogen, Carlsbad, CA, USA) in PBS (045–29795; Wako, Osaka, Japan) containing 0.5% BSA. After washing of the beads with PBS containing 0.1% Tween-20 (PBST), they were incubated with 1.0e12 pfu of phage libraries (mixture of X12, X16, and X20) for 1h at room temperature, and then washed with PBST (three times for the first round, five times for the second round, and 10 times for the third to sixth rounds). Bound phages were eluted with 200 μL of 0.5% SDS and then transfected into 80 mL of E. coli BLT5615 cells (Merck Millipore) in the log phase of growth for amplification. After bacteriolysis, the phages were recovered from the culture supernatant by centrifugation and PEG precipitation, according to the T7 Select manual. The recovered phages were dissolved in 1mL of PBS and 0.5 mL of the phages was used for the next round of biopanning. Using phage display technology, we obtained a novel peptide, L57 (TWPKHFDKHTFYSILKLGKH-OH), with an EC50 value of 45 nM for binding to cluster 4 (Ser3332–Asp3779) of LRP1. L57 was stable in mouse plasma for up to 20 min. In situ brain perfusion assay in mice revealed the significantly high BBB permeability of L57 3292 SVDLSSFIPPL(1)[2.97 ± 0.09] AAASLRDFISRW(1)[3.53 ± 0.14] DYGIWGRWIPLI(1)[3.19 ± 0.02] GYNLRAWLPPPY(1)[3.24 ± 0.07] VPVHLGSWLPPP(1)[3.34 ± 0.07] WSPLSLTRFIPP(1)[3.41 ± 0.01] FDPWLTKYIPYP(1)[3.54 ± 0.07] ELPRLNQYIPPP(1)[3.68 ± 0.07] GAMHLPWHMGTL(10)[1.64 ± 0.11] GAMHLPWHMGTP(1)[1.68 ± 0.07] TSMKLDRWIPPL(1)[3.37 ± 0.01] WNPMVNFIEPPH(1)[2.75 ± 0.08] GPFVYNQWIPIP(1)[3.36 ± 0.02] FLTDYIPTPSRK(1)[3.10 ± 0.07] VNMTQFVPPPAR(1)[3.50 ± 0.15] YSTNYTAFIPGP(1)[3.50 ± 0.13] EVNLSRFIQLPS(1)[2.70 ± .018] WEGRFLHFLNYP(1)[2.65 ± 0.24] 18 Phage display (common panning) 3 PMID:29310289 Pesticidal crystal protein Cry1Ab Anti-Cry1Ab monoclonal antibody Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA 100 μL of Cry1Ab protein was coated in microtiter plate wells and incubated overnight at 4 °C. After blocking with 3% skimmed milk at 37 °C for 2 h and washing 3 times with PBST, 100 μL of each serial concentration of selected phage stock was added to the Cry1Ab coated wells and incubated at 37 °C for 1 h. Following washing 3 times with PBST, 100 μL of HRP-conjugated anti-M13 phage antibody (1:5000, diluted in PBS) was added to the wells and incubated at 37 °C for 1 h. Then, 100 μL 3,3′,5,5′-tetramethylbenzidine (TMB) substrate solution was added to each well and incubated for 10 min at room temperature with gentle agitation. The optical density at 450 nm was detected on microplate reader (Thermo Scientific). Variety of phage display peptides capable of binding to Cry1Ab were obtained by phage display peptide library panning, in which Ab-39(TSMKLDRWIPPL) not only had good binding activity to Cry1Ab but also highly specific for Cry1Ab. After modification of electrode with gold nanoparticles, selected phage displayed peptide was applied to electrochemical immunosensor for Cry1Ab. 3293 ATRSANGM[6.18 ± 0.48] EQVPYPGE[0.29 ± 0.07] ASRSPTDK[0.27] ASRGTIAG[3.09] 4 Phage display (common panning) 3 PMID:29414074 Prostate-specific antigen Anti-human total PSA monoclonal antibody Not determined. Not determined. f8-8mer phage display library (X8) ELISA The phage was loaded into wells of a 96-well microtiter plate, and 50 μL of t-PSA solution was added to the wells, and the plate was incubated at 37 °C. Then the wells were washed six times with PBST, anti-PSA mAb was added. The plate was washed again, and then 50 μL of IgG-HRP was added and the plate was incubated for 1 h at room temperature. For the detection of t-PSA, 100 μL of 0.04% OPD was added. The absorbance at 492 nm was recorded on a microplate reader. ~10e11 phage virions from f8/8 landscape phage library in 50 μL library diluents were added to the wells as the input for the first round of biopanning. After incubation overnight at 4 °C, the wells were washed six times with TBST to remove unbound phages. After three rounds of selection, individual phage clones were picked randomly and amplified. Peptide ATRSANGM showed highest affinity and specificity against t-PSA. 3294 GTQYYAYSTTQKS GMIVDHLPIQVNT GEYDYACGVVGYE GTQAIRVHTISSQ GSYPKASLALLAP GGLNQVLRIPSFI GSPKHNLDMVKMM 7 PMID:29493687 MgF2 nanoparticles (NPs) Not determined. Not determined. Not determined. GX12 phage display library Biopanning on MgF2 nanoparticles (NPs) in water provided recently seven different 12mer peptides and one of which was selected to be investigated as peptide–poly-(ethylene glycol) (pep–PEG) conjugate with a PEG-block of Mn = 3000 (pep-I-PEG). 3295 CFNLALHTC CISHGSPQC CLSTASNSC 3 Phage display (subtractive panning) 4 PMID:29516383 EAE-brain capillary endothelial cell Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Brain endothelial cells derived from four deeply anesthetized rats with acute EAE (score 3) and four healthy rats were isolated. Briefly, brain pieces were gently homogenized, mixed with 30% dextran. The vascular fraction was filtered and digested in DMEM. Endothelial cells were pelleted and incubated with the phages for 2 h at 20 °C. The counter-selection step was performed by incubating 5e5 healthy brain capillary endothelial cells (BCECs) with 1e11 phages in 1 mL RPMI. After centrifugation of cells, the unbound phages were transferred on 4e5 EAE-BCEC for 2 h to obtain phages of the EAE-BCEC selection. The phages that remained bound to cells were recovered, amplified in E. coli ER 2738 bacteria. Four selection rounds were performed with isolated healthy or EAEBCEC. These peptides were respectively predicted to mimic the discointeracting protein 2 homolog A (DIP2A), the carboxypeptidase N, polypeptide 2 (CPN2), a plasma zinc metallocarboxypeptidase important in regulating vasoactive peptide hormones, growth factors and cytokines by specifically cleaving their C-terminal basic residues (Ph37), the intracellular protein huntingtin (Ph38), and the laminin subunit-1, a component of the extracellular matrix, in a region of epidermal growth factor (EGF)-like domain (Ph48). 3296 CSYKARVTC 1 Phage display (subtractive panning) 5 PMID:29516383 EAE-CNS brain endothelial cell Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) CNS cells were prepared from rat CNS. A rat with acute EAE (score 3) was deeply anesthetized and perfused with 50 mL ice-cold PBS 1×. The whole CNS tissues from both rats were recovered separately (about 2.3 g), cut into small pieces, and incubated for 50 min at 37 °C in 10 mL of PBS 1×. The recovered cells were concentrated and frozen. The Bex vivo total EAE selection^ was performed in two steps: (i) a counter-selection with CNS cells from a healthy rat realized to absorb healthy tissuespecific phages and (ii) the selection against cells from the EAE CNS. Thus, 1011 phages were incubated with 4.0 × 106 CNS cells from a healthy rat before incubation of the unbound phages with CNS cells from a EAE rat (4.0 × 106). After incubation, the cells were pelleted by centrifugation (1200 rpm/5 min/RT) and washed with 1 mL complemented RMPI. The phages that remained associated with cells were amplified. At each selection round, one additional washing step of cells associated with phages was added to increase the selection stringency, so that five washes were performed during the fifth round. This peptide mimics the discointeracting protein 2 homolog A (DIP2A), receptor for Follistatin 1, a glycoprotein involved in inflammatory reactions (Ph36). 3297 ACNTWNPWCPWDAPLCA[2.3 ± 2.7][0.40 ± 0.24] ACNNFPFRCVYYPDICA[0.44 ± 0.07][NT] 2 Phage display (common panning) PMID:29517911 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. CX6CX6C phage display library Inhibition assay NaCl, 1.5 μM bovine serum albumin, pH 7.5. PKal (typical nominal concentration 1−2 nM) was preincubated in 96-well plates with different concentrations of the tested peptide (or of aprotinin, used as a control molecule) for 15 min. The maximum final concentration of a given peptide was chosen based on its potency and was followed by a 1.25- to 3-fold serial dilution, plus a control with no peptide. The reactions were initiated by the addition of H-Pro-Phe-Arg-AMC at a final concentration of 10 to 25 μM, and initial rates of substrate hydrolysis were determined by linear fit of the raw fluorescence versus time traces. In the case of H-Pro-Phe-Arg-AMC however, the Km for PKal is large, and all experiments were performed under conditions where [S]≪ Km. The affinity of peptide binding to human and rat PKal are showed respectively. Bicyclic peptides with inhibitory activity against PKal were identified using Bicycle Therapeutics proprietary phage display platform. Briefly, linear peptide libraries containing randomized AA between 3 cysteine residues were displayed on the surface of filamentous phage and cyclized on the phage with a thiol-reactive molecular scaffold (TBMB). The libraries formats used were 5 × 5 and 6 × 6, such that either 5 or 6 randomized AA were between the first and second and second and third cysteine, respectively. Following cyclization of the phage libraries, repeated selections were performed against human and rat PKal. Selections and subsequent affinity maturation selections were performed against PKal using methodology described previously. Rat and human PKal orthologous enzymes were alternated as baits during each selection round in order to identify bicyclic peptides with good cross-reactivity. peptides were chemically synthesized for characterization. 3298 ACSWPARCLHQDLCA[0.07 ± 0.02][0.90 ± 0.33] 1 Phage display (common panning) PMID:29517911 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. CX5CX5C phage display library Inhibition assay NaCl, 1.5 μM bovine serum albumin, pH 7.5. PKal (typical nominal concentration 1−2 nM) was preincubated in 96-well plates with different concentrations of the tested peptide (or of aprotinin, used as a control molecule) for 15 min. The maximum final concentration of a given peptide was chosen based on its potency and was followed by a 1.25- to 3-fold serial dilution, plus a control with no peptide. The reactions were initiated by the addition of H-Pro-Phe-Arg-AMC at a final concentration of 10 to 25 μM, and initial rates of substrate hydrolysis were determined by linear fit of the raw fluorescence versus time traces. In the case of H-Pro-Phe-Arg-AMC however, the Km for PKal is large, and all experiments were performed under conditions where [S]≪ Km.The affinity of peptide binding to human and rat PKal are showed respectively. Bicyclic peptides with inhibitory activity against PKal were identified using Bicycle Therapeutics proprietary phage display platform. Briefly, linear peptide libraries containing randomized AA between 3 cysteine residues were displayed on the surface of filamentous phage and cyclized on the phage with a thiol-reactive molecular scaffold (TBMB). The libraries formats used were 5 × 5 and 6 × 6, such that either 5 or 6 randomized AA were between the first and second and second and third cysteine, respectively. Following cyclization of the phage libraries, repeated selections were performed against human and rat PKal. Selections and subsequent affinity maturation selections were performed against PKal using methodology described previously. Rat and human PKal orthologous enzymes were alternated as baits during each selection round in order to identify bicyclic peptides with good cross-reactivity. peptides were chemically synthesized for characterization. 3299 PGATAGPSS[1.47 ± 0.04] STRRIGWLTD[1.50 ± 0.03] TDQTHRLA[1.46 ± 0.05] PGATAGPSS[1.55 ± 0.04] PGATAGPSS[1.38 ± 0.04] TDQTHRLA[1.42 ± 0.07] STRRIGWLTD[1.46 ± 0.03] SWVHEWVTSN[1.56 ± 0.01] TDQASRLA[1.46 ± 0.03] RSGCTSGLHR[1.51 ± 0.04] TDQASRLA[1.45 ± 0.03] RSGCTSGLHR[1.43 ± 0.03] 12 Phage display (common panning) 3 PMID:29522802 Anti-S.aureus GapC monoclonal antibody(mAb2A9) Glyceraldehyde-3-phosphate dehydrogenase Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Detection of the binding activities of 12 strongly positive phage clones to mAb2A9 that were selected by a sandwich ELISA and the OD450 value of each well was read using a microplate reader (Bio-Rad, Hercules, CA, USA). Wild-type M13 phage was used as the negative control and an E. coli ER2738 culture supernatant was used as the blank control. Bovine serum albumen-coated wells were used to exclude cross-activity. The consensus motif PVATGSLT of the 12 clones is located at amino acids 236–243 of the GapC protein. These results suggest that the motif 236PVATGSLT243 is an epitope of the S. aureus GapC protein and it might be a potential component of an epitope-based vaccine against S. aureus. 3300 VQTVQIGSD[0.34±0.02] VSSVSTGES[0.15±0.02] VSTLESNQD[0.14±0.01] 3 Phage display (common panning) 3 PMID:29362753 Vibrio parahaemolyticus Not determined. Not determined. Not determined. f8-9mer phage display library (X9) Dot blot The specificity and affinity of the phages binding to V. parahaemolyticus were determined by the phage capture assay. A V. parahaemolyticus specific fusion phage ligand was discovered from the f8/9 landscape phage library through a biopanning procedure. The phages from the f8/9 landscape library were added to a well of a 96-well plate with immobilized V. parahaemolyticus. After washing to remove the unbound phages, the V. parahaemolyticus binding phages were eluted, propagated, and purified for use as the input in the next two rounds of biopanning. After 3 rounds of biopanning, we randomly picked up 5 phage clones for PCR amplification and sequencing. The frequency of the phage VQTVQIGSD was higher than that of others. 3301 YSLHNAGPWSLQ[0.29] 1 Phage display (common panning) 3 PMID:29433534 Immunoglobulin G(IgG) of patients with Rheumatoid arthritis-Secondary Sjögren’s syndrome(RA-sSS) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Antibody reactivity against peptides was measured using a solid-phase ELISA and the color intensity was measured at 450 nm. Phage clones with the sequence YSLHNAGPWSLQ specifically bound to the IgGs of patients with Rheumatoid arthritis-Secondary Sjögren’s syndrome(RA-sSS), which was confirmed by three rounds of screening with the same pooled IgGs. 3302 ARGRRG GRSGRG ARGRGR RSGRAG ARGGGR RSGGRG RGRSGG GRGARG AVGRGR ARGGRG VGRSRG RVRGRG RGRGGR VGRARG GRAAGR SVGRAR ARGRLG GRSGGR RGARAG PRGRRG 6722 Phage display (common panning) 1 PMID:29434246 Thrombin from prothrombin[328-622] Not determined. Not determined. Not determined. X6 phage display library Out of 5.3 × 106 unique peptide sequences identified following thrombin selection, 6722 peptides were significantly enriched, and identified. Analysis of selected phage sequences confirms a general preference for Arg and exclusion of acidic amino acids in the cleaved peptides. Arg was the dominant amino acid within the most significantly cleaved peptides, consistent with the known requirement at P1 of thrombin substrates. 3303 AVLGGG LGSVAE REWGAL SVRWVG HGLSAV ALTVAG AWMRLG GLAAVR GWGAVR SVAAWR MGRWGV TSSVSL ASVMVV LSTAVG VVAGGV TLVRVA VVGGWG AVRLSV GWGVAG RLVWGA 95 Phage display (common panning) 1 PMID:29434246 A disintegrin and metalloproteinase with thrombospondin motifs 13(ADAM-TS 13) Not determined. Not determined. Not determined. X6 phage display library Compared with thrombin, ADAMTS13 appears to exhibit narrow substrate specificity, since VWF is its only known substrate11. Consistent with this observation, only 96 cleaved peptides were identified from the random peptide library following overnight selection by ADAMTS13. Although cleaved peptides preferentially contained bulky hydrophobic amino acids, no obvious motif was observed, consistent with previous studies that demonstrate poor recognition of short peptidyl substrates by ADAMTS13. 3304 LELYLS IQLFLA LRYSSM LVYMHG LVPFLS LLGYTA RLRYFL IMMFLG LVTYVS PRYYLS LAYSRN LIMLLL LRHYFM FTLFMV LTYMRL VTYMVT LHYLNY LTFLLF LRYRDS LRFVTR 1670 Phage display (common panning) 1 PMID:29434246 A disintegrin and metalloproteinase with thrombospondin motifs 13(ADAM-TS 13) Not determined. Not determined. Not determined. VWF73(P3-P3′) phage display library(X6) Following treatment of the VWF73(P3-P3′) library with ADAMTS13, 1670 cleaved peptides were detected. Bulky aliphatic amino acids were preferred in the enriched peptide pool, whereas acidic amino acids, as well as proline and cysteine, appeared to antagonize ADAMST13 substrate recognition. The native amino acid sequence for VWF within this interval (Leu-Val-Tyr-Met-Val-Thr) was among the top peptides identified (pFDR = 8 × 10−5), and none of the most significantly cleaved peptides exhibited faster substrate performance than wild type VWF73. As a result, peptides with lower P-values than wild type VWF73 are not necessarily cleaved more efficiently. 3305 GDALFSVPLEVY SIDDQRDVAEFA TLHQPPSSANWI PIDDQRDLAEFA FTPGGNTYAGQP SIDDQRDVGEWG SMCCMTSVSDQP AVSPPSTIWVPN VTADSQSPSNQV 9 Phage display (in vivo) 3 PMID:29435721 Cervical cancer Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Specific phages targeting cervical cancer in vivo were selected with the cervical cancer xenograft model by screening the Ph.D.™-12phage display library for three rounds. 3306 KQNLAEG STLGPLF IQQRGEA 3 Phage display (in vivo) 3 PMID:29435721 Cervical cancer Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Specific phages targeting cervical cancer in vivo were selected with the cervical cancer xenograft model by screening the Ph.D.™-C7C phage display library for three rounds. 3307 VHWDFRQWWQPS(19) LNFPMPSRPHSS(5) QSLPWCYPHCVT(3) WSPISGKFFQRF(2) NQLLTQRTPFQD(1) SSNVISPFEQLL(1) WVPWSTPFWLQQ(1) NSPSGLIGWTSR(1) WVPWSYEYFVST(1) GPYVLGEHLRSN(1) SPLWSGWALEIL(1) SFTWLHGSLTER(1) SNTQNVYWERWI(1) RFPGPIEPDLRF(1) 14 Phage display (common panning) 3 PMID:29447281 Insulin-degrading enzyme Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3308 SWMSIHL(3) SWWNIHL(2) NAGHLSQ(2) NWMNIHM(1) SKNFPRN(1) DWMRIWN(1) IHSPTAL(1) LPWPLSL(1) SWWMIHH(1) LWWQLWL(1) GFTTTFV(1) SWWSIHV(1) LLLLNNT(1) NWWTIHN(1) NSIKKWS(1) ISSSINH(1) 16 Phage display (common panning) 3 PMID:29447281 Insulin-degrading enzyme Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 3309 GVIHRNDQWTAPGGG DRWVARDPASIFGGG SVMNTSTKDAIEGGG 3 Phage display (common panning) 3 PMID:29463859 MoS2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3310 LGDLHNRDNNSA(1) HSRTDYVQASYP(1) GLHTSATNLYLLH(5) GDGNSVLKPGNW(1) TASDVPRSRPHS(1) SGVYKVAYDWQH(5) 6 Phage display (in vivo) 3 PMID:29473787 Mouse heart Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A total of three pannings were performed. Briefly, at day 21 after induction of EAM, animals received an intravenous injection through the tail vein with either a linear 12 amino acid (AA) peptide phage library or an empty phage at a concentration of 1e9 plaque-forming units (pfu)/μl of PBS.For each panning, animals were divided into the following treatment groups: MyH PhD, MyH Cont,PBS PhD and PBS Cont. The hearts for each of the four treatment groups were weighed, pooled together and homogenized. The resulting homogenate solution for each treatment group was then used to amplify, purify and titer the phages that bound to the hearts in each treatment condition. The phages isolated in panning 1 were injected at 1 × 109 pfu/μl of PBS (110 μl total volume) to animals in the same treatment group in panning 2, and the phages isolated in panning 2 were injected at same concentration and volume to animals in the same treatment group in panning 3. We identified one peptide, MyH-PhD-05, that targets severe myocarditis in vivo. 3311 VHWDFRQWWQPS(15)[1.00±0.05] AWYSNLLPLARF(1)[1.00±0.05] LPAQVGQGPLGT(1)[NT] WFPTSRWTSGWI(1)[NT] VDARYWTRGTYM(1)[NT] AKHPDHPLTVGG(1)[NT] 6 Phage display (common panning) 4 PMID:29046922 Transferrin receptor protein 1 Iron-loaded human transferrin Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The geo mean fluorescence values of gradient concentration samples were fitted to the equation (Y = BmaxX/(Kd + X)), and the Kd was used to evaluate the affinity of these peptides.Compared with these peptides, Y1 and Y2 have the comparable affinity at the micromole range, except peptide THRPPMWSPVWP, the Kd of which is low, indicating that the binding of peptides Y1 and Y2 to CD71 is 20 times higher than their binding to other proteins and 8.7 times higher than their binding to protein-free beads. After four rounds of selection, the bound phages were collected and used for a counter selection. 3312 HSVGNFWSDYSKYLDSRRAQDFVQWLMLT HSQGTFTSDYSKYVEDRRAHDFVQWLMNT HSQGTFTSDYRKYLDERAAWDFVQWLMNT HSQGTFTSDYSKYLDIGRAQDFVQWLLNT HSQGTFTSDYSKYLDSLMAQDFVQWLMST HSQGTFTSDYSKYLDWRRAQDFVQWLLNT HSQGTFTSDYIKLLDSRRAQDFVQWLMNT HSQGTFTSDYSKYLDARRAQDFVQWLIRT HSQGTFTSDYSKYLDVRRAQDFVQWLMNT HSQGTFTSDYSKYLDELRAYDFVQWLMNT HSQGTFTSDYSKYLDSRRAHDFVQWLLNT HSQGTFTSDYSKYLDSRRAQDFVQWLMNP HSQGTFTSDYSKYLDSRRAQDFVQWLINY INHEQWAFTSDYSKYLDSRRAQDFVQWLMNT ASMFTFFSDYSKYLDSRRAQDFVQWLMLT HSQGTFLSDYSKLLDSRRAQDFVQWLMQT HSQGTFLHDYYYYLDSRRAQDFVQWLMDT HSQGTFTSDYSKYLDSIRAQDFVQWLMDT HSQGTFTSDYSKYLDSRRAQDFVDWLMNE HSQGTFTSDYSKYLDSRRAQDFVQWLINT KALGQFTFTSDYSKYLDSRRAQDFVQWLMNT HSQGTFFSDYSHWLDSRRAQDFVQWLMNT HSQGTFTSDYSKYLDWRRAQDFVQWLQNT HSQGTFTSDYSKYLDSKRAHDFVQWLLNT HSQGTFTSDYSKYLDSRRAQDFWIDLMNT HSQGTFTSDYSKYLDSRRAQDFVMTSMNT HSQGTFTSDYSKYLDSRRAQDFVEWLMNN HSQGTFTSDYSKYLDSRRAQDFVDWLINS HSHGTFTSDYSKYLDSRRAQDFVQWLMTT HSQGIFFSDYSKYLDSRRAQDFVQWLMNT HSQGTFTSDYSWYLDSRRAQDFVQWLMNT HSQGTFTSDYSKYLDMQRAHDFVQWLMNT HSQGTFTSDYSKYLDSRMAYDFVQWLMNT HSQGTFFSDYSKYLDSRRAQDFVQWLLET HSQGTFTSDYSKYLDSRRAQDFVQWLLDS 35 Phage display (common panning) 4 PMID:29330364 glucagon receptor(GCGR-) and glucagon-like peptide-1 receptor(GLP1R-)overexpressing cells Not determined. Not determined. Not determined. X7FTSDYSKYLDSRRAQDFVQWLX2T, X7SDYSKYLDSRRAQDFVQWLX2T, HSQGTFX7LDSRRAQDFVQWLX2T, HSQGTFTSDYSKYX7DFVQWLX2T and HSQGTFTSDYSKYLDSRRAQX9 phage display library pool 1e11 phages of the Library Mix were used for the first round of selection on GCGR+ and GLP1R+ overexpressing HEK293 cells, after depletion on naïve cells. Four rounds of selection were carried out, with two different selection schemes: in Scheme 1, the first round was made on GCGR+ cells, followed by a second round on GLP1R+ cells, a third round on GCGR+ cells, and a fourth round on GLP1R+ cells; in Scheme 2, the order of the cells was inverted. Briefly, phage particles were blocked, then incubated with 1e7 naïve cells previously resupended in MPBS for 1 additional hour in a rotary mixer. Cells were incubated with either GLP1R+ or GCGR+ cells, resupended into 0.5 mL of MPBS for 1hr in a rotary mixer. After incubation. The day after the culture was centrifuged at 4200 rpm for 45 min at 4 °C, the supernatant was recovered and used to perform the next round of selection. The Library Mix was selected alternatively on GCGR+ and GLP1R+ HEK293 overexpressing cells in each round. For selection on GCGR+ and GLP1R+ cells, the five libraries were pulled (‘Library Mix’) and selected together. 3313 DSSAVCWAFPHHPLCHMKAT[0.15][4.98] ADADMCWFFPTSPWCH[0.92][0.67 ± 0.62] DLSAVCWAFPWDPECH[0.03][0.016 ± 0.013] DSSAVCWAFPYLPECH[0.17][0.048 ± 0.018] DISAVCWAFPFDPECH[0.25][0.039 ± 0.018] 5 Phage display (common panning) 3 PMID:29329326 Avi-tagged interleukin-17A Not determined. Not determined. Not determined. IX104 phage display library(X5CX8CX5) ELISA,Surface plasmon resonance (SPR) BIAcore T200 was used to determine binding affinity and kinetics of the peptides to IL-17A. Avitagged IL-17A was captured to streptavidin sensor chip.For each SPR run, the sensor was first conditioned by 8 cycles of blank prior to sample injections.The resulting sensorgrams of the concentration series of a same peptide were fitted globally to extract binding kinetics and affinity with BIAcore T200 Evaluation Software 2.0 using a 1:1 binding model. Pure peptides from the two libraries were assayed in AlphaLISA format to investigate the potential for disruption of the IL-17A/IL-17RA cytokine-receptor interaction in a biochemical format. In general, the IC50 values determined by AlphaLISA agreed quite closely with the KD values determined through SPR analysis.As was observed in SPR for this family, the differences in AlphaLISA activity between the affinity matured peptides were not significant, with several peptides having IC50's in the 0.2 to 2.0 μMrange. IX104 phage display library(X5CX8CX5) consisting of 18 random amino acids were selected against 100 nM of Avi-tagged IL-17A. After three rounds of selection were completed, the eluted phage pool was deconvoluted by filter-lift. 3314 AYECPRLEYDMFGALHCLPS[12.30][61.7] CPRLEYDMFGALHCL[4.06][5.3 ± 2.5] CLDLQYDPWGALHCI[2.56][2.0 ± 1.3] CFDLQYDPWGALHCI[2.41][1.5 ± 0.4] CLDLQYDMFGALHCV[3.87][1.9 ± 0.7] CLDLVYDPWGALHCI[4.04][3.1 ± 1.2] CWVLEYDMFGALHCR[1.30][1.9 ± 1.5] CWALEYDMFGYLHCR[0.29][0.72] CWVLEYDMFGFLHCR[0.072][0.20] CWVLEYDMFGYLHCR[0.073][0.19] 10 Phage display (common panning) 3 PMID:29329326 Avi-tagged interleukin-17A Not determined. Not determined. Not determined. IX104 phage display library(X3CX12CX3) ELISA,Surface plasmon resonance (SPR) BIAcore T200 was used to determine binding affinity and kinetics of the peptides to IL-17A. Avitagged IL-17A was captured to streptavidin sensor chip.For each SPR run, the sensor was first conditioned by 8 cycles of blank prior to sample injections.The resulting sensorgrams of the concentration series of a same peptide were fitted globally to extract binding kinetics and affinity with BIAcore T200 Evaluation Software 2.0 using a 1:1 binding model. Pure peptides from the two libraries were assayed in AlphaLISA format to investigate the potential for disruption of the IL-17A/IL-17RA cytokine-receptor interaction in a biochemical format. In general, the IC50 values determined by AlphaLISA agreed quite closely with the KD values determined through SPR analysis.As was observed in SPR for this family, the differences in AlphaLISA activity between the affinity matured peptides were not significant, with several peptides having IC50's in the 0.2 to 2.0 μMrange. IX104 phage display library(X3CX12CX3) consisting of 18 random amino acids were selected against 100 nM of Avi-tagged IL-17A. After three rounds of selection were completed, the eluted phage pool was deconvoluted by filter-lift. 3315 CPDEKHQ[1.63±0.11] CPDEKHQ[1.65±0.05] GLPPREIVD[1.62±0.12] KACPDEKHS[1.63±0.07] TRNTRT[1.68±0.05] QESQEPDIHY[1.75±0.08] GLPPREIVD[1.83±0.04] IGLEMVFQLI[1.86±0.07] INVTKLLIQI[1.72±0.03] QESQEPDIHY[1.65±0.07] TTRNTRTR[1.72±0.09] QRNKAAHSVN[1.75±0.04] GYTEDEIV 12 Phage display (common panning) 3 PMID:29304128 Anti-GapC monoclonal antibody (mAb) 1F4 Glyceraldehyde-3-phosphate dehydrogenase Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Twelve clones showed strong, positive reactions with mAb 1F4. The OD450 of each peptide binding to mAb was read using a microplate reader at 450 nm. 3316 YSLRSDY(5)[0.59±0.02] YSLRQDW(3)[0.37±0.01] YSLRTDW(3)[0.48±0.01] YSLRQER(3)[0.50±0.01] YSLRADR(2)[0.39±0.01] ALSSLRN(2)[0.55±0.02] HSIRYDF(2)[0.62±0.01] HSIRVDW(2)[0.35±0.01] KVWIVPS(2)[0.50±0.01] KVWLLHS(2)[0.49±0.02] DWIFPAF(1)[0.43±0.02] 11 Phage display (common panning) 4 PMID:29301517 Glycoprotein G Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA ELISA plates were coated with the G1 protein and the purified pET32a(+) vector control protein coated with the same concentration. The absorbance of the reaction was determined at 450 nm with an ELISA microplate reader. 27 specific peptide ligands displaying 11 different amino acid sequences were obtained, and the HSIRYDF and YSLRSDY clone had a higher affinity to G1 protein than the other clones. 3317 IMPHKHRRKLRL(1)[0.50 ± 0.02] MLTISQRQLSIS(1)[0.48] RNRTKKQITTQR(1)[0.56 ± 0.01] RMKMLMMLMRRK(11)[0.52] 4 Phage display (common panning) 3 PMID:29517966 Anti-RhD monoclonal antibody Blood group Rh(D) polypeptide Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Plates were coated with monoclonal anti-D at 4 °C overnight. Phage clones were added to the antibody-coated wells and incubated for 1 h at room temperature. The primary phage library was used as the negative control The optical density (OD) at 450 nm was subsequently recorded using a microplate reader. In the second and third rounds of biopanning, 75 μg/mL and 50 μg/mL anti-D were incubated with 1e11 amplified phages selected in the first biopanning round for enrichment of high affinity binding. In addition, the bound phages were merely titered but not amplified in the third round. The secondary structure of the synthesised peptide was predicted, based on its amino acid sequence, using PSIPRED (Psi-blast based secondary structure prediction) methods14. The I-TASSER server was employed to predict the three-dimensional structure15. Rasmol, version 2.7.5 (www.RasMol.org) was applied for the visualisation. 3318 IPLLNPGSMQLS(2/26) SPPWLPVMPVRA(1/26) WHYNWQDVSDRQ(3/26) HSYTQTSLWMKV(1/26) 4 Phage display (common panning) 2 PMID:29852986 Recombinant dengue virus type 2 NS1 protein(DENV2 NS1) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) These new peptide-displayed phages or free peptides from phages may have potential applications in dengue diagnosis. 3319 EHDRMHAYYLTR(2/26) 1 Phage display (common panning) 3 PMID:29852986 Recombinant dengue virus type 2 NS1 protein(DENV2 NS1) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) These new peptide-displayed phages or free peptides from phages may have potential applications in dengue diagnosis. 3320 CLNSSQPSC(48/58) CTMSAASHC(5/58) CRGATPMSC(2/58) CFPSHNMHC(1/58) CNHQPSEGC(1/58) CASPFLCPC(1/58) 6 Phage display (in vivo) 6 PMID:29858055 Astrocyte(AS1) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Three rounds against spinal cord in vivo, and then three additional cycles of in vitro panning were performed for each cell line using the phages within 1e9 TU Phage libraries respectively. In total, genomes were isolated from a collection of 50–58 phages. The three cell lines are KT5 cells (AS1; astrocytes), 63 cells (M1; proinflammatory microglia), and Ra2 cells (M2; antiinflammatory microglia). This is the AS1 cell line. 3321 CHHSSSARC(13/51) CNTGSPYEC(3/51) CQTQNRSAC(3/51) CANATCPAC(2/51) CGSKNSAVC(2/51) CNGGTPNRC(2/51) CLKLGEKWC(2/51) CHHRAGVLC(1/51) CHNETQKMC(1/51) CRGATPMSC(1/51) CDLNSDTQC(1/51) CDNPDRRKC(1/51) CDDADQSRC(1/51) CDFPSRGQC(1/51) CDGHDQSLC(1/51) CEHSRPMSC(1/51) CGPMSSKSC(1/51) CLDLPNAMC(1/51) CLNSRQPSC(1/51) CMAPHSRVC(1/51) CMCNSFTWC(1/51) CNDSDTYTC(1/51) CNTDAHRAC(1/51) CQGERWMQC(1/51) CQQSMDPAC(1/51) CSQLPWYSC(1/51) CSDARSPKC(1/51) CTHGLTASC(1/51) CTQSSAMSC(1/51) CTAKGLQAC(1/51) CTCNQMAGC(1/51) CMARYMSAC(0/51) CPNSTHRNC(0/51) CTAHDTNSC(0/51) CDLLHRGAC(0/51) CDFPKLRPC(0/51) CEFSKFRSC(0/51) CGKTSENSC(0/51) CHDLNGSMC(0/51) CIASVDSKC(0/51) CIGNSNTLC(0/51) CLGRTNGQC(0/51) CNRSQQPWC(0/51) CPNMTNQWC(0/51) CQFSKFRSC(0/51) CQRATPGHC(0/51) CRSANIYTC(0/51) CSNSSLTHC(0/51) CSPRPTQTC(0/51) CSWQIGGNC(0/51) CTTINQKVC(0/51) CVPNLQGTC(0/51) CWTDANRDC(0/51) CYTDANRDC(0/51) CYPHNTPNC(0/51) 55 Phage display (in vivo) 6 PMID:29858055 Proinflammatory microglia(MG1) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Three rounds against spinal cord in vivo, and then three additional cycles of in vitro panning were performed for each cell line using the phages within 1e9 TU Phage libraries respectively. In total, genomes were isolated from a collection of 50–58 phages. The three cell lines are KT5 cells (AS1; astrocytes), 63 cells (M1; proinflammatory microglia), and Ra2 cells (M2; antiinflammatory microglia). This is the M1 cell line. 3322 CNTGSPYEC(8/50) CHHSSSARC(7/50) CQTQNRSAC(4/50) CHHRAGVLC(2/50) CMARYMSAC(2/50) CPNSTHRNC(2/50) CTAHDTNSC(2/50) CDLLHRGAC(1/50) CDFPKLRPC(1/50) CEFSKFRSC(1/50) CGKTSENSC(1/50) CHDLNGSMC(1/50) CIASVDSKC(1/50) CIGNSNTLC(1/50) CLGRTNGQC(1/50) CNRSQQPWC(1/50) CPNMTNQWC(1/50) CQFSKFRSC(1/50) CQRATPGHC(1/50) CRSANIYTC(1/50) CSNSSLTHC(1/50) CSPRPTQTC(1/50) CSWQIGGNC(1/50) CTTINQKVC(1/50) CVPNLQGTC(1/50) CWTDANRDC(1/50) CYTDANRDC(1/50) CYPHNTPNC(1/50) CHNETQKMC(1/50) CRGATPMSC(1/50) CDLNSDTQC(0/50) CDNPDRRKC(0/50) CDDADQSRC(0/50) CDFPSRGQC(0/50) CDGHDQSLC(0/50) CEHSRPMSC(0/50) CGPMSSKSC(0/50) CLDLPNAMC(0/50) CLNSRQPSC(0/50) CMAPHSRVC(0/50) CMCNSFTWC(0/50) CNDSDTYTC(0/50) CNTDAHRAC(0/50) CQGERWMQC(0/50) CQQSMDPAC(0/50) CSQLPWYSC(0/50) CSDARSPKC(0/50) CTHGLTASC(0/50) CTQSSAMSC(0/50) CTAKGLQAC(0/50) CTCNQMAGC(0/50) CANATCPAC(0/50) CGSKNSAVC(0/50) CNGGTPNRC(0/50) CLKLGEKWC(0/50) 55 Phage display (in vivo) 6 PMID:29858055 Antiinflammatory microglia(MG2) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Three rounds against spinal cord in vivo, and then three additional cycles of in vitro panning were performed for each cell line using the phages within 1e9 TU Phage libraries respectively. In total, genomes were isolated from a collection of 50–58 phages. The three cell lines are KT5 cells (AS1; astrocytes), 63 cells (M1; proinflammatory microglia), and Ra2 cells (M2; antiinflammatory microglia). This is the M2 cell line. 3323 CHENDVTWC CHENDVTWC CPFSDLNNC CSNFNRYGC CKDVNTSMC 5 Phage display (common panning) 4 PMID:29859290 Acinetobacter baumannii cell Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A. baumannii (A.b.48, A.b.50, A.b.55,A.b.56, and A.b.58) were grown in the medium containing human blood, purified phages (1014 plaque-forming unit (PFU)/ml) were incubated with bacteria grown in BTS (approximately 108 colony-forming unit (CFU)/ml) for 1 h incubation at room temperature (RT). Blood biopanning was done for four rounds. 3324 CERSIRTVC CERSIRTVC CWPMHTRTC CWPMHTRTC CWPMHTRTC CYPHSQESC CRGPTAMSC CNGHTLLSC CERSIRTVC CYLTNTTHC 10 Phage display (common panning) 3 PMID:29859290 Acinetobacter baumannii cell Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Amplified phages (1014 PFU/ml) were incubated with A. baumannii strains (A.b.48, A.b.50, A.b.55, A.b.56, and A.b.58) that formed biofilm, Biofilm biopanning was performed for three rounds. 3325 CRHSQMTVTSRL(15,780) DWSSWVYRDPQT(14,310) SHCLARQTMLQV(9,570) DRWVARDPASIF(7,550) SQDIRTWNGTRS(2,719) VEPPNSQHSGPC(2,467) VHWDFRQWWQPS(2,437) AYYPFVKGRPLP(1,435) MHPNAGHGSLMR(1,428) ASSRRPSVGKGF(1,379) VTTMSGEISQAR(962) GFPTRFEALSSN(828) TGSNSPMWDTEV(722) HVRLTGWQPTWA(306) QFDYMRPANDTH(298) SPCTSFPRCVEL(293) QLATLHKLSGPT(278) 17 Phage display (common panning) 3 PMID:29875202 CD63 protein fragment containing the second extracellular loop (CD63 LEL) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phages (1e12 plaque-forming units) were incubated with CD63 LEL purified protein for 2 hours, and unbound phages were removed by washing with PBS solution for three times. The bound phages were recovered with glycine-HCl buffer (pH 2.2), followed by neutralization with tris-HCl buffer (pH 8.8). Recovered phages were titered and amplified for subsequent cycles. This process was repeated twice, and the recovered phage clones’ sequences were identified by high-throughput sequencing (Genewiz). One peptide in particular, CP05, enabled direct painting of exosomes with different moieties irrespective of the origin of exosomes. 3326 LGISATNAYARH[0.43] FSQATGRSPTTL[0.40] ASVLDYKGFFQR[0.53] TAAQWFPSLSNN[0.26] NWQPTAGLKPLH[0.51] WSAATVPRGFHA[0.76] GALLPSMNKGHW[0.40] SNSDAYALQFLR[0.82] MHTAPGWGYRLS[0.38] FFPLTFPWTYYD[0.51] SGVYKVAYDWQH[0.39] VHWDFRQWWQPS[0.81] YTNPYYSSHTRN[0.75] MNPYPRTPWPHV[0.84] RGMDTLWHHAYH[0.50] 15 Phage display (common panning) 4 PMID:29849110 Folate receptor alpha Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA 94 phage clones were randomLy picked from the third round selection, and their binding afnities for FRα were analyzed individually by phage ELISA at OD450. A Ph.D.-12 phage library displaying random dodecapeptides was used to isolate FRαbinding phages with four rounds of biopanning. Phage yields from each round were used to indicate the enrichment of phages. Afer each round of biopanning, the bound phages were eluted and elutes were tested by phage ELISA to evaluate the afnity to FRα. Peptide sequences of phage clones binding to FRα in random twelve peptide library. 20 clones with the highest ELISA abortion values were sequenced to determine the amino acid sequences of the displayed peptides. 15 clones with individual peptide sequence were identifed. 3327 YDMPNTV[NT] YDMHNTV[NT] YDLHNTV[NT] YDMHNAV[NT] NERALTL[NT] FLSDTRS[0.91] GSGNAIM[1.16] GSGNAHR[NT] 8 Phage display (common panning) 4 PMID:29846456 Anti-Haemonchus contortus polyclonal antibody Haemonchus contortus Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The optical density was determined using a microplate reader (BioTek, USA) at 405 nm at 28 days. Te aim of this study was to evaluate phage display technology for mapping Haemonchus contortus mimotopes. We screened the PhD 7 Phage Display Peptide Library Kit with a sheep polyclonal antibody against H. contortus. After four rounds of selection, 50 phage peptide clones were selected by biopanning and sequenced. 3328 TVNFKLY(20)[0.81] LPQRLRT(5)[0.75] RSLRQRY(2)[0.48] HRRQMPL(2)[0.44] TPKLORM(1)[0.29] 5 Phage display (common panning) 4 PMID:29845357 Anti-M. pneumoniae serum M. pneumoniae Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The ELISA plates were coated with M. pneumoniae-positive serum, negative serum or BSA, respectively. The five representative phages were added and incubated 2 h at 37 °C after blocking. And then HRPlabeled anti-M13 pIII monoclonal antibody was added and incubated for 15 min at 37 °C. The OD450 value was measured. The M. pneumoniae-positive serum was used as the target for biopanning to phage display random 7-peptide library. The representative phages were identifed using dot immunoblotting and ELISA. The exogenous heptapeptides were synthesized and their reactions with M. pneumonia-positive serum were tested by indirect ELISA. Two heptapeptides, namely heptapeptide 1: TVNFKLY and heptapeptide 2: LPQRLRT, were screened out from the randomly selected 40 phages after the four biopanning rounds. Two heptapeptides, namely heptapeptide 1: TVNFKLY and heptapeptide 2: LPQRLRT, were screened out from the randomly selected 40 phages after the four biopanning rounds. 3329 CNTGSPYEC(2.53%)[1.09] CLKLGEKWC(1.10%)[0.87] CSISSLTHC(0.88%)[2.50] CMARYMSAC(0.77%)[2.12] CLMTSQFRC(0.55%)[0.73] CRGATPMSC(0.66%)[0.70] CTVRTSADC[2.47] CNWGDRILC[0.78] CGYSSFNRC[NT] CSHMEYPRC[NT] 10 Phage display (common panning) 3 PMID:29777796 Human breast cancer cell line MDA-MB-231 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The affinity valune were read using an ELISA plate reader at 450 nm. First, MDA-MB-231 cells were prepared as a 1e6 cell suspension in 500 μl of DMEM containing 5% bull serum albumin and were incubated with 1e11 pfu of ph. D.-CX7C phage library on ice for 2 h. Thereafter, unbound phages were washed away. Specific-binding phages were transferred into the organic lower phase and were amplified with coliER2738 host cells. After three rounds, the phages were titrated on plates and were picked up for DNA extraction. As the binding activity of the S3 phage was positively correlated to the level of NKA α1 expression in various breast cancer cells, NKA α1 was validated as the primary target of the S3 peptide. 3330 WHYVRPP(4/24)[1.00] LGSPMSN(4/24)[1.05] NERALTL(4/24)[1.08] FSRPAFL(2/24)[NT] NHSTQMY(2/24)[NT] WHWRYAP(1/24)[NT] VHKTDYL(1/24)[NT] IGNTPAI(1/24)[NT] ANHQSAN(1/24)[NT] EHTRSMN(1/24)[NT] TFASHDQ(1/24)[NT] ITSTFTT(1/24)[NT] HWSTTIS(1/24)[NT] 13 Phage display (common panning) 3 PMID:29770392 Beta-nerve growth factor, Beta-NGF Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The affinity of one clone for each candidate peptide sequence was analyzed with phage ELISA The change in absorbance resulting from color formation was measured by a Spectramax M5 microplate reader (Molecular Devices) at 450 nm wavelength. Then, this value was subtracted from the reference value measured at 650 nm, we take the data at 1.25e9. A high-affinity NGF-binding sequence was identified by phage display and combined with a betasheet forming motif to produce a self-assembling PA molecule. 3331 CDWPDWQCYGWSSHC CKHQAQECVAIREGC CWAMPMWCDSWSQNC CRPQKFNCISANIRC CKAMIGACVAMQFAC CYPVDWYCLFQTVDC 6 Phage display (common panning) 2 PMID:29568446 Bacterial cell wall precursor lipid I Not determined. Not determined. Not determined. CX6CX6C phage display library We identified unique lipid II binding peptides that upon lipidation were found to be active against a range of Grampositive bacteria including clinically relevant strains of vancomycin resistant bacteria. Optimization of the peptide sequence led to variants with enhanced antibacterial activity and reduced hemolytic activity. 3332 CLLQSLLCPYSTHRC 1 Phage display (common panning) 2 PMID:29568446 Enantiomeric Lipid I(ent-Lipid I) Not determined. Not determined. Not determined. CX6CX6C phage display library 3333 AREYGTRFSLIGGYR(7) PKAFQYGGRAVGGLW(7) PVRYGFSGPRLAELW(3) RNVPPIFKEVYWIAQ(3) RTLIRMGTGAHAFAV(3) DRYLPINGVSMFGVP(2) IPVQFSTIDFVAASY(2) PGHSLGKLSVLHSFF(2) QADGPNSVVRPFTLT(2) RSFAYAAAPTSFPWV(2) SVGCPVVGTVGYLRCG(2) VRMFDYGVPRRAVYGG(2) 12 Phage display (in vivo) 5 PMID:29657602 Human breast cancer cell line MCF-7 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) At least 1e12 transducing units of the f3-15mer phage library were intravenously injected into each tumor-bearing mouse through the tail vein and allowed to circulate for 1h before euthanasia by CO2 asphyxiation. Subsequently, 20ml of phosphate-buffered saline buffer was used to wash the unassociated phages by heart perfusion. The tumor-recognized phages were purified from the tumor homogenate and amplified for use as new input phages for the next round of selection. The tumor-homing peptide sequences in every round of selection were identified by phage genome sequencing from the third to fifth round. The subsequent studies primarily focused on the top five peptides with the highest occurrence frequencies, namely, AREYGTRFSLIGGYR (peptide 1), PKAFQYGGRAVGGLW (peptide 2), PVRYGFSGPRLAELW (peptide 3),RNVPPIFKEVYWIAQ (peptide 4) and RTLIRMGTGAHAFAV (peptide 5), because they are more likely to have higher affinity to MCF-7tumors than the other peptides.Using in vivo biopanning, we first selected an MCF-7 breast tumor-targeting peptide, then tested the effectiveness of the as-selected peptide in tumor homing and finally conjugated the peptide to a model photothermal drug, namely, gold nanorods, to achieve enhanced cancer killing efficacy. The peptides identified by the phage display technique can guide the drug to the tumors without the need to know the exact receptors on the tumor. This approach requires significantly less effort to explore patient-specific targeting molecules for precision medicine. 3334 NNHREPPDHRTS GEVGEQEKARVG 2 Phage display (common panning) 3 PMID:29897691 Cryptococcus neoformans H99 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The polypeptide (NNH) has a significant binding to Cryptococcus, which is significantly higher than the other polypeptide (GEV).The experimental results preliminarily showed that the NNH peptide can specifically bind to fungi and will be used as a potential fungal targeting molecule for subsequent research. 3335 VETSQYFRGTLS[0.315±0.045] SSMAPTRSLFVI[0.258±0.026] GSAPLLTVDTSK[0.224±0.022] RYSGLPDIPEPY[0.305±0.042] STTHNPDHSLYR[0.254±0.031] FGSSSTYPLGNW[0.213±0.015] GPIVIYNAPHTN[0.287±0.063] LTGDHSYQIRGA[0.237±0.018] TTNSWHPHNRVL[0.213±0.023] SDGDISLRWYVS[0.276±0.044] ASTPMFSPSHSM[0.229±0.005] SVLEPLVNVTHW[0.202±0.015] SQPWDDSTNRRV[0.272±0.046] VHHASSSFAVHP[0.226±0.038] LPHMGAFNLPLQ[0.195±0.013] NLGDNPLHRDHI[0.273±0.053] GLNNTNYFGVPM[0.226±0.014] SQAQYLHLYPYQ[0.195±0.020] NVSRFLAQPLLP[0.259±0.019] KSIVPSYLDHMQ[0.225±0.009] TTENADLSSRWS[0.192±0.033] 21 Phage display (subtractive panning) 5 PMID:29693344 Human gastric cancer cell line MKN-45 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA After the fifth round of panning, 35 phage clones were randomly selected and sequenced, binding selectivity of each clone was determined by using cellular ELISA with a ratio OD405nm of MKN45 cells to control cells. The culture medium was removed when the HFE-145 cells reached ~70% confluency. 1x1011 pfu phages was added and mixed gently with the blocked HFE-145 cells for 1 h at 37ⅹC. During this time, the MKN-45 cells were pre-washed and blocked in the same manner. Then, the supernatant containing unbound phages was transferred to blocked MKN45 containing wells and incubated for 2 h on a rocker platform at 37℃. The rest of the phage was amplified using E. coli ER2738 host cells for next rounds of biopanning. For each round of panning, 1x1011 pfu of collected phages was used and the detergent (Tween 20) and salt (NaCl) concentrations were increased in a stepwise manner to 0.5%. Peptide VETSQYFRGTLS was selected in the 5th round of biopanning and screened positive as the best binder among all experimentally selected clones. 3336 PFALWQECH GWAVFSDIV GGFLFSDAL LTFVFSDHH GVTVFSDSH TLSPFMEML GGSWFMEWE GVCSFWESG LDWFFYEIV GLTTFVDGA GGAVFVDLG RNAAFLDQH EASIFLESH GFTLFLEDR GMDLFLDTW GGTEFLELV TALLWLELT GMTPFADVC GGFVFADHV GMLLWEEWA GGELFRELY WVTLFHEPH FLILLSEEH GGVPLTELL GGRRLSDDV GGVVLMDSV VCQLLMDHH DLYLLVDDH VYLQLVDEH GQMTLVDLH GVIMLLETI YFVILLDHH GVVVLLDNH AFLLLIEAF VEVLLLEWV YVPLLLDFW GGWFLLDWE WEMLLWDHH RAFVLWDTH GGECLYDAA RVLTLFDWH GGITLFELV VGTGLVRMN RRAELPGGL VVTFFAMDH GGGYYLMLD MLDFFRFHH GVWWYKTWC ELMVYRSHH PEILWRMVH GRRGWRRHH GGGAWRLML GDCEYEGGK GVTLFVWAR GGELFAFAV GGLEWLWGC LLRVWVLSV GGLKWLFTR GCFMWSGSW GGGCFSVWS GLVSFSVYV GGTVFTSLL GTEAFGGGG NVLCFVLGV GATSYVGCV GGGMWGATT AEGLWGSVR GKGLWGPRR 68 Phage display (common panning) 5 PMID:29652924 human cathepsin G (hCG) Leukocyte elastase inhibitor (LEI) Not determined. Not determined. X9 T7 phage display library Each phage clone expresses a unique sequence of 9 random amino acids (nonamer) on their surface, followed by a His6-tag in the C-terminus of capsid protein 10. The phages are subsequently immobilized on Ni-NTA agarose beads via interactions with the His6-tag. The purified hCG was used to screen the phage library for peptides susceptible to cleavage. After the first selection step, the released phages, which are cleaved in their unique random region, were amplified in E. coli and subjected to additional biopannings. Selections of phages susceptible to cleavage by the protease, were performed over 5 biopannings. 3337 CQCWDEADCETGIPC(3)[NT] CLCAEEPDCESLGSC(2)[NT] CGCGGGEDCETGEWC(2)[1.17] CACPQWEDCETGELC(2)[9.52/26.2] CECETGEGCGLGGEC(2)[NT] CGCAGWRDCESGERC(1)[0.36/37.2] CECEAWGDCPSEGLC(1)[NT] CLCEWGLTCPETGEC(1)[8.68] CACPEGRDCETGEPC(1)[NT] CACGGGVECETGERC(1)[NT] CGCWEEEDCETGERC(1)[1.03/2.41] CFCSRESQCEETGEC(1)[NT] CGCRVDEECQKGEPC(1)[NT] CECCAGRDCETGELC(1)[NT] CGCRDDQDCETGEPC(1)[NT] CRCDEELMCVETGEC(1)[NT] CGCWGGADCETGEAC(1)[NT] CRCGGGLDCETGEWC(1)[NT] CECGTGEGCEGWGGC(1)[NT] CPCPLSPGCEETGEC(1)[NT] CGCTEDEECETGEPC(1)[NT] CDCRAVGDCWYAGEC(1)[NT] CGCCWRAVCLYVGGC(1)[NT] [DE]-C-E-[TS]-G-E 23 Phage display (common panning) 3 PMID:29619474 Kelch-like ECH-associated protein 1 Not determined. Not determined. Not determined. CXCX5CX5C M13 phage display library Fluorescence polarization Most of these bicyclic peptides bind with micromolar or sub-micromolar affinities to the target (Ki = 0.36–9.52 mM). One of the two isomers formed by the oxidation of XM-3 exhibits the highest affinity (isomer-I; Ki = 0.36 mM); but the other isomer is a weak binding ligand (isomer-II; Ki = 37.2 mM). The Ki values of the selected peptides were determined using a fluorescence polarization (FP) competition assay as either isomeric mixtures or pure isomers. The Kelchlike ECH-associated protein 1 (Keap1) was used as a target for screening the bicyclic peptide ligands in the library. About 1e13 purified infective phage particles were incubated with the biotinylated target protein captured on magnetic streptavidin or neutravidin beads, and the enriched phages were subjected to two further rounds of selection. To prevent the enrichment of streptavidin and neutravidin binders during panning, the biotinylated protein was captured alternately in each round of selection with streptavidin and neutravidin. 3338 HISSERTHRPROSERSERPRO(7)[25.5] ASPSERSERLEUPHEALALEU(3)[NT] ASNASPLEUMETASNARGALA(2)[3.57] GLYVALALAIIETHRMETLYS(2)[NT] IIEGLYGLYASNLEUSERALA(1)[NT] 5 Phage display (subtractive panning) 3 PMID:29602308 B lymphocyte A20 cell line Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Flow Cytometry Flow cytometry of the binding potential of Peptide-FITC conjugates for target cells. P4-, P6- and P8-FITC conjugates were incubated at 0, 4 or 8 μM with 1e6 A20 cells and analyzed for peptide binding. P4 and P8 peptides demonstrated a strong dose-dependent binding to the target A20 cells. These peptides were then tested for binding specificity by exposure to a series of off-target cells at 4uM, 8uM and 12uM. Clearly, P4 and P8 are taken up by the cells. Ph.D.-7 phage display library was sequentially exposed to a series of normal, control cells in the following order: HUVEC, 3T3, murine heart cell suspension, murine kidney cell suspension, human peripheral blood mononuclear cells, human erythrocytes to remove non-relevant phage clones. After the final absorption, the resultant negatively selected phage pool was termed “adsorbed phage library". A20 cells (1e6) were incubated with the adsorbed phage library for 1 h at 37 °C with gentle shaking. After titration of this lysate, 10–15 isolated plaques were individually recovered, amplified and their DNA extracted according the library manufacturer’s instructions. Peptide inserts were sequenced by Sanger sequencing (Hylabs, Israel) using primers supplied by New England Biolabs and peptide amino acid sequences were deduced. 3339 CTDQTPYVC(2/11)[1.42 ± 0.12] CTDMTPGAC(1/11)[2.24 ± 0.14] CDRTPYRVC(1/11)[0.14 ± 0.03] CFDATPHVC(1/11)[NA] CTDGTPHAC(1/11)[1.41 ± 0.11] CDRTPGRPC(1/11)[0.13] CTDKTPGFC(1/11)[2.10 ± 0.08] CSDRTPYMC(1/11)[0.33] CTDRTPWAC(1/11)[1.30 ± 0.13] CDATPPRLC(1/11)[0.95 ± 0.05] X-D-X-T-P-X-X 10 Phage display (common panning) 4 PMID:29954402 Anti-splicing factor 3b subunit 1(SF3B1) monoclonal antibody, XC24 Splicing factor 3B subunit 1 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Phage ELISA of selected epitopes against XC24 autoantibody were performed. The OD450 values were shown and reproduced from Figure 3b. The specific peptide epitopes against XC24 were selected and, among them, XC24p11 cyclic peptide (CDATPPRLC) was used as an epitope of anti-SF3B1 autoantibody ELISA. With this epitope, we could effectively distinguish between serum samples from hepatocellular carcinoma (HCC) patients (n = 102) and healthy subjects (n = 85) with 73.53% sensitivity and 91.76% specificity (AUC = 0.8731). Moreover, the simultaneous detection of anti-XC24p11 epitope autoantibody and AFP enhanced the efficiency of HCC diagnosis with 87.25% sensitivity and 90.59% specificity (AUC = 0.9081). ELISA using XC24p11 peptide epitope that reacts against anti-SF3B1 autoantibody can be used as a novel test to enhance the diagnostic efficiency of HCC. 3340 SGVYKVAYDCQH SGVYQVAYDWQH ILTCFCPPTWRS SGILTCFCPPRWR 4 Phage display (common panning) 3 PMID:30153280 Anti-glomerular basement membrane (GBM) autoantibody Glomerular basement membrane Not determined. Not determined. Ph.D.-12 phage display library (X12) To identify epitopes recognized by the eluted anti-GBM autoantibody, a combinatorial library peptide (12-mers) phage display kit was utilized (Ph.D.™ 12mer Phage Display Library, New England Biolabs, MA) following manufacturer’s protocol for solution phase panning with affinity bead capture. The bound phages were collected by magnetic field and eluted by incubation for 10 minutes with 1ml of glycine elution buffer (0.2M Glycine-HCl, pH2.2, 1mg/ml BSA). 3341 CGPAEKAYPNNSPLFGPC CGPPPGNPKIKWPPGGPC CGPYIPQPRPPGPRLGPC CGPTGRYTIHTLLRFGPC CGPLFLLRNGYPGKFGPC 5 Bacterial display (common panning) 3 PMID:29961761 Platelet-rich plasma Not determined. Not determined. Not determined. CGPX12GPC bacterial display library (CGPX12GPC) A10 (CGPPPGNPKIKWPPGGPC) was identified from a bacterial peptide display library and synthesized. A10 and protease-activated receptor 4-activating peptide (PAR4-AP, positive control) were immobilized on commercially pure titanium. The peptide-treated titanium showed high epithelial cell migration ability during incubation in platelet-rich plasma. We confirmed the development of dense and expanded BL (stained by Ln5) with pericellular junctions (stained by ZO1) on the peptide-treated titanium surface. In an adhesion assay of epithelial cells on A10-treated titanium, PAR4-AP-treated titanium, bovine root and non-treated titanium, A10-treated titanium and PAR4-AP-treated titanium showed significantly stronger adhesion than non-treated titanium. PAR4-AP-treated titanium showed significantly higher inflammatory cytokine release than non-treated titanium. There was no significant difference in inflammatory cytokine release between A10-treated and non-treated titanium. These results indicated that A10 could induce the adhesion and migration of epithelial cells with low inflammatory cytokine release. This novel peptide has a potentially useful application that could improve clinical outcomes with titanium implants and abutments by reducing or preventing peri-implant disease. 3342 INHQLDTTQILV(5) FPLETSHMSAPL(4) YSSALKTLPIFQ(2) KVFEQDLLTTIL(2) SMQLMTSRLTWN(1) NILTTTWLPLHG(1) TELTTHPVFQFR(1) YQGDANLKTWHV(1) NLVTKPLHVSHL(1) 9 Phage display (subtractive panning) 3 PMID:29990379 CD177 antigen Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) To eliminate peptides that bind to Chinese hamster ovary (CHO) cells, 1e9 phage in PBS containing calcium and magnesium (PBS++), and supplemented with 0.5% bovine serum albumin (BSA), were incubated three rounds with wild-type CHO cells plated on 10 cm tissue culture plates, each for 1 h at room temperature on a rocker. Unbound phage were then incubated with CHO cells expressing human CD177. 3343 DFYKPMPNLRIT(10) WGFKPMDSLVIA(3) DRWVARDPASIF(1) SLDGAGAALRTS(1) GFSHSIPKLVIS(1) SGSTPLFQIFPY(1) 6 Phage display (subtractive panning) 3 PMID:29990379 CD177 antigen Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The library was firstly incubated three rounds with CHO cells expressing HA-FPR to eliminate peptides that bound to the HA-tag. Unbound phage were then incubated with CHO cells expressing mouse CD177. 3344 CTFAGSSC(27.5%) CSASDLSC(9.5%) CNSAFAGC(4.5%) CSYTFAGC(4.5%) CSTFAGSC(4.5%) CRDGYHHC(4.5%) CQNQYPEC(4.5%) CRASAMVC(4.5%) CIDMTHQC(4.5%) CNQSMWSC(4.5%) CQFENGTC(4.5%) CAVKSVTC(4.5%) CNGFMGYC(4.5%) CLTSENAC(4.5%) CRASAMVC(4.5%) 15 Phage display (subtractive panning) 3 PMID:25675522 Purified serum IgG from a year 1 patient with advancing prostate cancer Not determined. Not determined. Not determined. CX6C phage display library In brief, a CX6C (C, cysteine; X, any residue) phage-displayed peptide library was precleared on a pool (50 samples) of protein G-purified antibodies from age-matched healthy donor men to remove nonspecific peptides. Next, the precleared library was incubated with purified serum IgG from a prostate cancer patient of year 1. Three rounds of selection were performed. 3345 CTFAGSSC(60%) CSKKFVTC(15%) CNSAFAGC(5%) CKNKHTTC(5%) CFETFAGC(5%) CNNMYAGC(5%) CQNQYPEC(5%) 7 Phage display (subtractive panning) 3 PMID:25675522 Purified serum IgG from a year 5 patient with advancing prostate cancer Not determined. Not determined. Not determined. CX6C phage display library In brief, a CX6C (C, cysteine; X, any residue) phage-displayed peptide library was precleared on a pool (50 samples) of protein G-purified antibodies from age-matched healthy donor men to remove nonspecific peptides. Next, the precleared library was incubated with purified serum IgG from a prostate cancer patient of year 5. Three rounds of selection were performed. 3346 CTFAGSSC(91%) CNSAFAGC(3%) CFPKRVTC(3%) CPRSAKNC(3%) 4 Phage display (subtractive panning) 3 PMID:25675522 Purified serum IgG from a year 7 patient with advancing prostate cancer Not determined. Not determined. Not determined. CX6C phage display library In brief, a CX6C (C, cysteine; X, any residue) phage-displayed peptide library was precleared on a pool (50 samples) of protein G-purified antibodies from age-matched healthy donor men to remove nonspecific peptides. Next, the precleared library was incubated with purified serum IgG from a prostate cancer patient of year 7. Three rounds of selection were performed. We evaluated binding of the isolated serum antibodies from the different time points to the CTFAGSSC peptide and found an increase in reactivity against the peptide over time (year 0, year 1, year 5 and year 7), with the sample from year 7 producing the most robust binding. Binding of year 7 antibodies to CTFAGSSC could be inhibited in a concentration-dependent manner with preincubation of the synthetic peptide CTFAGSSC, whereas preincubation with a control peptide, CKDRFERC, did not inhibit binding. Immunostaining with autologous purified antibodies on prostate cancer biopsies from the index patient showed reactivity that could be inhibited by preincubation with the synthetic peptide CTFAGSSC. These findings confirm the specificity of the isolated antibodies for the peptide CTFAGSSC. We further identified alpha-2–Heremans–Schmid glycoprotein (AHSG, also known as fetuin-A) as the putative protein corresponding to the peptide mimic. 3347 FHDAWPNLSKSS(8/23) 1 Phage display (common panning) 3 PMID:30123084 Fibroblast growth factor receptor 1, FGFR-1 Fibroblast growth factor 1, FGF-1 1EVT, Not determined. Ph.D.-12 phage display library (X12) ELISA Three positive phage clones with different sequence were used to examine the binding affinity for FGFR1 by ELISA. We found a novel peptide (R1-P2, FHDAWPNLSKSS) with high binding affinity to FGFR1 by using phage display-based screening. We found that peptide R1-P2 inhibited the tyrosine kinase activity of FGFR1 and its downstream ERK/MAPK pathway in A549 and NCI-H460 cells. Peptide R1-P2 also inhibited the proliferation, migration, invasion and promoted the apoptosis of A549 and NCI-H460 cells in vitro. In addition, peptide R1-P2 can effectively suppress tumor growth in xenograft mouse model carrying A549 cells. 3348 VSPPLTLGQLLS(23/44)[2.4 ± 0.01] 1 Phage display (common panning) 3 PMID:23014564 Fibroblast growth factor receptor 3, FGFR-3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA To identify high-affinity FGFR3-binding clones, the recovered phage clones were examined by enzyme-linked immunosorbent assay (ELISA) for FGFR3 binding with control phage(vcsM13). When optical density (OD) values of phage clones were two times greater than that of phage vcsM13, they will be considered to have high binding affinity for FGFR3. The OD450 value was reproduced from Figure 1A. We screened a phage library containing random 12-peptide inserts, using fibroblast growth factor receptor-3 (FGFR3) as bait, and obtained 23 positive clones that share identical amino acid sequences (VSPPLTLGQLLS), named as peptide P3. P3 had high binding affinity to the extracellular domain of FGFR3. We found that P3 inhibited the tyrosine kinase activity of FGFR3 and its downstream ERK/MAPK pathway in chondrocytes. P3 also promoted proliferation and chondrogenic differentiation of cultured ATDC5 chondrogenic cells. In addition,P3 improved the growth of bone rudiments from TDII mice in vitro and rescued the lethal phenotype of mice mimicking human TDII in vivo. 3349 SGQVPWEEPYYVVKKS WEYHYVDLNWTSQHPQ DLPDIIWDFNWETAG 3 Phage display (subtractive panning) 5-6 PMID:30128049 Human sural nerve tissue Not determined. Not determined. Not determined. TriCo-16™ phage display peptide library (X16) Fluorescence To test the affinity of the selected phage display peptides for binding to human nerves, they were chemically synthesized by solid-phase synthesis and labeled with fluorescein Prior to positive selection, phage were counter-selected for high-affinity muscle and fat tissue binding by pre-adsorbing the tissues with the library. For positive selection, phage libraries were mixed directly with human sural nerve tissue. We have now used phage display to identify a novel peptide HNP401 (SGQVPWEEPYYVVKKS) that, when labeled with a fluorophore, selectively binds and highlights human nerves. We show that fluorescently labeled HNP401 can bind to and highlight human sensory and motor nerves such as sural, medial antebrachial cutaneous, laryngeal, ansa cervicalis, great auricular nerve and autonomic nerves like those within and around the prostate gland. 3350 GTGLVRMNH(10) SRLKVHHHH(3) PGGMVAGLG(2) GARTVWGFA(2) FLDGVVRHH(1) PCELVVHHH(1) AVPLVVSGH(1) TPGGVVATY(1) ACADVVLGH(1) GDFRVVLKL(1) GDTTVVVRM(1) MRLGVVVGL(1) TSRPVVVGH(1) GVEEVLGGV(1) GGREVLGKQ(1) RGGAVLVGW(1) GWCLVLSWP(1) GQWGVARAW(1) KEIAVAGQA(1) SMTTVGAKH(1) ERLTVGGHH(1) DRVAVGAHH(1) GTLMVPRTG(1) PGGGVSEVF(1) GAGLVSGGT(1) RARLVSWHH(1) SKAAVSHHH(1) LACNVSHHH(1) LTLCVSHHH(1) TPGGVSCYD(1) TPGGVSSRA(1) GLGAVQTCC(1) GWCQVQSVC(1) GGWEVQVTC(1) GAIRVQDGI(1) TPGGVQLSR(1) TPGGVQKGA(1) RLGLVTGHH(1) ARYPVTLHH(1) TPGGVCINC(1) GFRPVRLAL(1) GGAAVREAV(1) HSVNVRRAH(1) GGRCVRGRS(1) SGPLVHHHH(1) SRASVHHHH(1) PGGLVFDSK(1) GQGPVFLGH(1) LPFLVYKAD(1) LGMMVYGHH(1) ILKHVWHHH(1) GGRTVEWAR(1) GRELVELPC(1) ASCFVDPQH(1) TPGGVDLLC(1) LCRRVDHHH(1) RGASILWTH(1) GCLNILHHH(1) GCSSITGQA(1) GFGLISGVL(1) GWDVISRQR(1) LDMMISSHH(1) RYSPIYMPH(1) GSETIAVGS(1) GSLCIRLRQ(1) QTFFASARH(1) GLQNASAAL(1) TPGGASTAA(1) GYGAASLHH(1) 69 Phage display (common panning) 5 PMID:30459762 N-elastase Not determined. Not determined. Not determined. X9 T7 phage display library An aliquot of the amplified phages were bound to Ni-NTA beads by their His6-tags. Unbound phages were removed by washing ten times in 1.5 ml 1 M NaCl, 0.1% Tween-20 in PBS, pH 7.2, with two subsequent washes with 1.5 ml PBS. The beads were finally resuspended in 1 ml PBS. PBS without protease was used as control. Phages with a random peptide that were susceptible to protease cleavage were released from the Ni-NTA matrix, and the supernatant containing these phages was recovered. To ensure that all of the released phages were recovered the beads were resuspended in 100 μl PBS (pH 7.2) and the supernatant, after mixing and centrifugation, was added to the first supernatant. To ensure that the His6-tags had been hydrolyzed on all phages recovered after protease digestion, 15 μl fresh Ni-NTA agarose beads were added to the combined phage supernatant and the mixture agitated for 15 min followed by centrifugation. A control elution of the phages still bound to the beads, using 100 μl 100 mM imidazole showed that at least 1 × 10^8 phages were attached to the matrix during each selection. 3351 GTLMVPRTG(2) MIWAVSVGF(1) WWVAVSVHH(1) GGIAVSQWL(1) ARWAVSHHH(1) GMCAVSLFD(1) GGVVVSAVW(1) EPLLVSFWH(1) VRLLVSMHH(1) LVLLVSSLM(1) DGFLVSVFL(1) GGVYVSLTI(1) GGLFVSIRW(1) ANFFVSVAL(1) ARIMVSHHH(1) RGLMVSLHH(1) GGRQVSLFW(1) YVTFVMRTH(1) MGIFVMELL(1) GSAFVMLVP(1) WAGEVMLLR(1) GGRTVMLDF(1) ATLDVCHHH(1) VWLVVCVMH(1) AFLAVTFVH(1) SRMLVRLVF(1) EVRLVRMAH(1) YMRAVKGRH(1) IALSVKSML(1) IWSVVHHHH(1) FTLGVHHHH(1) LCTFVEAWY(1) GGLAVEVGL(1) GAMLVELHH(1) GAILVDLLG(1) RFAAVWSVW(1) FGRVVWHHH(1) GGSYVWMIT(1) TPGGVWTAC(1) WLNMVWGVL(1) TPGGVWAWL(1) GARTVWGFA(1) YWWSVWDGW(1) TPGGVFTSW(1) TPGGVFAAT(1) GGFVVFLFN(1) YLDIVYGRH(1) GGMGVYLAF(1) GREVVLRWL(1) YAEVVLLRH(1) AMEDVLHHH(1) LGRFVLHHH(1) GGEFVLWPI(1) GSLFVLYHH(1) GGWFVLAID(1) NWLYVLWDH(1) STWWVLEAH(1) GSRSVLFLC(1) TPGGVLMAW(1) WRWVVAEHH(1) LWAIVAYNW(1) YSTLVAAHH(1) GGLWVAGRW(1) GGVFVALRS(1) GFVDVALTW(1) GGSLVISLL(1) RYFEVGGGH(1) GGMWASDFC(1) GGLFASAAG(1) GGLIARYEW(1) TPGGARFAG(1) AWLDARPWR(1) WGLLANFTL(1) LWLDAGMDT(1) LDIAISLMH(1) KMELIMLHH(1) LRAIIDLGG(1) 77 Phage display (common panning) 5 PMID:30459762 Myeloblastin Not determined. Not determined. Not determined. pComb M13 phage display library An aliquot of the amplified phages were bound to Ni-NTA beads by their His6-tags. Unbound phages were removed by washing ten times in 1.5 ml 1 M NaCl, 0.1% Tween-20 in PBS, pH 7.2, with two subsequent washes with 1.5 ml PBS. The beads were finally resuspended in 1 ml PBS. PBS without protease was used as control. Phages with a random peptide that were susceptible to protease cleavage were released from the Ni-NTA matrix, and the supernatant containing these phages was recovered. To ensure that all of the released phages were recovered the beads were resuspended in 100 μl PBS (pH 7.2) and the supernatant, after mixing and centrifugation, was added to the first supernatant. To ensure that the His6-tags had been hydrolyzed on all phages recovered after protease digestion, 15 μl fresh Ni-NTA agarose beads were added to the combined phage supernatant and the mixture agitated for 15 min followed by centrifugation. A control elution of the phages still bound to the beads, using 100 μl 100 mM imidazole showed that at least 1 × 10^8 phages were attached to the matrix during each selection. 3352 GWAVFSDIV(2) GVCSFWESG(2) TALLWLELT(2) GVIMLLETI(2) PFALWQECH(1) GGFLFSDAL(1) LTFVFSDHH(1) GVTVFSDSH(1) TLSPFMEML(1) GGSWFMEWE(1) LDWFFYEIV(1) GLTTFVDGA(1) GGAVFVDLG(1) RNAAFLDQH(1) EASIFLESH(1) GFTLFLEDR(1) GMDLFLDTW(1) GGTEFLELV(1) GMTPFADVC(1) GGFVFADHV(1) GMLLWEEWA(1) GGELFRELY(1) WVTLFHEPH(1) FLILLSEEH(1) GGVPLTELL(1) GGRRLSDDV(1) GGVVLMDSV(1) VCQLLMDHH(1) DLYLLVDDH(1) VYLQLVDEH(1) GQMTLVDLH(1) YFVILLDHH(1) GVVVLLDNL(1) AFLLLIEAF(1) VEVLLLEWV(1) YVPLLLDFW(1) GGWFLLDWE(1) WEMLLWDHH(1) RAFVLWDTH(1) GGECLYDAA(1) RVLTLFDWH(1) GGITLFELV(1) VGTGLVRMN(1) RRAELPGGL(1) VVTFFAMDH(1) GGGYYLMLD(1) MLDFFRFHH(1) GVWWYKTWC(1) ELMVYRSHH(1) PEILWRMVH(1) GRRGWRRHH(1) GGGAWRLML(1) GDCEYEGGK(1) GVTLFVWAR(1) GGELFAQCL(1) GLELFAFAV(1) GGLEWLWGC(1) LLRVWVLSV(1) GGLKWLFTR(1) GCFMWSGSW(1) GGGCFSVWS(1) GLVSFSVYV(1) GGTVFTSLL(1) GTEAFGGGG(1) NVLCFVLGV(1) GATSYVGCV(1) GGGMWGATT(1) AEGLWGSVR(1) GKGLWGPRR(1) 69 Phage display (common panning) 5 PMID:30459762 Cathepsin G (EC:3.4.21.20) Not determined. Not determined. Not determined. CX10C T7 phage display library An aliquot of the amplified phages were bound to Ni-NTA beads by their His6-tags. Unbound phages were removed by washing ten times in 1.5 ml 1 M NaCl, 0.1% Tween-20 in PBS, pH 7.2, with two subsequent washes with 1.5 ml PBS. The beads were finally resuspended in 1 ml PBS. PBS without protease was used as control. Phages with a random peptide that were susceptible to protease cleavage were released from the Ni-NTA matrix, and the supernatant containing these phages was recovered. To ensure that all of the released phages were recovered the beads were resuspended in 100 μl PBS (pH 7.2) and the supernatant, after mixing and centrifugation, was added to the first supernatant. To ensure that the His6-tags had been hydrolyzed on all phages recovered after protease digestion, 15 μl fresh Ni-NTA agarose beads were added to the combined phage supernatant and the mixture agitated for 15 min followed by centrifugation. A control elution of the phages still bound to the beads, using 100 μl 100 mM imidazole showed that at least 1 × 10^8 phages were attached to the matrix during each selection. 3353 HWWWPAS(6)[0.60] HWTWWNL(1)[0.58] QNTLHPR(1)[0.06] VRIPVWH(1)[0.09] HWWW 4 Phage display (competitive panning) 3 PMID: Lysozyme Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The plate was read at 405 nm using a model M550 microplate reader (BIO-RAD, US). The library phages served as the negative control. ELISA signal values were reprodcued from Figure 4 and shown. The signal of library phages were 0.07. In elution step, the wells were eluted four times and individual elution samples were collected for further assays and/or phage amplification. The elution in each round was conducted as follows: (1) 0.1M glycine–HCl buffer (pH 2.2) containing 1 mg/ml BSA was added in the wells and incubated for 10 min; (2) 200 μl of 0.2 mg/ml lysozyme solution was added and incubated for 60 min; (3) the glycine–HCl buffer was added in the wells again, incubating for 10 min; and (4) the final elution was done by adding 200 μl of 0.2 mg/ml lysozyme solution in the wells and incubating for 120 min. Through comparison of the DNA sequences of foreign peptides of the clones showing specificity to lysozyme molecules, the HWWW motif was found to be the necessary amino acid sequence for the affinity. The electrostatic and hydrophobic interactions are considered to contribute to the affinity for the protein. Moreover, protein chromatography with the immobilized HWWWPAS on Sepharose gel indicated the strong binding affinity of the peptide for lysozyme. 3354 HWWWPAS(15)[1.00] 1 Phage display (competitive panning) 3 PMID: Isulin Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The ELISA signal of the original library phages was used as negative control. It was found that the ELISA signals of the collected phages for insulin reached over 1.0, while that of the control was only 0.164. For the elution of bound phages, the wells were incubated with 0.1 M glycine-HCl buffer (pH 2.2) for 10 min and then with 200 μl of 0.2 mg/mL insulin solution for 60 min. These two elution steps were repeated and the phages collected in the fourth elution were amplified for the next round biopanning and screening. The peptide (HWWWPAS) was synthesized and coupled to EAH Sepharose 4B (5.4 μmol/mL bed). Then, insulin chromatography was carried out with mobile phases of different pH values and by the addition of urea and ethylene glycol. It was found that electrostatic interactions were predominant for the affinity binding, and hydrogen bonding might also contribute somewhat to the affinity. Finally, frontal analysis was performed and the dynamic binding capacity of the affinity column for insulin at 50% breakthrough was estimated at 60.6 mg/mL bed, which was about two times higher than the theoretical binding capacity of the monomeric insulin. The result suggests that insulin was bound in dimer state in a stoichiometric relationship with the coupled peptide, indicating the high binding efficiency of the peptide ligand for insulin. 3355 GGVCFRVGLSIVCNF ALVFFFSSGDFYGVS GSCRAFLSGVVCSFP 3 Phage display (common panning) 4 PMID:15377604 Lactotransferrin (EC:3.4.21.-), Lactoferrin Not determined. Not determined. Not determined. f3-15mer phage display library (X15) The custom synthesized peptide HuH6 (GSCRAFLSGVVCSFP) displayed on the HuLF binding phage isolated from the linear pentadecamer library, however, did not bind lactoferrin, as detected with Surface Plasmon Resonance, nor did it inhibit the phage binding to lactoferrin. 3356 CAPRQPPC CRTSLKVC CDTNREQC CDQDQDTC CDQVAKGC CEKGQRRC CEGKQRRC CTKGNQPC CTRSGTRC CSATEHGC CPGGYTKC CTDNAKEC CDVGQNKC CHQHRQRC 14 Phage display (common panning) 2 PMID:15377604 Lactotransferrin (EC:3.4.21.-), Lactoferrin Not determined. Not determined. Not determined. CX6C phage display library The peptides HuN (CEGKQRRC) and HuL (CDQVAKGC) selected from the cyclic hexamer library, custom synthesized did neither bind lactoferrin nor inhibit the phage binding to lactoferrin. Furthermore, reducing both peptides to obtain a linear form did not change this. 3357 CIPASKAC CGLTGLAC CDRPNAKC CPKHRNKC CPRSQQEC CQWLQHPC CPYTYWTC CTAPRRAC CRKSVHSC CSKVKNRC CSFRGQSC CYQSQNGC CHGKSANC CGKFSYQC CPKTERRC 15 Phage display (common panning) 2 PMID:15377604 Bovine lactoferrin Not determined. Not determined. Not determined. CX6C phage display library 3358 TRWLVYFSRPYLVAT(8)[4.4] PRHVFYRWFLSNPRI(4)[NT] IVRHLFLHVYPRVLM(1)[NT] 3 Phage display (common panning) 4 PMID: Barley α-amylase 1 Not determined. Not determined. Not determined. f3-15mer phage display library (X15) ELISA The phage clone was analyzed by measuring the dissociation constant using an indirect competition method based on ELISA. All three peptides share a common feature of an Arg residue flanked by high numbers of Tyr and Trp/Phe residues. The phage display peptide TRWLVYFSRPYLVAT showed an activity 5-fold greater than those of the other two peptides and a dissociation constant of 4.4 nM. 3359 RLRSFY HLNSLS RVLSAH KGTTYY EINVPT SFAKLS LPPPKL SHLPWH 8 Phage display (common panning) 3 PMID:8804536 von Willebrand factor, vWF Type I collagen Not determined. Not determined. f3-6mer phage display library (X6) Phages from the 6-mer bacteriophage display library were added to von Willebrand Factor (vWF). The solution was then transferred into the recombinant Factor VIII (rFVIII) coated microtiter wells to effect capture of vWF and vWF:phage complexes by recombinant Factor VIII, which was designed to avoid selection of peptides binding to vWF at FVIII contact sites. Phage peptide library screening yielded a lead peptide (RLRSFY) that interacts with vWF. Conservative substitutions of terminal residues of the lead peptide led to a second peptide, RVRSFY, which was more efficient in the affinity chromatographic purification of vWF from protein mixtures. Adsorption isotherm measurements indicated multiple interactions between vWF and the immobilized peptide RVRSFY. Increases in peptide density on the chromatographic supports resulted in stronger association constants and higher maximum protein binding capacities. When the peptide density was lower than 32 mg/mL, there was no measurable interaction between vWF and immobilized peptide RVRSFY in HEPES buffer containing 0.5 M NaCI at pH 7. An increase in peptide density from 32 to 60 mg/mL increased the association constants from 0.9e6 to 2e6 (1/M). Divalent salts (calcium and magnesium chloride) were used to elute the retained vWF with 82.5% of the activity recovered. The interactions between vWF and the immobilized peptide RVRSFY are dominated by ionic attractions and also involve hydrophobic interactions at close contact. Finally, the purification of vWF from crude material PEG filtrate of a cryoprecipitate of human plasma is demonstrated using affinity chromatography with immobilized N-acetyl-RVRSFYK. 3360 KHRFNKD(2)[1.62 ± 0.23] PIISRAT(2)[0.15 ± 0.02] WSMRGTT(1)[NT] PGPSTTS(1)[NT] SKPLHKM(1)[NT] HRRPSRS(1)[NT] QISSLRA(1)[NT] WTDPHPP(1)[NT] HLAPAQY(1)[NT] 9 Phage display (competitive panning) 4 PMID: Anti-goat immunoglobulin G monoclonal antibody Goat immunoglobulin G Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The immunoplate and absorbance was estimated at 405 nm using a Synergy-HT microplate reader (Biotek, Winooski, USA). Each value was obtained by subtracting the value for the control from the absorbance value corresponding to each peptide. The control absorbance was determined using an immunoplate treated with blocking buffer only. The absorbance value was reproduced from Figure 2 and shown. Goat immunoglobulin G (IgG, 150 μg) was added to a culture dish with 1.5 mL TBST (0.1% Tween-20, 150 mM NaCl, 50 mM Tris-Cl, pH 7.5). The dish was incubated overnight at 4℃ and washed with TBST. Then, the dish was filled with blocking buffer (1 mM EDTA, 0.25% BSA in TBST), incubated at room temperature for 1 h, and washed with TBST. Rabbit anti-goat IgG antibody (27 μg) was added to the dish with 1 mL TBS (150 mM NaCl, 50 mM Tris-Cl, pH 7.5) and allowed to react at room temperature for 1 h. After washing, phages (2e11 pfu diluted in 1 mL TBS) were added to the dish and allowed to react at room temperature for 1 h. Rabbit anti-goat IgG antibody (27 μg in 1 mL TBS) was added to the dish and allowed to react at room temperature for 1 h for competitive elution of phages specific to rabbit antibody. One of these sequences (KHRFNKD), which was rich in positive charge and homologous to the IgG-binding domains of Staphylococcal protein A, exhibited a high and specific affinity to rabbit antibody. This peptide was immobilized on a quartz crystal microbalance (QCM) biosensor, and the resulting QCM was found to be capable of capturing antibody and detecting antigen, with potential applications in antibody purification and enzyme immunoassay. 3361 CPLSPSHEC(5)[0.08 ± 0.03] CAAPSPRNC(1)[NT] CPRTEKRRC(1)[NT] CQRSGKPSC(1)[NT] CPAGADWPC(1)[NT] CEPTSHFHC(1)[NT] CSSGSDYTC(1)[NT] 7 Phage display (competitive panning) 4 PMID: Anti-goat immunoglobulin G monoclonal antibody Goat immunoglobulin G Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The immunoplate and absorbance was estimated at 405 nm using a Synergy-HT microplate reader (Biotek, Winooski, USA). Each value was obtained by subtracting the value for the control from the absorbance value corresponding to each peptide. The control absorbance was determined using an immunoplate treated with blocking buffer only. The absorbance value was reproduced from Figure 2 and shown. Goat immunoglobulin G (IgG, 150 μg) was added to a culture dish with 1.5 mL TBST (0.1% Tween-20, 150 mM NaCl, 50 mM Tris-Cl, pH 7.5). The dish was incubated overnight at 4℃ and washed with TBST. Then, the dish was filled with blocking buffer (1 mM EDTA, 0.25% BSA in TBST), incubated at room temperature for 1 h, and washed with TBST. Rabbit anti-goat IgG antibody (27 μg) was added to the dish with 1 mL TBS (150 mM NaCl, 50 mM Tris-Cl, pH 7.5) and allowed to react at room temperature for 1 h. After washing, phages (2e11 pfu diluted in 1 mL TBS) were added to the dish and allowed to react at room temperature for 1 h. Rabbit anti-goat IgG antibody (27 μg in 1 mL TBS) was added to the dish and allowed to react at room temperature for 1 h for competitive elution of phages specific to rabbit antibody. 3362 SAGHLTHGGGWAVPFYSHSTPARYAN YFPKWKVGGGSAVPFYSHSMESASAL SPLIWNWGGGSAVPFYSHSHMKSQSP DVNKYYLGGGSAVPFYSHSSRSEDYN HPFLRLPGGGSAVPFYSHSFQSPMLP 5 Phage display (competitive panning) 3 PMID:11397457 Lipase (EC:3.1.1.3) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Biopanning was performed in microtiter plate wells. Elution of the phage was carried out using 100 μl of 100 μg/ml lipase solution in TBST. 3363 HYSPFRTGGGSAVPFYSHSPSTFMSR 1 Phage display (competitive panning) 6 PMID:11397457 Lipase (EC:3.1.1.3) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Biopanning was performed in microtiter plate wells. Elution of the phage was carried out using 100 μl of 100 μg/ml lipase solution in TBST. 3364 SSRLMIL AAWLSPL GSFLYHP SFTHLLY LTPVYYW SSHMPHW 6 Phage display (competitive panning) 3 PMID:11397457 Lipase (EC:3.1.1.3) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Biopanning was performed against lipase attached to magnetic beads. Bound phage were eluted by resuspending the beads in 100 μl of 100 μg/ml lipase solution in TBST, incubated at room temperature for 1 h. The synthetic heptapeptide GSFLYHP did not act as a satisfactory affinity ligand after immobilization on Sepharose. This indicated that the interaction with lipase was due to both the heptapeptide and the presence of a part of the phage coat protein. This conclusion was further verified by immobilizing the whole phage on the surface of magnetic beads and using the resulting conjugate as an affinity adsorbent. 3365 GETRAPL(3)[n.a.] TVRTSAD(2)[n.a.] YPSPWGY(2)[n.a.] ISLPSPT(1)[0.25 ± 0.02] SSQLSRP(1)[0.24 ± 0.01] YNSPTHH(1)[0.30 ± 0.03] APKQSLE(1)[n.a.] APTAVSK(1)[n.a.] ETTHLTG(1)[n.a.] GGLSSRP(1)[n.a.] GVPTALP(1)[n.a.] KPFPPMK(1)[n.a.] LGTSMQL(1)[n.a.] LPLWEVY(1)[n.a.] LTAQPST(1)[n.a.] MISSNSS(1)[n.a.] QLKPLEF(1)[n.a.] QLVYPAP(1)[n.a.] SGTHHKA(1)[n.a.] SLGPSPG(1)[n.a.] SPHDVAYD(1)[n.a.] TLNRVPN(1)[n.a.] TNTMTRA(1)[n.a.] TPHPLRL(1)[n.a.] VNRSSLY(1)[n.a.] VSTPSTP(1)[n.a.] 26 Phage display (common panning) 4 PMID:30268850 Cystic fibrosis (CF) mucus Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA To understand binding affinities between phage-displayed peptides and the major protein component of mucus (mucin), ELISA was performed. A mucin solution was used instead of CF mucus since other mucus components such as lecithin or BSA might block the binding of mucin protein to the wells. Phage clones SSQ-phage, ISL-phage, and YNS-phage binding to mucin was measured by ELISA and compared to wild-type phage. The anti-M13 antibody used for detection binds specifically to an epitope on the pVIII major coat protein. Thus the binding affinities measured correlate with phage-mucin interactions. M13KE wild-type phage demonstrated a statistically significant higher binding affinity for mucin than phage clones SSQ-phage, ISL-phage, and YNS-phage Mucin only and monoclonal antibody only controls did not show specific binding, confirming that the measured binding affinities are specific to interactions between phage and mucin. The affinity value was measured at 450nm. The absorbance (450nm) value was reproduced from Figure 5 and shown. An initial concentration of 2e10 phage pfu was mixed with cystic fibrosis (CF) mucus, and 250 μL of the mixture was transferred to the apical compartment (donor) in the transwell system containing 1.5 mL phosphate buffered saline (PBS) in the basolateral compartment (receiver) and incubated for 1 h at room temperature (25°C). After 1 h, the eluate was collected from the basolateral side and titered using standard double-layer plaque assay to quantify phage concentration. The eluted phage library was amplified in XL-1 Blue E. coli (Agilent, CA) to make more copies, which were quantified by plaque assay prior to the next round of screening. Equivalent amounts of input phage were added for each round. This iterative screening was performed for a total of four rounds. Here, phage clones displaying discovered peptides were shown to have up to 2.6-fold enhanced diffusivity in the CF mucus model. In addition, we demonstrate reduced binding affinities to mucin compared to wild-type control. 3366 GCTRMICN(2) GCTRMLCR(2) GCTRMMCR(2) ACTKMMCQ(1) ACTRMACA(1) ACTRMRCS(1) GCTKKMCG(1) GCTKMICR(1) GCTKMLCG(1) GCTKMLCL(1) GCTKMMCG(1) GCTKQLCV(1) GCTRIMCG(1) GCTRMGCV(1) GCTRMICG(1) GCTRMLCG(1) GCTRMLCN(1) GCTRMLCT(1) GCTRMLCS(1) GCTRMMCA(1) GCTRMMCE(1) GCTRMMCS(1) GCTRMQCS(1) GCTRMVCG(1) GCTRMVCM(1) GCSRMICE(1) GCSRMLCD(1) GCSRMLCG(1) GCSRMLCV(1) GCSRMMCG(1) GCSRMRCD(1) LCTKMYCG(1) MCTKMLCK(1) MCTRMFCM(1) MCTRMMCR(1) MCTRMSCF(1) NCTRMMCN(1) QCTRMFCS(1) SCTRMACQ(1) SCTRMACR(1) SCTRMHCG(1) SCTRMLCG(1) SCTRMRCN(1) SCTRMSCG(1) SCSRMLCF(1) YCTRMMCE(1) 46 Phage display (common panning) 3 PMID:30543823 Trypsin Pancreatic trypsin inhibitor Not determined. Not determined. SGPI-2 XCX4CX M13 phage display library Briefly, bovine trypsin was immobilized on MaxiSorp (Nunc) plates. Phage particles were obtained with the PEG/NaCl precipitation method and applied to the blocked wells. Following thorough washing, bound phages were eluted. E. coli XL1-Blue was used to amplify the eluted phages to be used in a next round of selection. After three selection and amplification cycles, the libraries were enriched over 100-fold compared to the control obtained on blocked wells containing no target enzymes. 3367 GCARHFAP(1) GCARIYSP(1) GCARNFDP(1) GCDKASRF(1) GCDRRLSF(1) GCDRSLNY(1) GCDRSVMY(1) GCIKLYSP(1) GCNKGLWM(1) GCNKIVHP(1) GCNRILDW(1) GCPRFLDF(1) GCPRGLSF(1) GCPRHLVP(1) GCPRHYIP(1) GCPRHYYP(1) GCPRIIYP(1) GCPRILAS(1) GCPRILDY(1) GCPRILQP(1) GCPRILSL(1) GCPRIMAP(1) GCPRIYFP(1) GCPRLFEF(1) GCPRLFHF(1) GCPRLLSP(1) GCPRLYLP(1) GCPRMVDA(1) GCPRSLDF(1) GCPRVHFP(1) GCPRVINP(1) GCPRVLDT(1) GCPRVVDL(1) GCPRYLSL(1) GCRRNIDP(1) GCTKIYDP(1) GCTRDLRL(1) GCTRELAP(1) GCTRIFSP(1) GCTRSLDF(1) GCSKQLSF(1) GCSRALNF(1) GCSRHLHY(1) GCSRHLNP(1) GCSRHLSP(1) GCSRIFNP(1) GCSRIHAP(1) GCSRILNP(1) GCSRMLDP(1) GCSRNFEP(1) GCSRNYAP(1) GCSRQLDL(1) GCSRVLSF(1) GCSRVRDD(1) GCSRVYAP(1) GCSRVYSP(1) 56 Phage display (common panning) 3 PMID:30543823 Trypsin Pancreatic trypsin inhibitor Not determined. Not determined. SPINK1 GCX6 M13 phage display library Briefly, bovine trypsin was immobilized on MaxiSorp (Nunc) plates. Phage particles were obtained with the PEG/NaCl precipitation method and applied to the blocked wells. Following thorough washing, bound phages were eluted. E. coli XL1-Blue was used to amplify the eluted phages to be used in a next round of selection. After three selection and amplification cycles, the libraries were enriched over 100-fold compared to the control obtained on blocked wells containing no target enzymes. 3368 ACTLKMCK(1) ACTLRACI(1) ACTMKMCR(1) ACTMMMCK(1) ACTMRACP(1) ACTYRRCA(1) DCTFKLCR(1) DCTMKFCQ(1) DCTMRRCK(1) DCTQKACR(1) DCTWKLCR(1) DCTWKRCT(1) DCTWRLCN(1) DCTWRMCR(1) DCTWRRCR(1) GCSWMRCS(1) GCTFMRCQ(1) GCTLKMCR(1) GCTLRLCA(1) GCTLRRCR(1) GCTMIRCK(1) GCTMKLCG(1) GCTMKLCR(1) GCTMKRCK(1) GCTMMLCR(1) GCTMMMCK(1) GCTMRACR(1) GCTMRMCQ(1) GCTMRRCR(1) GCTMVMCR(1) GCTMVRCK(1) GCTWKKCR(1) LCTYRMCR(1) MCTMMWCR(1) MCTWKMCA(1) MCTWKMCK(1) SCSWKRCN(1) SCSWKRCR(1) SCTLKLCM(1) SCTLKRCK(1) SCTMKLCK(1) SCTMKMCK(1) SCTMKRCR(1) SCTMRRCK(1) SCTMRRCR(1) SCTWRLCK(1) TCTLKRCR(1) TCTMKMCR(1) VCTMKMCR(1) VCTWKMCG(1) VCTWKMCR(1) VCTYALCR(1) 52 Phage display (common panning) 3 PMID:30543823 Chymotrypsin Not determined. Not determined. Not determined. SGPI-2 XCX4CX M13 phage display library Briefly, chymotrypsin was immobilized on MaxiSorp (Nunc) plates. Phage particles were obtained with the PEG/NaCl precipitation method and applied to the blocked wells. Following thorough washing, bound phages were eluted. E. coli XL1-Blue was used to amplify the eluted phages to be used in a next round of selection. After three selection and amplification cycles, the libraries were enriched over 100-fold compared to the control obtained on blocked wells containing no target enzymes. 3369 GCAFVMKP(1) GCPYMIRP(1) GCAYVLRP(1) GCTLVFRP(1) GCAFELRP(1) GCSFVLKP(1) GCNFKLSP(1) GCTLMYRP(1) GCPLILKP(1) GCSFIMRP(1) GCAFTLRP(1) GCPYKLSW(1) GCPFLLRF(1) GCTYLHRP(1) GCSWIYKP(1) GCTWLYRP(1) GCPFALRL(1) GCSFQLRQ(1) GCLFQLRP(1) GCSFELRP(1) GCSWVLSP(1) GCSFILRP(1) GCTYLYRP(1) GCTFVLKP(1) GCNFMFRP(1) GCAYLLRP(1) GCHFILKP(1) GCTFILRP(1) GCTFVYSP(1) GCSWQLRP(1) GCTFLLRP(1) GCVLSLRP(1) GCTFYLKP(1) GCVYIFRP(1) GCTMILKP(1) GCTLQLRP(1) GCAMILRP(1) GCGWILRP(1) GCSWVLRP(1) GCAYTLRP(1) GCTMVLRP(1) GCNYILKP(1) GCQWSLRP(1) GCTMEWRP(1) GCSMIFKP(1) GCTFVLRP(1) GCFYLLRP(1) GCPFILKL(1) GCPLIFKP(1) GCTYILRP(1) GCSFLLRP(1) GCPYVLKP(1) GCALMLRP(1) GCAFVIRP(1) GCTFSLRA(1) GCAFSLRR(1) 56 Phage display (common panning) 3 PMID:30543823 Chymotrypsin Not determined. Not determined. Not determined. SPINK1 GCX6 M13 phage display library Briefly, chymotrypsin was immobilized on MaxiSorp (Nunc) plates. Phage particles were obtained with the PEG/NaCl precipitation method and applied to the blocked wells. Following thorough washing, bound phages were eluted. E. coli XL1-Blue was used to amplify the eluted phages to be used in a next round of selection. After three selection and amplification cycles, the libraries were enriched over 100-fold compared to the control obtained on blocked wells containing no target enzymes. 3370 KKQKCRTDACVTQM(7)[128.6 ± 23.5] NEKKCKGARCTTVT(3)[84.3 ± 19.4] ATPKCKKKSCMTTQ(3)[94.5 ± 29.9] VDKKCKSDDCGAWH(3)[n.a.] ETKKCTTGPCKVVT(2)[13.4 ± 4.7] HDKKCKRQPCVLAN(2)[n.a.] FDKKCKSNKCLEVR(2)[n.a.] PKKKCHPEPCQTCG(2)[n.a.] KTEHCKKRKCPLDM(2)[n.a.] KKKKCKKKICTTHT(1)[n.a.] KKKKCKKNTCKNHT(1)[n.a.] 11 Phage display (common panning) 3 PMID:29928986 Terbium doped cerium-magnesium aluminate (CAT) Not determined. Not determined. Not determined. F88-FUSE X4CX4CX4 phage display library (X4CX4CX4) Binding assay Pre-identified phage, derived from single clone amplification out of colonies of infected cells, were characterized concerning their ability to bind to individual components of fluorescent lamp powder and to quantify the selective binding behavior. Components tested were CeMgAl11O19:Tb3þ (CAT), Y2O3:Eu3þ (YOX), BaMgAl10O17:Eu2þ (BAM), LaPO4:Ce3þ,Tb3þ (LAP) and halophosphate (HP). Mineral preparation, phage production and binding studies were performed. Mineral dispersions were incubated together with the phage and subsequently repeatedly washed, resulting in the removal of weakly bound phage. Phage concentrations in input and eluate were determined and compared to the binding behavior of wild-type fd-tet phage (obtained by Dr. Jamie Scott). Binding efficiency was calculated using phage titer before and after binding assays in comparison to the wild-type phage. Random peptide sequences with high affinity to CAT minerals were identified from the PVIII phage library Cys4. After three rounds of biopanning corresponding DNA sequences of the displayed peptides on phage of 80 infected colonies were amplified via PCR and sequenced. 11 different sequences were determined. 3371 CMSTGLSSC(28)[12.8 ± 0.7] CVPILEGTC(5)[21.5 ± 0.3] CDRTISNKC(2)[31.8 ± 0.5] CQNPGNTLC(2)[2.5 ± 0.9] CSGTGASYC(2)[1.7 ± 0.7] CSSSVVTHC(2)[2.3 ± 0.9] CDAKDLNSC(1)[2.5 ± 0.9] CDNDTKASC(1)[1.5 ± 0.5] CGLTDTSNC(1)[0.8 ± 0.3] CKTSTHAIC(1)[1.5 ± 0.3] CMRDSKMLC(1)[2.2 ± 0.6] CSTISKAKC(1)[2.6 ± 0.8] CTASQNFYC(1)[1.4 ± 0.2] CTGQGGEYC(1)[1.5 ± 0.2] CTKTQTHAC(1)[0.8 ± 0.0] CTNHSAYHC(1)[2.7 ± 2.0] CTQMLGQLC(1)[1.4 ± 0.27] CVSPNKEAC(1)[3.1 ± 0.7] 18 Phage display (common panning) 3 PMID:29928986 Serum 13006 from patients sensitized to soy bean Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Binding assay Pre-identified phage, derived from single clone amplification out of colonies of infected cells, were characterized concerning their ability to bind to individual components of fluorescent lamp powder and to quantify the selective binding behavior. Components tested were CeMgAl11O19:Tb3þ (CAT), Y2O3:Eu3þ (YOX), BaMgAl10O17:Eu2þ (BAM), LaPO4:Ce3þ,Tb3þ (LAP) and halophosphate (HP). Mineral preparation, phage production and binding studies were performed. Mineral dispersions were incubated together with the phage and subsequently repeatedly washed, resulting in the removal of weakly bound phage. Phage concentrations in input and eluate were determined and compared to the binding behavior of wild-type fd-tet phage (obtained by Dr. Jamie Scott). Binding efficiency was calculated using phage titer before and after binding assays in comparison to the wild-type phage. Biopanning was carried out on planar sol–gel surfaces. Sol-gels with cation exchange properties were fabricated using Dispex N40, polyacrylic acid or polystyrene sulfonate.63 colonies were examined for their peptide sequence. 3372 CSGTGASYC(3)[5.9 ± 1.7] CMSTGLSSC(2)[4.0 ± 2.9] CNTGSPYEC(2)[1.2 ± 0.0] CTASQNFYC(2)[10.4 ± 0.0] CGSRSAQTC(1)[2.0 ± 1.1] CGTKGSLNC(1)[17.8 ± 3.0] CGYSSFNRC(1)[3.6 ± 1.2] CHHPVANTC(1)[1.1 ± 0.2] CHNETQKMC(1)[1.6 ± 0.6] CKDTSRSAC(1)[1.2 ± 0.1] CNAKHHPRC(1)[16.8 ± 6.5] CNGRAVNYC(1)[0.9 ± 0.5] CPGASVTYC(1)[1.1 ± 1.0] CRAEGTSEC(1)[1.0 ± 0.0] CRGATPMSC(1)[1.0 ± 0.7] CSLATDQKC(1)[2.1 ± 0.8] CSNNHSSMC(1)[2.2 ± 1.0] CSTATPYKC(1)[2.4 ± 1.5] CTKTDVHFC(1)[2.6 ± 2.0] CTSVLNNTC(1)[2.4 ± 0.4] CVPILEGTC(1)[1.9 ± 0.8] 21 Phage display (common panning) 3 PMID:29928986 Nickel ion Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Binding assay Pre-identified phage, derived from single clone amplification out of colonies of infected cells, were characterized concerning their ability to bind to individual components of fluorescent lamp powder and to quantify the selective binding behavior. Components tested were CeMgAl11O19:Tb3þ (CAT), Y2O3:Eu3þ (YOX), BaMgAl10O17:Eu2þ (BAM), LaPO4:Ce3þ,Tb3þ (LAP) and halophosphate (HP). Mineral preparation, phage production and binding studies were performed. Mineral dispersions were incubated together with the phage and subsequently repeatedly washed, resulting in the removal of weakly bound phage. Phage concentrations in input and eluate were determined and compared to the binding behavior of wild-type fd-tet phage (obtained by Dr. Jamie Scott). Binding efficiency was calculated using phage titer before and after binding assays in comparison to the wild-type phage. Biopanning was carried out on planar sol–gel surfaces. Sol-gels with cation exchange properties were fabricated using Dispex N40, polyacrylic acid or polystyrene sulfonate.29 colonies were examined for their peptide sequence. 3373 KVWILPA(6/30) KVWVIPS(4/30) 2 Phage display (common panning) 3 PMID:30521966 Virus-like particle (VLP) of orange-spotted grouper nervous necrosis virus (OGNNV) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Briefly, 150 μL of the purified RBS (100 μg/mL) were coated with coating buffer (0.1 M NaHCO3, pH 8.6) in each well of the 96-well plate. After blocking with BSA (5 mg/mL) in coating buffer and washing 5 times with 0.1% TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% [v/v] Tween-20, pH 7.5), the well containing RBS was incubated with 100 μL of diluted library (5 × 1011 sequences) for 1 h at room temperature. After washed 10 times with TBST, bound phages were eluted with elution buffer (0.2 M glycine, 1 mg/mL BSA, pH 2.2) and neutralized with 1 M Tris (pH 9.1). Eluates were incubated with Escherichia coli ER2738 cells and amplified for 5 h at 37 °C. The amplified phages were collected, purified, and titrated for the next round of surface panning procedure. 3374 VHWDFRQWWQPS(11/30) 1 Phage display (common panning) 3 PMID:30521966 Virus-like particle (VLP) of orange-spotted grouper nervous necrosis virus (OGNNV) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Briefly, 150 μL of the purified RBS (100 μg/mL) were coated with coating buffer (0.1 M NaHCO3, pH 8.6) in each well of the 96-well plate. After blocking with BSA (5 mg/mL) in coating buffer and washing 5 times with 0.1% TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% [v/v] Tween-20, pH 7.5), the well containing RBS was incubated with 100 μL of diluted library (5 × 1011 sequences) for 1 h at room temperature. After washed 10 times with TBST, bound phages were eluted with elution buffer (0.2 M glycine, 1 mg/mL BSA, pH 2.2) and neutralized with 1 M Tris (pH 9.1). Eluates were incubated with Escherichia coli ER2738 cells and amplified for 5 h at 37 °C. The amplified phages were collected, purified, and titrated for the next round of surface panning procedure. From the positive clones, a dodecapeptide (VHWDFRQWWQPS) with the highest binding capacity (BC) to RBS (virus-like particle (VLP) of orange-spotted grouper nervous necrosis virus (OGNNV)) was selected. This affinity peptide (AFP) agglutinated or disrupted virion particles, inhibiting RBS entry into sea bass (SB) cells. To enhance BC and solubility, we amended the AFP sequence as “LHWDFQSWVPLL” and named as 12C. One to three copies of 12C in tandem were prokaryotically expressed with a maltose binding protein (MBP) linked by a flexible peptide. Of the recombinant proteins expressed, MBP-triple-12C (MBP-T12C) exhibited the highest BC, efficiently blocked RBS entry, and strongly inhibited OGNNV infection at viral entry. Moreover, MBP-T12C bound the VLPs of all nervous necrosis virus (NNV) serotypes, displaying broad-spectrum anti-NNV ability, and recognized only OGNNV and mud crab virus, demonstrating binding specificity. Therefore, these anti-NNV AFPs specifically bound NNV, aggregating or disrupting the viral particles, to reduce the contact probability between the virus and cell surface, subsequently inhibiting viral infection. Our results not only provided a candidate of anti-NNV AFP, but a framework for the development of antiviral AFP. 3375 CDNVAQSVC(6)[4059.0 ± 213.5] CKTSPWAKC(1)[n.a.] 2 Phage display (common panning) 3 PMID:30431079 C57BL/6 mouse bone marrow mesenchymal stem cell (BMMSC) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Flow Cytometry The identified peptide (CDNVAQSVC) was designated as a mouse BMMSCs affinity peptide (D7). A peptide (CVAVQNDSC) with the same amino acids as D7 in a scrambled order was used as the negative control and termed V7. RGD, a peptide composed of 3 amino acids (arginine, glycine and aspartic acid), was used as the positive control. The C57BL/6 mouse BMMSCs were incubated for 1 h with FITC-labeled D7, V7 or RGD, and measured via flow cytometry. The average fluorescence intensity was 3,058.5 ± 604.6 for the mouse BMMSCs incubated with FITC-RGD, and 146.5 ± 60.1 for the mouse BMMSCs incubated with FITC-V7. The average fluorescence intensity was shown for the mouse BMMSCs incubated with FITC-D7. The phage display library consisted of e9 different phages. Cells plated in 60x15 mm petri dishes were rinsed twice with PBS (Biological Industries, Ltd.; cat. no., 02-024-1A) and pre-blocked with DMEM containing 0.5% BSA (albumin bovine fraction-V; NeoFroxx, Einhausen, Germany; cat. no., GRM105-5G) without supplements at 37˚C and 5% CO2 for 1 h. The Ph.D.C7C™ Phage Display Library (1e11 PFU) was then introduced to the cells. Non-binding phages were then discarded, and the cells were washed 5 times in cold PBS. The phages binding to the cells were eluted with 1 ml Glycine/HCl (pH 2.2) with 1 mg/ml BSA for 20 min at room temperature while being gently shaken. The eluted phages were collected and neutralized with 0.15 ml 1 M Tris-HCl (pH 9.1). A total of 3 rounds of panning were performed for each sample. A peptide sequence (CDNVAQSVC), termed D7, was identified through phage display technology using C57BL/6 mouse BMMSCs. Subsequent analysis suggested that the identified loop-constrained heptapeptide exhibited a high specific affinity for mouse BMMSCs. Due to this specific affinity for BMMSCs, the present study provides a selective method to improve mesenchymal stem cell (MSC) based tissue engineering (TE) strategies for the treatment of osteonecrosis of the femoral head (ONFH). 3376 QVNGLGERSQQM(9) GLRTMQSPNSFY(1) VWDSGLRQSRPA(1) 3 Phage display (common panning) 4 PMID:30519000 Complement decay-accelerating factor Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Adherent HeLa cells were washed with serum-free RPMI-1640 and blocked with 16% culture medium containing 0.1% BSA for 1 hour, and then added to the stock solution of Ph.D.-12 phage peptide library (titer: 1e11 pfu/mL) for 1 hour. After washing on ice with a pre-cooled 0.1% PBST at 4°C to remove non-cell-bound phage, the phage bound to the cell surface was eluted on ice with a glycine buffer (pH 2.2) pre-cooled at 4°C immediately, and then added to a centrifuge tube pre-filled with 250 μL Tris buffer (pH 2.2). The effects of the peptide (QVNGLGERSQQM) on the proliferation and apoptosis of HeLa and SiHa cells were determined by Cell Counting Kit-8 (CCK-8), flow cytometry, and TUNEL assay, respectively, which showed that the peptide can bind to the CD55 molecule on the surface of cervical cancer HeLa and SiHa cells as a ligand peptide. It can also effectively inhibit the proliferation of cervical cancer cells and induce cell apoptosis. 3377 WLNMVWGVL(6) GRWAVLVNL(3) GVAVIFDFV(2) DTTVVVRMM(2) TPGGIVYRL(1) WSDSIVVHH(1) FQRFIVVAA(1) PGGLILLAD(1) SGWLILRDL(1) GDGCILVWR(1) TPGGILTWR(1) TPGGILFFR(1) ELGFIFLEV(1) GVPEIWMAF(1) GSVSIWYAD(1) GGWVIWLGL(1) TPGGIWLVE(1) PGGGIMSLV(1) LWIRIQMRH(1) TPGGITELY(1) TYRDITWHH(1) LKLLISDEH(1) FLNWISHHH(1) WFWFINEEH(1) LFTKIEHHH(1) RAVYIDDWL(1) ARRLIRYMT(1) GGERALFAS(1) FYYWAATRH(1) WFPYAWGVQ(1) TMELARFML(1) PGGQATYLE(1) WTGWYSCF(1) TWMPYFGG(1) GPDMGDTW(1) LDGELWFL(1) TPLLVWSHH(1) TPGGVWLRD(1) GGVGVWWSD(1) CPYSVWRPH(1) GVVEVWRAG(1) TPGGVYYRL(1) GVPGVYFVE(1) GKMVVYLQV(1) PYMDVYAHH(1) GFFRVYVEL(1) GGGGVFAMF(1) PLVDVFSSW(1) GVSTVFLSM(1) GGVLVFWSK(1) GGGAVGYSA(1) MDPGVGYWG(1) TPGGVGFWW(1) PGGLVAFNF(1) KYLAVAHHH(1) GVWCVAVLP(1) PGGRVLSFA(1) PGGRVLLVE(1) GGPTVLFGV(1) WAETVLFAH(1) SVSVVLWLH(1) LDYAVLVLY(1) TPGGVLELA(1) PGGEVLRLL(1) GWHLVPLVQ(1) EQYLVVESH(1) GSEVVMLLD(1) SPLWVMGHH(1) GGRVVMFAS(1) GYLTVNFST(1) EVDFVSFHH(1) GGAWVSYFG(1) GGTTVSQFL(1) FAAEVSHHH(1) PGGEVSLVL(1) GLSVVSAFM(1) GGLAVSAFL(1) VLLAVTLSE(1) PGGAVTFVF(1) YWGYVTHHH(1) PGGFVKAGM(1) CALAVKYHH(1) NGPAVHHHH(1) 83 Phage display (common panning) 8 PMID:30521603 Mast cell protease 2 Not determined. Not determined. Not determined. X9 T7 phage display library An aliquot of the amplified phages (~e9 pfu) were bound to 100 μl Ni-NTA beads by their His6-tags for 1 hr at 4°C under gentle agitation. Unbound phages were removed by washing ten times in 1.5 ml 1 M NaCl, 0.1% Tween-20 in PBS, pH 7.2, and two subsequent washes with 1.5 ml PBS. The beads were finally resuspended in 1 ml PBS. Activated mast cell protease 2 (~0.1 μg) was added to the resuspended beads and left to digest susceptible phage nonapeptides under gentle agitation at room temperature for 2 hours. PBS without protease was used as control. Phages with a random peptide that was susceptible to protease cleavage were released from the Ni-NTA matrix, and the supernatant containing these phages was recovered. To ensure that all of the released phages were recovered the beads were resuspended in 100 μl PBS (pH 7.2) and the supernatant, after mixing and centrifugation, was added to the first supernatant. To ensure that the His6-tags had been hydrolyzed on all phages recovered after protease digestion, 15 μl fresh Ni-NTA agarose beads were added to the combined phage supernatant and the mixture agitated for 15 min followed by centrifugation. 3378 GWSLFLEVL(3) GMATFLELH(3) AVLLFAGIL(3) WEVMFLDFH(2) MSLFWSDWH(2) WSVLFNWGH(2) TPGGYILML(2) GWGVFLDVG(1) GGVLFLDYF(1) GQALFLDIF(1) LIMLFLEEE(1) GIVGFLEVL(1) GWEFFLDAE(1) GVVSFLEMC(1) GWVSFLEPR(1) RATSFLEWH(1) VVMSFLDFS(1) GPVLWLEVM(1) AWVPWLDLV(1) GWVSYLEGG(1) GWMLYLEWR(1) LFVNFGEHH(1) WYVYFGELD(1) GATTFTDMV(1) GLETFIEVV(1) TPGGFVELY(1) WMTSFSDVM(1) WVVSFSETP(1) GIVVWSDVL(1) SVELWSEVL(1) AWVPWSEVH(1) VERFWSVEH(1) PVLLFMDFP(1) GGVLFMDEI(1) GWSAFMEFT(1) DTRFFQEWL(1) GFLSFWDLA(1) TWVSFFDLH(1) GVISFFEFV(1) GWVTWWDIG(1) EWMTYWEEG(1) EVIKWWDFH(1) GVVQYWGGD(1) WGVFYWEHH(1) GYALWWAIE(1) GWTVWFELV(1) SSTLYLIFE(1) YFWLWDHHH(1) WFLLWEPHH(1) GLWGWEGFR(1) TPGGFRGEG(1) RVYLWIGEH(1) EMELFWTHH(1) ECVVWGRHH(1) VAALWALFH(1) QGGKWALAY(1) ATYRWAASG(1) GPVVFLVRY(1) VLGIFLAVR(1) GSGQFLLFL(1) WAVWFLLIP(1) GYVTFLLIM(1) GWSTYLSYS(1) GAGFSTGW(1) VLYWFSSSW(1) LGITFMILH(1) LLWGFMFHH(1) VLVWWAKRA(1) GGNWWGVAT(1) GGLWWSYRS(1) TPGGWCQVQ(1) PGGDFRPAV(1) TPGGFRIVT(1) GGWVFRSLG(1) LFIFWHAVH(1) 75 Phage display (common panning) 5 PMID:30521603 Mast cell protease 1 Not determined. Not determined. Not determined. pComb M13 phage display library An aliquot of the amplified phages (~e9 pfu) were bound to 100 μl Ni-NTA beads by their His6-tags for 1 hr at 4°C under gentle agitation. Unbound phages were removed by washing ten times in 1.5 ml 1 M NaCl, 0.1% Tween-20 in PBS, pH 7.2, and two subsequent washes with 1.5 ml PBS. The beads were finally resuspended in 1 ml PBS. Activated mast cell protease 1 (~0.1 μg) was added to the resuspended beads and left to digest susceptible phage nonapeptides under gentle agitation at room temperature for 2 hours. PBS without protease was used as control. Phages with a random peptide that was susceptible to protease cleavage were released from the Ni-NTA matrix, and the supernatant containing these phages was recovered. To ensure that all of the released phages were recovered the beads were resuspended in 100 μl PBS (pH 7.2) and the supernatant, after mixing and centrifugation, was added to the first supernatant. To ensure that the His6-tags had been hydrolyzed on all phages recovered after protease digestion, 15 μl fresh Ni-NTA agarose beads were added to the combined phage supernatant and the mixture agitated for 15 min followed by centrifugation. 3379 LSFMPPE(2)[0.27] FSFIAPE(2)[n.a.] FSFLPPE(1)[0.57] FSFLAPE(1)[n.a.] FSFMVPE(1)[n.a.] LSFVPPE(1)[n.a.] FPWVPEL(1)[n.a.] FPWIPEE(1)[n.a.] [FL]SF[PAV]PE 8 Phage display (common panning) 3 PMID:30394584 Anti-GSFLSPEHQRVQ polyclonal antibody GSFLSPEHQRVQ Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The fraction of N-terminal specific antibodies obtained after purification was evaluated usingenzyme linked immunosorbent assay (ELISA). Biotinylated human ghrelin (Bachem, Bubendorf, Switzerland) was immobilized to streptavidin coated microtiter plates (StreptaWell, Roche, Basel, Switzerland) and affinity purified anti N-terminal antibodiesobtained from 5 µg of purified antibodies or unpurified (5 µg) were added. Binding was detected by secondary goat anti rabbit horseradish peroxidase (HRP)-conjugated antibodies (Merck Millipore, Darmstadt, Germany) diluted 1:5000 in 0.1% bovine serum albumin (BSA) and reagent 3,3',5,5'-tetramethylbenzidine (TMB) (Thermo Fisher Scientific, Waltham, MA, USA). The absorbance at 450 nm (A (405 nm)) was read using the Tecan Safire (Tecan, Grödig, Austria) microtiter plate reader. A (405 nm) values were preproduced from Figure 1B and shown. Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). Peptides were selected and tested using calcium screening assays. The most effective competitive antagonist FSFLPPE was further tested in vivo. Administration of the peptide produced no significant effect on either food intake or growth hormone (GH) release. Surprisingly, when co-administered with ghrelin the peptide significantly enhanced GH secretion and c-Fos expression. The evidence indicates that FSFLPPE might act as an ago-allosteric modulator. 3380 TVDDMAAMDGGH(1)[0.20] GLIDGMMFYQRG(1)[0.51] NIFASDTTKSTW(1)[0.31] WSQAYIAQFAGR(1)[n.a.] LPHRNLFEIEGR(1)[n.a.] 5 Phage display (common panning) 3 PMID:30394584 Anti N-terminal appetite-regulating hormone[24-51] polyclonal antibody N-terminal appetite-regulating hormone[24-51] Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The fraction of N-terminal specific antibodies obtained after purification was evaluated usingenzyme linked immunosorbent assay (ELISA). Biotinylated human ghrelin (Bachem, Bubendorf, Switzerland) was immobilized to streptavidin coated microtiter plates (StreptaWell, Roche, Basel, Switzerland) and affinity purified anti N-terminal antibodiesobtained from 5 µg of purified antibodies or unpurified (5 µg) were added. Binding was detected by secondary goat anti rabbit horseradish peroxidase (HRP)-conjugated antibodies (Merck Millipore, Darmstadt, Germany) diluted 1:5000 in 0.1% bovine serum albumin (BSA) and reagent 3,3',5,5'-tetramethylbenzidine (TMB) (Thermo Fisher Scientific, Waltham, MA, USA). The absorbance at 450 nm (A (405 nm)) was read using the Tecan Safire (Tecan, Grödig, Austria) microtiter plate reader. A (405 nm) values were preproduced from Figure 1B and shown. Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3381 NIFASAD(1)[0.31] GLVDVLF(1)[0.52] MNYREIY(1)[0.43] GLVDALY(1)[n.a.] MNRHIPA(1)[n.a.] GLFALERSK(1)[n.a.] 6 Phage display (common panning) 3 PMID:30394584 Anti N-terminal appetite-regulating hormone[24-51] polyclonal antibody N-terminal appetite-regulating hormone[24-51] Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The fraction of N-terminal specific antibodies obtained after purification was evaluated usingenzyme linked immunosorbent assay (ELISA). Biotinylated human ghrelin (Bachem, Bubendorf, Switzerland) was immobilized to streptavidin coated microtiter plates (StreptaWell, Roche, Basel, Switzerland) and affinity purified anti N-terminal antibodiesobtained from 5 µg of purified antibodies or unpurified (5 µg) were added. Binding was detected by secondary goat anti rabbit horseradish peroxidase (HRP)-conjugated antibodies (Merck Millipore, Darmstadt, Germany) diluted 1:5000 in 0.1% bovine serum albumin (BSA) and reagent 3,3',5,5'-tetramethylbenzidine (TMB) (Thermo Fisher Scientific, Waltham, MA, USA). The absorbance at 450 nm (A (405 nm)) was read using the Tecan Safire (Tecan, Grödig, Austria) microtiter plate reader. A (405 nm) values were preproduced from Figure 1B and shown. Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3382 CIALNNNEC(1)[0.15] CSPKHQGSC(1)[0.15] CSPKHWGPC(1)[0.30] CIILNNNNC(1)[n.a.] 4 Phage display (common panning) 3 PMID:30394584 Anti N-terminal appetite-regulating hormone[24-51] polyclonal antibody N-terminal appetite-regulating hormone[24-51] Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The fraction of N-terminal specific antibodies obtained after purification was evaluated usingenzyme linked immunosorbent assay (ELISA). Biotinylated human ghrelin (Bachem, Bubendorf, Switzerland) was immobilized to streptavidin coated microtiter plates (StreptaWell, Roche, Basel, Switzerland) and affinity purified anti N-terminal antibodiesobtained from 5 µg of purified antibodies or unpurified (5 µg) were added. Binding was detected by secondary goat anti rabbit horseradish peroxidase (HRP)-conjugated antibodies (Merck Millipore, Darmstadt, Germany) diluted 1:5000 in 0.1% bovine serum albumin (BSA) and reagent 3,3',5,5'-tetramethylbenzidine (TMB) (Thermo Fisher Scientific, Waltham, MA, USA). The absorbance at 450 nm (A (405 nm)) was read using the Tecan Safire (Tecan, Grödig, Austria) microtiter plate reader. A (405 nm) values were preproduced from Figure 1B and shown. Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3383 DERAYWFKSDLR(2) QIPERAYWVWSP(1) HALEDSKPPRTK(1) SAHNLKEQLKPP(1) 4 Phage display (common panning) 3 PMID:30394584 Anti C-terminal appetite-regulating hormone[24-51] polyclonal antibody C-terminal appetite-regulating hormone[24-51]Growth hormone secretagogue Growth hormone-releasing peptide Motilin-related peptide Protein M46 Not determined. Not determined. Ph.D.-12 phage display library (X12) Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3384 ASVQERK(2) AEKIQPR(2) TQKIQPR(1) 3 Phage display (common panning) 3 PMID:30394584 Anti C-terminal appetite-regulating hormone[24-51] polyclonal antibody C-terminal appetite-regulating hormone[24-51]Growth hormone secretagogue Growth hormone-releasing peptide Motilin-related peptide Protein M46 Not determined. Not determined. Ph.D.-7 phage display library (X7) Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3385 CKSDIQRSC(1) CIQKGHPMC(1) CIKFQPAQC(1) CVRYQPHQC(1) CVKLGKLYC(1) 5 Phage display (common panning) 3 PMID:30394584 Anti C-terminal appetite-regulating hormone[24-51] polyclonal antibody C-terminal appetite-regulating hormone[24-51]Growth hormone secretagogue Growth hormone-releasing peptide Motilin-related peptide Protein M46 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3386 SGVYKVAYDWQH(11) GLHTSATNLYLH(2) TVLSHPSTATLI(1) QQRPYVQDLRLI(1) 4 Phage display (subtractive panning) 4 PMID:30387183 BDNF/NT-3 growth factors receptor (EC:2.7.10.1)[1-430] Brain-derived neurotrophic factor, BDNF Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The phage particles were added to microtiter wells coated with 10 µg/mL of NTRK2, p75NTR (as nonspecific bindings), or bovineserum albumin (BSA; as a negative control) in coating buffer. The plate was washed three times after 1 hour incubation and 50µL of an anti-M13 antibody-horseradish peroxidase conjugate (GE Healthcare) was added. Finally, 50 µL of a substrate chromogen (TMB) was added and the plate was incubated at room temperature in the dark. Absorbance at 450nm was measured using Multiskan ELISA reader. The specificities and affinities of phage-displayed peptides were shown in Figure 3. In the first round, the Ph.D‐12 peptide library (New England Biolabs, Herts, UK) was added to 600 ng of recombinant BDNF/NT-3 growth factors receptor in 200 µL of tris‐buffered saline Tween 20 (TBS/T; 0.1% [vol/vol] Tween 20) and incubated for 2 hours. Then, 50 μL of protein G magnetic beads (Protein G Mag Sepharose; GE Healthcare UK Ltd, Little Chalfont, Buckinghamshire, UK) were added. After incubation and magnetic separation, the supernatant was removed (containing unbound phage). The magnetic beads were washed eight times with 1mL of TBS/T (0.1%) to remove weakly‐or nonspecifically‐bound phage. In the second round, one part of the amplified phage stock from the first round was preincubated with the beads in the absence of target to exclude protein G‐binding phage particles. Then, the unbound phage fraction was incubated with recombinant human BDNF/NT-3 growth factors receptor and specifically. The stringency of panning was increased in the third round of panning using reduced concentration of target protein and increased concentration of detergent. Also, a further round of biopanning was carried out and the phage particles were preincubated with recombinant human p75NTR ((nerve growth factor receptor/TNFRSF16 Fc Chimera; R&D System, Minneapolis, MN). The conserved residues of loop2 of brain-derived neurotrophic factor (BNDF) including S32‐GTVLKVPVQ‐L49 shared the most identity with the peptide 1 sequence (SGVYKVAYDWQH). Also, GLAM2SCAN algorithm, RRID:SCR_00178334 introduced peptide 1 as a motif due to the most frequency and similarity with BDNF. In silico molecular docking showed strong interactions between the peptide three dimensional models and the surface residues of BDNF/NT-3 growth factors receptor at the IgC2 domain. A consensus peptide sequence was then synthesized to generate a mimetic construct (named as RNYK). The affinity binding and function of this construct was confirmed by testing against the native structure of BDNF/NT-3 growth factors receptor in SH-SY5Y cells in vitro using flow-cytometry and MTT assays, respectively. RNYK at 5 ng/mL prevented neuronal degeneration of all trans-retinoic acid-treated SH-SY5Y with equal efficacy to or even better than BDNF at 50 ng/mL. 3387 NWTTLSRSVNWP WIYDTTRVIVPG HAMSPVFLSKYA LVPSILGATFIH DISASLQSNRWH AAGTFLMSMMSR NNLPTSRTLAGN GNNPLHVHHDKR HHLRIPYALDQT GTGAALAKVSEA IKPVRALYTLAD GTIRTSFWHTNT GVHSVFAPLTPN 13 Phage display (common panning) 3 PMID:30132552 Endoglin Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured using a VICTOR X5 Multilabel plate reader (PerkinElmer, Inc., Singapore) at 490 nm, and the results are reported as the optical density. The binding activity of 16 monoclonal M13 phages to human recombinant CD105 protein and negative control blocking buffer was shown in Table II. The recombinant human CD105 protein (R&D Systems, Inc., Minneapolis, MN) at 100 µg/ml in sterile phosphate-buffered saline (PBS) was added to individual sterilised MaxiSorp plates (Thermo Fisher Scientific, Inc., Waltham, MA, USA) and incubated overnight at 4˚C. The plate was washed six times with Tris-buffered saline/0.1% Tween-20 (TBST) and then blocked for 1 h at 4˚C using 1% bovine serum albumin (BSA) in PBS. M13 phage display libraries were allowed to bind for 1 h at room temperature, and the unbound phages were washed away 10 times with TBST. Subsequently, the binding phages were eluted by an elution buffer, and the eluted phages were then amplified for another round of biopanning. A novel non-antibody-binding protein (WIYDTTRVIVPG) with a high affinity for recombinant human CD105 was identified by M13 phage biopanning. Fluorescence microscopy, flow cytometry and in vivo animal imaging were used to confirm the visualization effect of the novel peptide to the CD105-positive MNNG/HOS cell line in vivo and in vitro. Immunofluorescence of tissue sections was also used to identify the target efficiency of the peptide in tumour sections derived from an MNNG/HOS xenograft tumour model and osteosarcoma patients. This peptide specifically was found to bind to the CD105-positive osteosarcoma MNNG/HOS cell line and the osteosarcoma cells in the histological sections derived from an MNNG/HOS xenograft model and osteosarcoma patients in vitro. This peptide also successfully labelled an animal osteosarcoma xenograft model in vivo. Taken together, a novel peptide that can be used as a potent agent for CD105 molecular targeting was identified, and this peptide can be applied for osteosarcoma visualization in vitro and in vivo. 3388 ACLHFGVFCSMQHDNCG(3)[+++] ACLHFRDRCGSLQSGCG(2)[+] ACLHWTDSCLHYTQLCG(2)[+] ACMNYGVLCQFVTSTCG(2)[n.a.] ACLHYSHLCQTKMDACG(1)[n.a.] ACLHFAHSCEQQAMRCG(1)[n.a.] ACLHYAQSCDGQHSVCG(1)[n.a.] ACLHFQGMCSELMEGCG(1)[+] ACLHFGAHCSQVVTDCG(1)[n.a.] ACLHGRGECSDMDFQCG(1)[++] ACLHGRQGCSATTDNCG(1)[n.a.] ACLHGPGLCQGLSYPCG(1)[n.a.] ACLSFGTNCDSQVFTCG(1)[n.a.] ACFSYGQLCDRAVYKCG(1)[n.a.] ACLNYGQWCDRMSLGCG(1)[++] ACYHFGLHCSQTKILCG(1)[n.a.] ACFHYGTGCTREQLFCG(1)[n.a.] ACWHGPPGCDSTFYMCG(1)[n.a.] 18 Phage display (common panning) 3 PMID:30411004 Rabbit anti-goat IgG (H + L) superclonal secondary antibody Not determined. Not determined. Not determined. CX6CX6C phage display library Six individual selected sequences representing different sequence motifs were synthesized and functionalized with BiotinSH. As a negative control, a sequence (JK38) sharing no observable similarity to any of the chosen motifs was used. The BiotinSH-functionalized peptides were captured by streptavidin immobilized on magnetic beads. After incubation with the target protein and five washing cycles, each pulldown experiment was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The phage-encoded peptide library was selectively modified by the two-step modification under optimized conditions using NATBA as a linker and FlcSH as an additive, which was then subjected to three iterative rounds of affinity selection against the model target (rabbit anti-goat IgG (H + L) Superclonal Secondary Antibody). Briefly, biotinylated Rabbit anti-Goat IgG (H + L) Superclonal Secondary Antibody (ThermoFischer) was immobilized on magnetic streptavidin beads (Dynabeads M-280 Streptavidin, Invitrogen). The bead suspension and phage suspension were then mixed together and incubated for 30 min on a rotating wheel at room temperature. Second and third rounds of panning were performed following the same procedure but using in the second round instead of streptavidin beads neutravidin-coated magnetic beads. We were pleased to find strong consensus motifs within the resulting peptides sequences as aclear indication for target-specific binding. Most of the similarities are found in the N-terminal part of the first loop, consisting predominately of Leu, His, and Phe/Tyr. In the second loop, the highest conserved position is likewise the most N-terminal amino acid next to the central Cys, showing a strong preference for polaror acidic residues (Ser, Asp, Gln). None of the isolated sequences were found with an abundance higher than 3. 3389 YKLKIRTPQ YKLKVRTPQ YKLRIRTPQ YKLEIRTPQ YRLKIRTPQ YKLKIRTPR YKLKIQTPQ YKLKTRTPQ YELKIRTPQ YKLKIRAPQ YKLKIRIPQ HKLKIRTPQ NKLKIRTPQ YKPKIRTPQ YKLKIRTPL CKLKIRTPQ YKLKIRTLQ YKLKIRTSQ LELLKASRW LELLRASRW LELLEASRW LELLKVSRW LELLKAPRW LELLKALRW LESLKASRW LELPKASRW LGLLKASRW PELLKASRW LELLKASRR LQLLALSRT LRLLALSRT LQLLALSRA LQLLAPSRT PQLLALSRT LQLPALSRT LELVRRSPV LELARRSPV LELVRRSPA LELVHRSPV LELVRHSPV LELVRCSPV LEPVRRSPV LELVRRSSV LELVCRSPV LALIKRQPL LALLMRQRP LAMLTRGRP YKLKIRTPQ, LELLX(2)SRX 47 Phage display (common panning) 3 PMID:30373771 Ubiquitin carboxyl-terminal hydrolase 11 [1–244 or 24–244] Not determined. Not determined. Not determined. X9 PC89-based phage display library Proteins were immobilized on cobalt beads (Dynabeads) (solution phase panning). A total of 100 μg of USP11_DU was immobilized for each round. Bound phage were washed 10–20 times with PBS (10 mM phosphate buffer, pH 7.2–7.4, with 150 mM NaCl) plus 0.1% (v/v) Tween (PBST) and then PBS. Bound phage were eluted from the beads with 100 mM triethylamine and neutralized in 1 M Tris, pH 7.4. We discovered unique USP11-interacting peptide motifs. Isothermal titration calorimetry disclosed that the highest affinity peptides (AEGEFYKLKIRTPQ, KD of ~10 μM) exhibit exclusive selectivity for USP11 over USP4 and USP15 in vitro. Furthermore, a crystal structure of a USP11–peptide complex revealed a previously unknown binding site in USP11’s noncatalytic ubiquitin-like (UBL) region. This site interacted with a helical motif and is absent in USP4 and USP15. Reporter assays using USP11-WT versus a binding pocket–deficient double mutant disclosed that this binding site modulates USP11’s function in homologous recombination–mediated DNA repair. The highest affinity USP11 peptide binder fused to a cellular delivery sequence induced significant nuclear localization and cell cycle arrest in S phase, affecting the viability of different mammalian cell lines. The USP11 peptide ligands and the paralog-specific functional site in USP11 identified here provide a framework for the development of new biochemical tools and therapeutic agents. 3390 SDEEFNFINSAFDVADVNWYKNN[3.2] 1 Phage display (subtractive panning) 3 PMID:30203962 The variant of photoactive yellow protein (PYP) in the light-state Not determined. Not determined. Not determined. XDXXFNXINXAXXVXXVNXXKNX phage display library NMR spectroscopy All NMR experiments were carried out at CSICOMP (Department of Chemistry, University of Toronto) on an Agilent DD2 700 MHz spectrometer equipped with an HFCN cold probe. The Kd (μM) value was shown. Proteins were immobilized on neutravidin-coated plates. For selecting ligtht-state of cPYP binders, the phage libraries were depleted by allowing binding to dark-adapted protein (cPYP-Dark) for 1 h prior to transferring the unbound phage to the light-adapted plate for positive selection. The phage bound to the light-adapted protein (cPYP-Light) was washed 8 times with PT buffer, eluted and amplified overnight and the panning process was repeated 3 times. For panning rounds 2 and 3, a lower concentration of phage was used as the high-avidity of display often results in very high elute concentration. 3391 QDYLFNRINEAHFVNQVNVRKNV[20] 1 Phage display (subtractive panning) 3 PMID:30203962 The variant of AsLOV2 in the dark-state Not determined. Not determined. Not determined. XDXXFNXINXAXXVXXVNXXKNX phage display library NMR spectroscopy All NMR experiments were carried out at CSICOMP (Department of Chemistry, University of Toronto) on an Agilent DD2 700 MHz spectrometer equipped with an HFCN cold probe. The Kd (μM) value was shown. Proteins were immobilized on neutravidin-coated plates. For selecting dark-state of AsLOV2 binders, the phage libraries were depleted by allowing binding to light-adapted protein (AsLOV2-Light) for 1 h prior to transferring the unbound phage to the dark-adapted plate for positive selection. The phage bound to the dark-adapted protein (AsLOV2-Dark) was washed 8 times with PT buffer, eluted and amplified overnight and the panning process was repeated 3 times. For panning rounds 2 and 3, a lower concentration of phage was used as the high-avidity of display often results in very high elute concentration. 3392 QVDTQGENAVKV(189) PTVYHPELYQKA(187) AVMRQQTDELRL(186) AVHSNLFPGQPD(185) DPSDVLTLPFPR(183) FQFASGNEANET(181) WEIANPYWDGSE(170) VTVRENSPRKLA(166) YPNLLLLASVDV(166) QGVSDIHSRNLT(159) APAQPAESIHAY(155) RVTAPRPEFSTL(147) LPRVPPPVHSTT(143) ALSKTFEVAPLH(142) AYPSYLTSDGYH(141) IDTQYPSAMTLT(140) DIHRHVVGARTL(136) TTMRIAFHQLHT(134) RGELTNSGKARE(134) HGRFPLTSDVPT(123) SMPSMLFDTGED(121) ACAATPLNCGG(119) QIRDRIHDNELE(116) VETIPPLRYSDP(110) SENKNCNAGSLT(102) QPPHIHSALTLM(101) VAGTLPAPSPSY(90) NLGNYNDKEAVN(84) HDWSSKTETNAT(84) FMNTHDRADLSI(81) LLKHIEVSLPLA(80) QWYHRSDGGGSA(70) AINSTTGKRNVV(61) LACAVTGLICGG(59) RKAHQEKDSPRI(51) 35 Phage display (common panning) 1 PMID:30249752 Serum from patients with wasp sting injuries Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A total of 4356 peptides were achieved in the 3-h group, and 4408 peptides in 4-day group. We compared the peptides between the two groups, and 35 specific peptides were achieved in the 4-day group. Amongst them, twelve peptide epitopes were matched with nine wasp venom proteins, namely, vitellogenin precursor, hexamerin 70b precursor, venom carboxylesterase-6 precursor, MRJP5, major royal jelly protein 8 precursor, venom acid phosphatase Acph-1 precursor, phospholipase A2, venom serine protease 34 precursor, and major royal jelly protein 9 precursor. The changes in serum IgM antibodies induced by wasp venom were confirmed by ELISA based on the 12 peptide epitopes. 3393 ADGVGDAESRTR[5.27e-8] 1 Phage display (common panning) 3 PMID:30260652 Chitosan Not determined. Not determined. Not determined. SUT12 M13 phage display library Surface plasmon resonance (SPR) Interactions between peptide and chitosan were quantified by SPR (Nicoya Lifescience, Waterloo, Canada). The binding constant (KD, M) was determined and shown. For biopanning of chitosan binding peptide, TBST (50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 0.1% Tween 20) was used to wash each well three times before adding 10 μl of the 12-amino acid-long random peptide library (SUT12, ~10^10 pfu) in 90 μl of TBST. The incubation time of the phage stock in the wells decreased with increasing rounds of selection. Following incubation at 25℃ for 4 h, the wells were washed for at least five times with TBST to remove unbound phages. In the first round, each well was washed for 6 times with no incubation. In the following rounds, the wells were washed twelve times with incubation for amounts of time. The bound phages were eluted by acid elution (50 μl of 50 mM glycine–HCl, pH 2.0). Then phosphate buffer (pH 7.5) was applied for neutralization of solutions. plates. Three rounds of biopanning were undertaken with chitosan according to the above mentioned methods. The chitosan-binding peptide (ADGVGDAESRTR) was used as the targeting moiety, followed by conjugation to the surface of poly(lactic-co-glycolic acid) nanoparticles as the drug carrier, which was then incubated with free chitosan. The noncovalently bound chitosan adheres to mucus layers and significantly enhances penetration of nanoparticles through the oral absorption barrier into circulation and then re-exposed the targeting ligand for later recognition of the fungal pathogen at the site of infection. 3394 FNKWMDCLSVTH 1 Phage display (common panning) 3 PMID:30417023 Anti-histamine H4 receptor (H4R) monoclonal antibody Histamine H4 receptor, H4R Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The bingding affinity of the identified phages to human H4R mAb was examined by ELISA. The absorbance at 450 nm (OD450) was measured, which was reproduced from Figure 1 and shown. 96-well microtiter plates were coated with affinity-purified anti-H4R antibody (40 μg/well) in sodium bicarbonate buffer (pH 8.6) and incubated overnight at 4°C. Blocking buffer consisting of 1% (w/v) BSA/Tris-buffered saline (TBS) was added to each well and incubated for 2 hr at room temperature, followed by 1 hr at 37°C using the phage library (total of 2e11 phages). The wells were washed six times with TBS and Tween 20 (TBST). Unbound phages were discarded and the wells were washed five times with TBST. Bound phages were eluted by adding elution buffer (0.2 M glycine-HCl in 1 mg/mL BSA [pH 2.2]) to each well, and the plates were gently rocked for 12 min at room temperature. The eluate was neutralized with 1 M Tris-HCl (pH 9.1). The eluate was amplified and precipitated with 20% (w/v) PEG-8000. A second round of panning was conducted using the first round of amplified eluate as the input phages. The entire screening protocol was repeated for a total of three rounds of panning. The peptide FNKWMDCLSVTH, designated as P-FN12, was bound by H4R monoclonal antibody (mAb) with high affinity. Moreover, the P-FN12 + CTB@Lipo-formulated vaccine, used as nasal drops, decreased allergic symptoms such as sneezing and nasal rubbing in a rat model. 3395 KRLTPLSPDHYD(14/30)[4.8 ± 1.0] QLKLHNKGLISS(1/30)[3.2 ± 1.5] GWSSREASQEGN(1/30)[NT] NGVFAMLFPMQS(1/30)[NT] YGILARDNDSLG(1/30)[NT] VAYEGSYFAVYQ(1/30)[NT] SIDSGYRPGVRY(1/30)[NT] ADHWWSYAYQKP(1/30)[NT] VVVSSLSAPYKD(1/30)[NT] VPAPPISSASD(1/30)[NT] ITTPDWELMRGM(1/30)[NT] GIVEPSDRSTRG(1/30)[NT] AFDSPDNPVASN(1/30)[NT] EYHDYESANAAD(1/30)[NT] 14 Phage display (subtractive panning) 3 PMID:30344835 Ovarian cancer tissue slice Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Flow Cytometry Peptides were synthesized (>98% purity) and conjugated with FAM (excitation wavelength = 490 nm, emission wavelength = 525 nm) at the 5'- and carboxyl termini (Protech Co., Ltd., Taiwan). These fluorescently-labeled peptides were then used for fluorescence microscopy analysis and flow cytometric assays (BD Accuri™ C6, Becton, Dickinson and Company, USA). The resulting fluorescence intensity of the affinity reagents was further used to calculate the equilibrium dissociation constant (Kd, nM) with Prism software (GraphPad Software, USA). Selection was performed against ovarian cancer tissue slides by the integrated microfluidic system capable of automating SELEX and phage display technology. Briefly, an M13 phage library was incubated with normal tissues (“negative selection” step), and then the unbound M13 phages were incubated with cancer tissues (“positive selection” step). There were three rounds of both positive and negative panning involved for the entire screening process. 3396 GTGSQAS(7)[2.83] WTTSVGT(1)[NT] TVNFKLY(1)[NT] GVEGHKP(1)[NT] FTAERYY(1)[NT] QQTNWSL(1)[NT] 6 Phage display (common panning) 5 PMID:30592604 Gold Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Surface plasmon resonance (SPR) To evaluate the binding affinity of the peptide GTGSQAS against the gold surface, the SPR analysis was performed. The binding affinities (Kd, μM) of all peptides were determined. The Kd values of previously reported gold-binding peptides VSGSSPDS and KHKHWHWG are 2.28 μM and 3.67 μM, respectively. The biopanning process was performed using the gold-patterned microfluidic chip with NEB Ph.D.-7 phage display peptide library kit. Ten microliters of phage solution (2e12 pfu mL−1) in 0.1 vol % PBST (PBS (pH 7.4) containing 0.1 vol % Tween 20) was loaded into the microfluidic biopanning platform at a flow rate of 1 μL min−1 at room temperature. Then PBST (0.1 vol % Tween 20 in the first round and 0.5 vol % Tween 20 from the second round) was introduced at 0.5 mL min−1 for 30 min to remove nonspecifically or weakly bound phages. The remaining high-affinity phages were eluted by injecting an elution buffer (0.2 M glycine-HCl (pH 2.2) with 1 mg mL−1 BSA) at 0.1 mL min−1 for 10 min, and the eluate was neutralized with 150 μL of 1 M Tris-HCl (pH 9.1). Five rounds of biopanning were performed, and the eluted phages from each round were amplified by incubation with ER2738 host cells. Finally, the phage peptide sequence was analyzed by sequencing the phage plasmid DNA. The surface plasmon resonance analysis shows that the binding affinity of the identified GTGSQAS is comparable to previously reported gold-binding peptides (GBPs). Moreover, molecular dynamics simulations are performed to understand its binding affinity against the gold surface in detail. Theoretical calculations suggest that the association and dissociation rates of the GBPs depend on their sequence-dependent conformations and interactions with the gold surface. 3397 GTGSQAS(4) GVEGHKP(3) QLAVAPS(2) MNSNIPI(2) TVNFKLY(2) ANPKNFS(1) FSGGGNH(1) ELWSLYD(1) NLQPPAY(1) RHFIKEL(1) NDLMNRA(1) 11 Phage display (common panning) 4 PMID:30592604 Gold Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The biopanning process was performed using the gold-patterned microfluidic chip with NEB Ph.D.-7 phage display peptide library kit. Ten microliters of phage solution (2e12 pfu mL−1) in 0.1 vol % PBST (PBS (pH 7.4) containing 0.1 vol % Tween 20) was loaded into the microfluidic biopanning platform at a flow rate of 1 μL min−1 at room temperature. Then PBST (0.1 vol % Tween 20 in the first round and 0.5 vol % Tween 20 from the second round) was introduced at 0.5 mL min−1 for 30 min to remove nonspecifically or weakly bound phages. The remaining high-affinity phages were eluted by injecting an elution buffer (0.2 M glycine-HCl (pH 2.2) with 1 mg mL−1 BSA) at 0.1 mL min−1 for 10 min, and the eluate was neutralized with 150 μL of 1 M Tris-HCl (pH 9.1). As the rounds progress, the proportion of high-affinity bacteriophages increases and fewer bacteriophages are removed from the surface after the washing step. Thus, the number of eluted bacteriophages increases as the rounds continue. 3398 HAAWHFI(2) VSRDTPQ(2) GTGSQAS(1) GASESYL(1) HSHTLTW(1) NHFTLNQ(1) YNGSANQ(1) SSQSLRE(1) TVNFKLY(1) QQTNWSL(1) YSEPAVT(1) SYTDLLR(1) GQSEKHL(1) TIVGAKD(1) QLYREFN(1) SHVNVPS(1) TTANVRI(1) ESRVMSR(1) FSGGGNH(1) NQIYSAN(1) TTQVLEA(1) ETALIAA(1) TSQYLMI(1) 23 Phage display (common panning) 3 PMID:30592604 Gold Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The biopanning process was performed using the gold-patterned microfluidic chip with NEB Ph.D.-7 phage display peptide library kit. Ten microliters of phage solution (2e12 pfu mL−1) in 0.1 vol % PBST (PBS (pH 7.4) containing 0.1 vol % Tween 20) was loaded into the microfluidic biopanning platform at a flow rate of 1 μL min−1 at room temperature. Then PBST (0.1 vol % Tween 20 in the first round and 0.5 vol % Tween 20 from the second round) was introduced at 0.5 mL min−1 for 30 min to remove nonspecifically or weakly bound phages. The remaining high-affinity phages were eluted by injecting an elution buffer (0.2 M glycine-HCl (pH 2.2) with 1 mg mL−1 BSA) at 0.1 mL min−1 for 10 min, and the eluate was neutralized with 150 μL of 1 M Tris-HCl (pH 9.1). As the rounds progress, the proportion of high-affinity bacteriophages increases and fewer bacteriophages are removed from the surface after the washing step. Thus, the number of eluted bacteriophages increases as the rounds continue. 3399 TWLPLSL(1) VTWLPNI(1) TWVSGTV(1) SFAGTAF(1) ANDDRLV(1) ANPGVWL(1) SWTALGP(1) SHVNVPS(1) AHINVPS(1) VSPSYII(1) VVISDRY(1) EYWQDKP(1) LNKVAVP(1) DHITGYS(1) HTAITLT(1) YPQTFAT(1) HSGYIRF(1) TPFYGSW(1) HSHTLTW(1) NGAMVGV(1) HAAWHFI(1) QLAVAPS(1) ETALIAA(1) QLYREFN(1) 24 Phage display (common panning) 2 PMID:30592604 Gold Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The biopanning process was performed using the gold-patterned microfluidic chip with NEB Ph.D.-7 phage display peptide library kit. Ten microliters of phage solution (2e12 pfu mL−1) in 0.1 vol % PBST (PBS (pH 7.4) containing 0.1 vol % Tween 20) was loaded into the microfluidic biopanning platform at a flow rate of 1 μL min−1 at room temperature. Then PBST (0.1 vol % Tween 20 in the first round and 0.5 vol % Tween 20 from the second round) was introduced at 0.5 mL min−1 for 30 min to remove nonspecifically or weakly bound phages. The remaining high-affinity phages were eluted by injecting an elution buffer (0.2 M glycine-HCl (pH 2.2) with 1 mg mL−1 BSA) at 0.1 mL min−1 for 10 min, and the eluate was neutralized with 150 μL of 1 M Tris-HCl (pH 9.1). In the first round, a washing buffer containing a lower concentration of surfactant(0.1 vol % PBST) was used, which allows a higher diversity in the eluted phage pool and preserves more potential high-affinity bacteriophages.The surfactant concentration was then increased to 0.5 vol % PBST from the second round so that only phages having high affinity to the target remain. 3400 GVEGHKP(7) MVGKPQP(1) QTTSVGT(1) YNGSANQ(1) HGGVRLY(1) FSGGGNH(1) 6 Phage display (common panning) 5 PMID:30592604 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) A control experiment was also performed using another microfluidic device with the same initial phage display library(Ph.D.-7 phage display peptide library), which has no gold pattern on the surface, to exclude the possibility that the identified peptide was obtained by the interaction with PDMS, glass, or PEG coating. 3401 YCHPQFCG(4) HCHPQFCS(3) LCHPQFCG(2) DCHPQFCS(2) NCHPQFCP(1) DCHPQFCR(1) DCHPQFCV(1) HPQ 7 Phage display (common panning) 3 PMID:8508957 Streptavidin Biotin Not determined. Not determined. TN2 phage display library (XCX4CX) Library fractionation involved a 2-h incubation of phage with target, ten rapid washes with PBS containing 0.1% Tween-20 (to remove unbound phage), and stepwise washes with citrate buffers of pH 7, 6, and 5. Bound phage were removed from target-coated wells by the addition of pH 2 citrate-buffered elution solution for 10 min followed by immediate neutralization with Tris pH 8 buffer to a final concentration of 250 mM Tris. A portion of the phage eluted at pH 2 was used to grow a fresh stock of display phage which was then further fractionated. This sequence of events constitutes a single round of enrichment and amplification. 3402 LCYGGFCD(2) SCYGAFCR(1) VCYGAFCR(1) RCPTAGCD(1) NCYGAFCH(1) LCYGGFCP(1) 6 Phage display (common panning) 3 PMID:8508957 Anti-beta-endorphin monoclonal antibody 3-E7 Beta-endorphin Not determined. Not determined. TN2 phage display library (XCX4CX) Library fractionation involved a 2-h incubation of phage with target, ten rapid washes with PBS containing 0.1% Tween-20 (to remove unbound phage), and stepwise washes with citrate buffers of pH 7, 6, and 5. Bound phage were removed from target-coated wells by the addition of pH 2 citrate-buffered elution solution for 10 min followed by immediate neutralization with Tris pH 8 buffer to a final concentration of 250 mM Tris. A portion of the phage eluted at pH 2 was used to grow a fresh stock of display phage which was then further fractionated. This sequence of events constitutes a single round of enrichment and amplification. 3403 WTRSDHRIQ(2) WTRDQHQIH(1) WTLREHDFH(1) WQITQHKLQ(1) WTLQHHRVV(1) WTLGEHTLI(1) WRLSDHRMV(1) WTIKDHQLL(1) WKLSEHRMA(1) WSLGQHRIF(1) HGKHTHKVG(1) HGDKHKHRG(1) KPHQHKVHK(1) 13 Phage display (common panning) 3 PMID:10211822 Protein A Not determined. Not determined. Not determined. J404 phage display library (X9) Phage were selected by direct binding to the protein A matrix in the absence of antiserum for three rounds. 3404 LTFQGLP SHRGPSF GHWHFQE 3 Phage display (common panning) 3 PMID:11314263 Goat anti-human IgM polyclonal antibody Human immunoglobulin M (IgM) Not determined. Not determined. Ph.D.-7 phage display library (X7) 3405 GLTFQLHHQMRP LSFQNTVPPWAT GLMFQHPTLVPW KLNFQGSPYEPR LQFQNIESPLSP LQFQSLPDFTYS GLTFNNPAPFAG SLSFQTLKPLHP DHFSFQGLSSTG SHFSFQTGRSAL SHWSFQDRLSAI GHWGFQKDDPPW HSGDWLTNSPRV 13 Phage display (common panning) 3 PMID:11314263 Goat anti-human IgM polyclonal antibody Human immunoglobulin M (IgM) Not determined. Not determined. Ph.D.-12 phage display library (X12) 3406 GQSEKHL HGGVRLY SLSKWSF KIAVIST TDNTKPK TNWRTIN VSRDTPQ 7 Phage display (common panning) 3 PMID:30854940 Endothelial Progenitor Cell (EPC) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) 3407 SFKIPYHYDSGQ TIPRAPSPANTY NRPDSAQFWLHH 3 Phage display (subtractive panning) 3 PMID:30854940 Endothelial Progenitor Cell (EPC) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The positive selection was performed against Endothelial Progenitor Cells (EPCs) just after the negative selection against platelets, and the eluted phage clones were amplified only after these steps were completed. 3408 CPLDLRSPC[27.5 ± 0.5] 1 Phage display (common panning) 3 PMID:30867462 B1 peptide (CMSDWTGG) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Binding of the B1.12 peptide to its target B1 was evaluated by phage ELISA. Blocking buffer without B1 peptide was used as negative control. All experiments were done in triplicate and the binding of the phage clones to the B1 target was calculated according to the following formula: binding = OD phage/OD control × 100, where OD control and OD phage represent the OD410 nm values of phage binding with negative control and with B1 peptide, respectively. The binding value was regenerated from Figure 1a and shown. A solution of 1 mg/mL of the B1 peptide in 100 mM NaHCO3 (pH 8.6) was prepared and 100 μL was coated on a 96-well plate and incubated overnight at 4 °C. The coating solution was poured off and blocking buffer (skimmed milk at 5 mg/mL in 100 mM NaHCO3) was added for 1 hour at 4 °C. After washing with 0.1% (v/v) Tween-20/TBS, 10 μL (2e11 plaque forming units, pfu) of original library, Ph.D™-C7C Phage Display Peptide (New England Biolabs), were diluted in 100 μL TBST and added to the plate well for 2 hours at room temperature with gentle agitation. After ten washes with 0.1% TBST, the bound phages were eluted with 0.2 M glycine-HCl (pH 2.2) and neutralized with 1 M Tris-HCl (pH 9.1). Eluted phages were amplified in 20 mL LB inoculated with the E. coli ER2738 strain, purified by PEG/NaCl precipitation, and titrated as described in the NEB standard protocol to be used for the next round of selection. The two next rounds of selection were performed under more stringent conditions as the concentration of Tween-20 was gradually increased (0.3% and 0.5% for the second and third round, respectively) and the incubation time of phages with the target was gradually reduced (1.5 hours and 1 hour in the second and third rounds, respectively). We identified by Phage display a novel peptide called B1.12 (ACPLDLRSPCG) which selectively binds to the extracellular loop (B1) of the latent membrane protein 1 (LMP1) as demonstrated by molecular docking, NMR and ITC. Using an LMP1 expressing cell line, we showed that B1.12 decreased cell viability, and induced G0/G1 cell cycle arrest. In addition, the expression of A20, pAkt, and pNFkb (pRelA536) in C666-1 cells treated with B1.12 decreased compared to the untreated cells. In conclusion, we selected a novel peptide able to bind specifically to the extracellular loop of LMP1 and thus modulate its oncogenic properties. 3409 GTTVSQFLS(2) VSPSLINEV(1) SVPTLVNLT(1) VVPVVVRTS(1) VAPVVVVLF(1) VSPVVVVEV(1) NFPVVVWDV(1) WAPLVVMDW(1) GVMPEVVTE(1) VEVVQLHVS(1) VEGVIILGS(1) LEPVTISAG(1) LLEPILEKR(1) GPPVLMGILI(1) FRLVTLSLG(1) ARTVWGFAG(1) VVQTVMLIS(1) TVQVLERLA(1) MVVASDVIG(1) EVLVVDKVM(1) SVVLATLKI(1) AVTLAQLRL(1) VMLAYLAVP(1) GVLVSLATS(1) GLTVADSGDL(1) GLYIAESFY(1) GLVIRELRRL(1) GLMVRETLGT(1) GLLISLVAR(1) GLLVSEVWTA(1) VGTLLVSYM(1) GALTVSLRV(1) GALLVFERW(1) WVLAVMEHR(1) ECLVIEELR(1) CRLVVIEHV(1) KLVINELVR(1) VAIRVMVRC(1) FTKIREVLG(1) 39 Phage display (common panning) 5 PMID:12900423 Chymase (EC:3.4.21.39) Not determined. Not determined. Not determined. X9 T7 phage display library An aliquot of the amplified phages (~e9 plaque forming units) were allowed to bind to 100 μl of Ni-NTA-agarose beads for 1 h while rotating gently. Unbound phages were removed by washing 5 times with 1.5 ml of 1 M NaCl, 0.1% Tween 20 in PBS (pH 7.2) and further washed twice in 1.5 ml of PBS and finally resuspended in 270 μl of PBS. A control elution of the phages with 100 μl of 500 mM imidazole concluded that at least 1.0e8 phages were attached to the matrix after washing. rMCP-5 was digested with enterokinase as previously described in the absence of heparin and then subjected to heparin-Sepharose purification. After elution with PBS containing 1.5 M NaCl, heparin was added to a 10:1 mass ratio of heparin to protease. The selection was started by adding ~0.2 μg of protease (~27 nM final concentration) or buffer without protease as a control to the tubes with the resuspended beads. The protease was allowed to digest susceptible phages overnight with gentle agitation. To recover released phages, the supernatant was removed, and the beads were washed in 100 μl of PBS. To ensure that all recovered phages lacked the histidine tag, 15 μl of fresh Ni-NTA-agarose beads were added to the phage suspension, and the mixture was agitated for 15 min followed by centrifugation to recover the supernatant. Ten μl of the supernatant was used to determine the amount of detached phages in each round of selection. The remaining 290 μl of the supernatant was added to a 10-ml culture (A = ~0.6) of Escherichia coli (BLT5615). The bacteria had been induced with 100 μl of 100 mM isopropyl-1-thio-β-D-galactopyranoside 30 min before phage addition to ensure production of the phage capsid protein. After six rounds of selection, 40 plaques were arbitrarily isolated from LB plates. The oligonucleotide inserted into the phage DNA was amplified by PCR, and the randomized sequence was determined by nucleic acid sequencing. 3410 LVTFLSLSA GILFLSSLIS LAVLFRSLL STLFVGSSV VALLWSLVS VMLWVSSAA GQLYSSVLLD VLQRFSMER VMRFVMAIG VVRFLSGLC IVRWLSINV IMTWISVET RCSAFVLES VLLFMGNWV VALFWLQQV VSLFMGIDF VQLYWVVEE SLELWMSMW GVELFFLRM ELRMLFAAF GGLYSYVVME VVVYGGFEI IVVVRYSLF VERWSWVLV WGVSFLSVV DVWFNLLLN EISMYFSLV VRCFMALGY VMMSYTFVRH VPMAWIGYA WEPDRWVSFH GVRFLSFENW MLLRFWAYE WVVWISKEF WVSYTSRLW WWAVVSFVGH YVTFGGLWG SLMFFWARL WDWMPFLAS GWMFYDAILT 40 Phage display (common panning) 5 PMID:17681377 Chymase (EC:3.4.21.39) Not determined. Not determined. Not determined. X9 T7 phage display library An aliquot of the amplified phages (~e9 pfu) was allowed to bind to 100 μl Ni-NTA agarose beads for 1 h while rotating gently at 4 ◦C. Unbound phages were removed by washing ten times with 1.5 ml 1 M NaCl, 0.1% Tween-20 in PBS, pH 7.2, and two subsequent washes with 1.5 ml PBS. The beads were finally resuspended in 1ml PBS. Activated and heparin-sepharose purified mast cell protease 1 was added to the resuspended beads to start the digestion of susceptible peptides. Buffer without protease was used as a control. Protease digestion was allowed to proceed at room temperature over night with gentle agitation. To recover released phages, the Ni-NTA agarose beads were pelleted by quick centrifugation and the supernatant containing the phages was removed. To ensure that all of the released phages were recovered the beads were resuspended in 100 μl PBS (pH 7.2), which was removed and added to the first supernatant after centrifugation. A control elution of the remaining phages still bound to the Ni-NTA agarose beads, using 100μl 100mM imidazole concluded that at least 1.0e8 phages were attached to the matrix during each selection round. To ensure that all phages recovered after the protease digestion step lacked the His6 tag, 15μl fresh Ni-NTA agarose beads were added to the phage suspension and the mixture agitated for 5 min followed by centrifugation to recover the supernatant. Thirty microlitres of the supernatant was used to determine the amount of detached phages in each round of selection. The remaining 1070 μl of the supernatant was added to a 10ml culture (OD~0.6) of E. coli BLT5615. The bacteria had previously been induced to produce the phage capsid protein by adding 100 μl 100m MIPTG 30 min prior to phage addition. Approximately 75 min after phage addition the bacteria lysed. The lysate was centrifuged to remove cell debris and 900 μl of the phage sub-library was added to fresh agarose beads. After binding and washing the sub-library, a new round of selection was started. Following five rounds of selection, ~100 plaques per enzyme were arbitrarily isolated from LB plates and each dissolved in phage extraction buffer (100mM NaCl and 6mM MgSO4 in 20mM Tris–HCl pH 8.0) and vigorously shaken for 30 min in order to disrupt the phages. 3411 GVLFVEASA VSLFLEVQE GVLFLELEGV GGAVLFLEL VVLWSDIGE SVMRWSEVL PVAYAAGGV HMMVVYRNS EVVLRSDWNHH WLAYSEEEV SRFVSFVDD LVSYGEVFL RVLQYLEYSP IAVFLDVYE VARFLDLSF WEVWMEMRT GWDLWQDIS WDVPMLWRDH VVSVFYNQD ISLYLAPIW PFSNFMSLG WVTTYLSGG GWMLRYLAG FLSFWDLAG TYFSFWDLAG TYSVTFTEH WGWLLFREL AWALYVEYG WEAVGYMGW WQVLRFWGM WASVQFFMG WAPWTYLLT IWMFYFSSA GCERFFRWI GGWWLTAYISI WVSLWFWEE WVVYGGWYL WYYFSSYIF PGGWDWGYYVGY 39 Phage display (common panning) 5 PMID:17681377 Mast cell protease 4 (EC:3.4.21.-), mMCP-4 Not determined. Not determined. Not determined. X9 T7 phage display library An aliquot of the amplified phages (~e9 pfu) was allowed to bind to 100 μl Ni-NTA agarose beads for 1 h while rotating gently at 4 ◦C. Unbound phages were removed by washing ten times with 1.5 ml 1 M NaCl, 0.1% Tween-20 in PBS, pH 7.2, and two subsequent washes with 1.5 ml PBS. The beads were finally resuspended in 1ml PBS. Activated and heparin-sepharose purified mast cell protease 4 was added to the resuspended beads to start the digestion of susceptible peptides. Buffer without protease was used as a control. Protease digestion was allowed to proceed at room temperature over night with gentle agitation. To recover released phages, the Ni-NTA agarose beads were pelleted by quick centrifugation and the supernatant containing the phages was removed. To ensure that all of the released phages were recovered the beads were resuspended in 100 μl PBS (pH 7.2), which was removed and added to the first supernatant after centrifugation. A control elution of the remaining phages still bound to the Ni-NTA agarose beads, using 100μl 100mM imidazole concluded that at least 1.0e8 phages were attached to the matrix during each selection round. To ensure that all phages recovered after the protease digestion step lacked the His6 tag, 15μl fresh Ni-NTA agarose beads were added to the phage suspension and the mixture agitated for 5 min followed by centrifugation to recover the supernatant. Thirty microlitres of the supernatant was used to determine the amount of detached phages in each round of selection. The remaining 1070 μl of the supernatant was added to a 10ml culture (OD~0.6) of E. coli BLT5615. The bacteria had previously been induced to produce the phage capsid protein by adding 100 μl 100m MIPTG 30 min prior to phage addition. Approximately 75 min after phage addition the bacteria lysed. The lysate was centrifuged to remove cell debris and 900 μl of the phage sub-library was added to fresh agarose beads. After binding and washing the sub-library, a new round of selection was started. Following five rounds of selection, ~100 plaques per enzyme were arbitrarily isolated from LB plates and each dissolved in phage extraction buffer (100mM NaCl and 6mM MgSO4 in 20mM Tris–HCl pH 8.0) and vigorously shaken for 30 min in order to disrupt the phages. 3412 TSVLFSLRM GTPFSLRARG PSLYSLSRV IVRYSLAGE VAMFSVRVS LANFSVRIA EVVRFSVSS LVIGYSARP ASVTLFSTR VVSLFRLGV PVVLFRLSV SPVLFRVRS PVVGQIFRVH RIATMFRVE PGGYEVLTLS RGQWPRTVA KSWYSLSSE DWSWFSLSM YFRYFSLGG WQWYSLGLH GTWFSLGVMD GGWYSVALADK MTWFSSQVA VFWRFFSNA GLFFSMYTRV VVWFRSMAD QPVFFRIRG DFQMFFRLS YSTRFFTLG GGFYTVANFRG WSFFALSRQ WNLSKFSLS SRLWRTFSLH PVLYSLGYV PGGYSVRVFNSS TLIMTFSAG GIVRFSARW GWRRVLFSAH YLTVNFSTS VRVQMFRFSHH PGGYRAMKNFAM WRTYRSAVV YMQASFALG GYQAARFAAH 44 Phage display (common panning) 5 PMID:18313755 Chymase (EC:3.4.21.39) Not determined. Not determined. Not determined. X9 T7 phage display library A 300 μl aliquot of the amplified phages (~1.0e9 pfu) was allowed to bind to 100 μl nickel-nitrilotriacetic acid (Ni-NTA) agarose beads for 1 h while rotating gently. Unbound phages were removed by washing 15 times with 1.5 ml 1M NaCl, 0.1% Tween-20 in PBS (pH 7.2), and further washed twice in 1.5 ml PBS (pH 7.2) and finally resuspended in 1ml PBS (pH 7.2). To determine the amount of phages bound to the matrix, a control elution of the phages with 100 μl 100mM imidazole was performed. This analysis showed that at least 1.0e8 phages were attached to the matrix after washing. Purified mMCP-1 (0.6 μg) was added to the tubes with the beads resuspended in PBS (pH 7.2), to start the selections. PBS (pH 7.2) without protease was used as control. The proteases were allowed to digest susceptible phages at room temperature over night under gentle agitation. To recover released phages, the Ni–NTA agarose beads were pelleted by centrifugation and the phages in the supernatant were removed in a total volume of 1000 + 100 μl PBS (pH 7.2). To ensure that all phages recovered lacked the histidine tag, 15 μl fresh Ni–NTA agarose beads were added to the phage suspension and the mixture agitated for 15 min followed by centrifugation. The supernatant was recovered and transferred to a new tube. 10 μl of the supernatant was used to determine the amount of detached phages in each round of selection. The remaining 1090 μl of the supernatant was added to a 10ml culture (OD ~0.6) of E. coli (BLT5615). The bacteria had been induced with 100 μl 100mM IPTG 30 min before phage addition, to ensure production of the phage capsid protein. Approximately 75 min after phage addition the bacteria lysed and 900 μl of the phage sublibrary was added to fresh agarose beads. After binding and washing the sub-library, a new round of selection was started. Following five rounds of selection, phages were plated on LB plates supplemented with ampicilin. Plaques corresponding to single phage clones were isolated and dissolved in phage extraction buffer (100mM NaCl, 20mM Tris–HCl pH 8.0 and 6mM MgSO4) and vigorously shaken for 30 min in order to extract the phages from the agarose. 3413 IAVFWEGTP(2) ALEVYASGG(1) AAVIAMFAGH(1) LISSYGLEL(1) SMVSYADVS(1) GQVFAEVRLQ(1) LVTRFSEEE(1) LAEYSDPAV(1) LVSVAYSET(1) GLVAYSEVE(1) GALLYTDRV(1) GILYVDSSTT(1) GVLYEESGSR(1) SVLLFEDGL(1) CDNYEDIRR(1) VASFDSAVL(1) ARRVTFDSL(1) RSTFQAALL(1) RMTFSLSTL(1) VWVVEMFTTH(1) VFDRVVFAGH(1) WTREAWALV(1) WNPQAYGLT(1) VATTFFGLT(1) HLMSFAEEW(1) LVVYGDYTT(1) GVRYFDALL(1) GGEWYEASQGG(1) WRLCPFMDA(1) AVWLYSERL(1) AWLAYTDSV(1) FLAVEAFMDH(1) WDAYNEAVE(1) GEFYSEGLGH(1) AIWWSEVVR(1) GLLYSDVGIW(1) AVSFSFTTL(1) GQAYSMALPY(1) GVLWSAFARG(1) LLLFSGGFL(1) WFRVAVYGSH(1) QVAFFGLYD(1) PGLLAFFLF(1) YVGSSVYFGH(1) LWSVYFEPM(1) FFISVYVEA(1) GFTFFDVGSY(1) [GLV]-[VAL]-[AVL]-[YF]-S-[DE]-[AVG] 47 Phage display (common panning) 5 PMID:19073880 Chymase (EC:3.4.21.39) Not determined. Not determined. Not determined. X9 T7 phage display library An aliquot of the amplified phages (~1.0e9 pfu) was allowed to bind to 100 μl Ni–NTA agarose beads for 1 h while rotating gently at 4 ◦C. Unbound phages were removed by washing 10 times with 1.5 ml 1 M NaCl, 0.1% Tween 20 in PBS, pH 7.2, and two subsequent washes with 1.5 ml PBS. The beads were finally re-suspended in 1000 μl PBS. Activated and heparin–sepharose-purified protease (~0.1 μg) was added to the re-suspended beads to start the selection of susceptible peptides. Buffer without protease was used as a control. Protease digestion was allowed to proceed at room temperature overnight with gentle agitation. To recover released phages, the Ni–NTA agarose beads were pelleted by centrifugation in a table top centrifuge and the supernatant containing the phages was removed. To ensure that all the released phages were recovered, the beads were re-suspended in 100 μl PBS (pH 7.2), and the supernatant after mixing and centrifugation was added to the first recovered phages. A control elution of the phages still bound to the beads, using 100 μl 100 mM imidazole, concluded that at least 1.0e8 phages were attached to the matrix during each selection. To ensure that the His6 tag had been hydrolyzed on all phages recovered after protease digestion, 15 μl fresh Ni–NTA agarose beads were added to the phage suspension and the mixture agitated for 15 min followed by centrifugation to recover the supernatant. Thirty microliters of the supernatant was used to determine the amount of phages detached in each round of selection. The remaining 1070 μl of the supernatant was added to a 10-ml culture (optical density ~0.6) of Escherichia coli (BLT5615). The bacteria had previously been induced to produce the T7 phage capsid protein by adding 100 μl 100 mM isopropyl β-D-1-thiogalactopyranoside 30 min prior to phage addition. The bacteria lysed ~75 min after phage addition. The lysate was centrifuged to remove ceμl debris and 900 μl of the phage sublibrary was added to 100 μl fresh agarose beads. After binding and washing the sublibrary, a new round of selection was started. Following five rounds of selection, 50 plaques were arbitrarily isolated from luria broth plates and each dissolved in phage extraction buffer (100 mM NaCl and 6 mM MgSO4 in 20 mM Tris–HCl, pH 8.0) and vigorously shaken for 30 min in order to extract the phages from the agarose. 3414 QRKLAAKLT 1 Phage display (common panning) 6 PMID:18068970 Pseudomonas aeruginosa ATCC 27853 Not determined. Not determined. Not determined. pVIII-9aa phage display library (X9) 3415 RVRSWRVRR(2) LVRPRAGRS(1) PRALRSVVR(1) PRAMRGGRR(1) PRGMRVVQR(1) TPRSARFGD(1) TLMVPRTGS(1) PRSWWKAAW(1) LVYMPRAFL(1) RSARFKSIW(1) RAAKSGLHF(1) RAFRWGRAP(1) RSRSSRLKV(1) RSSRVTSRM(1) SLRIRSSRS(1) LRLKLDIGR(1) RRQCRALRY(1) 17 Phage display (common panning) 5 PMID:22384068 Prothrombin Not determined. Not determined. Not determined. X9 T7 phage display library In brief, phages are anchored to nickel nitrilotriacetic acid (Ni-NTA) beads via the His6-tags before the first protease treatment. The protease is added and allowed to react over night, releasing phages displaying cleavage-susceptible 9-mers from the beads. Samples are centrifuged and cleaved phages are collected in the supernatant. These phages are amplified in the E. coli strain BLT5615 and enter the next selection round (biopanning). After five biopannings with 1 U of thrombin, enriched cleavage-susceptible phages are sequenced. 3416 LMPRTKRRG(2) LSPRSVRLL(1) GLMPRGVRL(1) VRARSRNSV(1) SRSKGRARN(1) RRRSSVEPF(1) PRAFKVANL(1) GGPRSWRRG(1) ARPRSWLVM(1) GFKPRTFYV(1) LVPRAYYYW(1) WGRFQGWSW(1) WTFATGPFM(1) TLMVPRTGS(1) RSVRGGRLW(1) VMRAGRWLW(1) 16 Phage display (common panning) 5 PMID:22384068 Prothrombin Not determined. Not determined. Not determined. X9 T7 phage display library In brief, phages are anchored to nickel nitrilotriacetic acid (Ni-NTA) beads via the His6-tags before the first protease treatment. The protease is added and allowed to react over night, releasing phages displaying cleavage-susceptible 9-mers from the beads. Samples are centrifuged and cleaved phages are collected in the supernatant. These phages are amplified in the E. coli strain BLT5615 and enter the next selection round (biopanning). After five biopannings with 0.2 U of thrombin, enriched cleavage-susceptible phages are sequenced. 3417 PCAIWF(58%) WVCSGG(42%) 2 Phage display (common panning) 3 PMID:27819042 Extracellular domain of vascular endothelial growth factor receptor 3 (EC:2.7.10.1) (VEGFR-3) Vascular endothelial growth factor C and D Not determined. Not determined. fUSE55-based X6 phage display library To isolate peptides targeting VEGFR-3, microtiter wells were coated with recombinant mouse VEGFR-3/Fc [1 mg in 50 ml of phosphatebuffered saline (PBS)] [10 mM Na2HPO4, 1.8 mM KH2PO4, 137 mM NaCl, and 2.7 mM KCl (pH 7.4)], blocked with PBS supplemented with 3% bovine serum albumin (BSA), and incubated with the X6 phage peptide library (e9 TU) in 50 ml of PBS supplemented with 1% BSA (PBS/BSA) for 2 hours at room temperature. The wells were then washed, and bound phages were recovered by bacterial infection and amplified for the next cycle of selection. After the third round of selection, random bacterial colonies were selected for DNA sequencing to identify the phage coding peptides. These peptides (PCAIWF and WVCSGG) specifically prevent the binding of vascular endothelial growth factor (VEGF) family members to all three receptors and downstream signaling but do not affect other angiogenic receptor tyrosine kinases (RTKs) and their ligands. One of the selected peptides (PCAIWF) was also very effective at preventing pathological angiogenesis in a mouse model of retinopathy, normalizing the vasculature to levels similar to those of a normal developing retina. Collectively, our results suggest that these peptides are pan-VEGF inhibitors directed at a common binding pocket shared by all three VEGFRs. These peptides and the druggable binding site they target might be important for the development of novel and selective smallmolecule, extracellular ligand-binding inhibitors of RTKs (eTKIs) for angiogenic-dependent diseases. 3418 LPPWKLK(11/27) WSLSELH(4/27) LHRHANL(2/27) IGASVHR(1/27) AHHLKVS(1/27) NHPLYNR(1/27) ALEHTSR(1/27) HPAIRPP(1/27) SSNQFHQ(1/27) KVPGHQQ(1/27) TLAPRTA(1/27) 11 Phage display (common panning) 4 PMID:30889924 Polycrystalline SnO powder Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) For each biopanning procedure, 20 mg of the SnO powder were equilibrated for 10 min in 1 mL Tris-buffered saline + 0.1 % Tween-20 (TBST0.1) in an ultrasonic bath. After centrifugation at 10621 g for 2 min followed by removal of the supernatant, the SnO powder was resuspended in 200 μL TBST0.1. 10 μL of the phage peptide library (~1.2e11 phages) were added for 1 h under light agitation at 20 °C. To remove unbound and weakly bound phages, the SnO powder was washed 10 times with 1 mL TBST0.5. For this, first the powder was sedimented by centrifugation at 10621 g for 2 min. Subsequently, the supernatant was removed and the powder resuspended in 1 mL TBST0.5 by gentle vortexing, followed by another centrifugation step. Bound phages were eluted by 1 mL of 0.2 M Glycine-HCl (pH 2.2) supplemented with 1 mg/mL BSA. After 10 min of gentle mixing, the powder is sedimented by centrifugation, and the supernatant is transferred to a new 1.5 mL tube and neutralized by adding 150 μL of 1 M Tris–HCl (pH 9.1). The obtained M13 phages were amplified in E. coli ER2738, according to manufacturer’s recommendations. 3419 LPPWKLK(17/23) WSLSELH(2/23) LHRHANL(1/23) SSNQFHQ(1/23) VGKTHAD(1/23) FPLHELR(1/23) 6 Phage display (common panning) 5 PMID:30889924 Polycrystalline SnO powder Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) For each biopanning procedure, 20 mg of the SnO powder were equilibrated for 10 min in 1 mL Tris-buffered saline + 0.1 % Tween-20 (TBST0.1) in an ultrasonic bath. After centrifugation at 10621 g for 2 min followed by removal of the supernatant, the SnO powder was resuspended in 200 μL TBST0.1. 10 μL of the phage peptide library (~1.2e11 phages) were added for 1 h under light agitation at 20 °C. To remove unbound and weakly bound phages, the SnO powder was washed 10 times with 1 mL TBST0.5. For this, first the powder was sedimented by centrifugation at 10621 g for 2 min. Subsequently, the supernatant was removed and the powder resuspended in 1 mL TBST0.5 by gentle vortexing, followed by another centrifugation step. Bound phages were eluted by 1 mL of 0.2 M Glycine-HCl (pH 2.2) supplemented with 1 mg/mL BSA. After 10 min of gentle mixing, the powder is sedimented by centrifugation, and the supernatant is transferred to a new 1.5 mL tube and neutralized by adding 150 μL of 1 M Tris–HCl (pH 9.1). The obtained M13 phages were amplified in E. coli ER2738, according to manufacturer’s recommendations. 3420 LPPWKLK(11/27) WSLSELH(4/27) LHRHANL(2/27) IGASVHR(1/27) AHHLKVS(1/27) NHPLYNR(1/27) ALEHTSR(1/27) HPAIRPP(1/27) SSNQFHQ(1/27) KVPGHQQ(1/27) TLAPRTA(1/27) FRW 11 Phage display (in vivo) 3 PMID:30670660 Olfactory bulk Not determined. Not determined. Not determined. fUSE55-based CX8C phage display library (CX8C) To isolate peptides targeting the brain, animals received the CX8C phage library by i.v. injection. After 30 minutes in circulation, mice were perfused with 20 mL of Dulbecco’s modified Eagle’s medium (DMEM) and different regions of the brain (cerebellum, olfactory bulb and hemispheres) were collected. Phage bound to tissue were recovered by tissue homogenization followed by bacterial infection, transferred to LB media supplemented with kanamycin (100 μg/ml) and tetracylcin (20 μg/ml) amplified by overnight culture and purified from culture supernatants by the PEG/NaCl method. Two more successive rounds of selection were then performed by reinjecting i.v. the recovered pools of phage into different mice (one for each brain area). Peptides containing a conserved motif in which amino acids Phenylalanine−Arginine−Tryptophan (FRW) predominate could be visualized by transmission electron microscopy bound to the junctions between endothelial cells in all areas of the brain, including the optic nerve, but not in other barrier-containing tissues, such as intestines and testis. Remarkably, peptides containing the motif do not bind to vessels in the retina, implying an important molecular difference between these two vascular barriers. Furthermore, the peptide allows for in vivo imaging, demonstrating that new tools for studying and imaging the brain are likely to emerge from this motif. 3421 CLYVNFAWRC(10) CVWANFRWQC(6) CFFADFRWYC(5) CFFVFPRWYC(2) CYSFLTSDFC(1) CFFDAIEFRC(1) CGLAVGTADC(1) CYLNWRWSVC(1) CSVNLVFGSC(1) CVYMYFGWFC(1) CMWVAWRWVC(1) CYFYNYRWWC(1) CVWMNYRWVC(1) FRW 13 Phage display (in vivo) 3 PMID:30670660 Hemisphere Not determined. Not determined. Not determined. fUSE55-based CX8C phage display library (CX8C) To isolate peptides targeting the brain, animals received the CX8C phage library by i.v. injection. After 30 minutes in circulation, mice were perfused with 20 mL of Dulbecco’s modified Eagle’s medium (DMEM) and different regions of the brain (cerebellum, olfactory bulb and hemispheres) were collected. Phage bound to tissue were recovered by tissue homogenization followed by bacterial infection, transferred to LB media supplemented with kanamycin (100 μg/ml) and tetracylcin (20 μg/ml) amplified by overnight culture and purified from culture supernatants by the PEG/NaCl method. Two more successive rounds of selection were then performed by reinjecting i.v. the recovered pools of phage into different mice (one for each brain area). Peptides containing a conserved motif in which amino acids Phenylalanine−Arginine−Tryptophan (FRW) predominate could be visualized by transmission electron microscopy bound to the junctions between endothelial cells in all areas of the brain, including the optic nerve, but not in other barrier-containing tissues, such as intestines and testis. Remarkably, peptides containing the motif do not bind to vessels in the retina, implying an important molecular difference between these two vascular barriers. Furthermore, the peptide allows for in vivo imaging, demonstrating that new tools for studying and imaging the brain are likely to emerge from this motif. 3422 CIWVDFRWKC(4) CVWVDFHWVC(2) CVWVSFHWVC(2) CFWWKYVYRC(1) CVYHGFRWRC(1) CRYIDFRWSC(1) CFWYGMRWWC(1) CFWYSYRWIC(1) CRYSAWKWWC(1) CWRALNYSPC(1) CGSDGEVGRC(1) FRW 11 Phage display (in vivo) 3 PMID:30670660 Cerebellum Not determined. Not determined. Not determined. fUSE55-based CX8C phage display library (CX8C) To isolate peptides targeting the brain, animals received the CX8C phage library by i.v. injection. After 30 minutes in circulation, mice were perfused with 20 mL of Dulbecco’s modified Eagle’s medium (DMEM) and different regions of the brain (cerebellum, olfactory bulb and hemispheres) were collected. Phage bound to tissue were recovered by tissue homogenization followed by bacterial infection, transferred to LB media supplemented with kanamycin (100 μg/ml) and tetracylcin (20 μg/ml) amplified by overnight culture and purified from culture supernatants by the PEG/NaCl method. Two more successive rounds of selection were then performed by reinjecting i.v. the recovered pools of phage into different mice (one for each brain area). Peptides containing a conserved motif in which amino acids Phenylalanine−Arginine−Tryptophan (FRW) predominate could be visualized by transmission electron microscopy bound to the junctions between endothelial cells in all areas of the brain, including the optic nerve, but not in other barrier-containing tissues, such as intestines and testis. Remarkably, peptides containing the motif do not bind to vessels in the retina, implying an important molecular difference between these two vascular barriers. Furthermore, the peptide allows for in vivo imaging, demonstrating that new tools for studying and imaging the brain are likely to emerge from this motif. 3423 CSWYSYRWIC CMYHNFAWYC CYYAFFRWHC CIYSFFAWRC CFFWMFRWIC CSWFDFRWHC CVYLDFKWQC CVWVSYRWVC CIYFQYHWVC CTWVNFRYVC [FYW]-X-X-[FYW]-[ARKH]-[FYW] 1021 Phage display (in vivo) 3 PMID:30670660 Brain Not determined. Not determined. Not determined. fUSE55-based CX8C phage display library (CX8C) To isolate peptides targeting the brain, animals received the CX8C phage library by i.v. injection. After 30 minutes in circulation, mice were perfused with 20 mL of Dulbecco’s modified Eagle’s medium (DMEM) and different regions of the brain (cerebellum, olfactory bulb and hemispheres) were collected. Phage bound to tissue were recovered by tissue homogenization followed by bacterial infection, transferred to LB media supplemented with kanamycin (100 μg/ml) and tetracylcin (20 μg/ml) amplified by overnight culture and purified from culture supernatants by the PEG/NaCl method. Two more successive rounds of selection were then performed by reinjecting i.v. the recovered pools of phage into different mice (one for each brain area). Peptides containing a conserved motif in which amino acids Phenylalanine−Arginine−Tryptophan (FRW) predominate could be visualized by transmission electron microscopy bound to the junctions between endothelial cells in all areas of the brain, including the optic nerve, but not in other barrier-containing tissues, such as intestines and testis. Remarkably, peptides containing the motif do not bind to vessels in the retina, implying an important molecular difference between these two vascular barriers. Furthermore, the peptide allows for in vivo imaging, demonstrating that new tools for studying and imaging the brain are likely to emerge from this motif. 3424 CNARGDMHC(3) CIVRGDNVC(3) CTLRGDHHC(2) CTIRGDTHC(2) CNARGDAPC(2) CVPRGDMHC(2) CLRGDYPKC(1) CECRGDCYC(1) CRGDNTDQC(1) CVSRGDQIC(1) CVNRGDHSC(1) CNGQNWRGC(1) CLNGAYPNC(1) CSLKGDLKC(1) CSSSQSPVC(1) CHNNHYRDC(1) CHSTQMNHG(1) CFGSKAGQC(1) CHQTINRLC(1) CDRADKQGC(1) CTAATSTMC(1) CQNPNQKFC(1) RGD 22 Phage display (in vivo) 2 PMID:30730145 Blood Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Briefly, Ph.D.-C7C phage library (1.0e11 plaque forming units, or pfu) in 300 μL saline was injected into the rat through the tail vein. Forty-eight hours later, as much blood as possible was collected from the heart and plated out on LB plates containing X-gal (5-bromo-4-chloro- 3-indolyl-b-Dgalactoside) and IPTG (isopropyl-b-D-thiogalactoside). About 300 blue plaques, obtained from six rats, were picked, mixed and amplified. After purification, 1.0e11 of the amplified phage was used for second round biopanning, following the same procedure as for the first round screening but with blood withdrawn at 96 h post injection. About 500 blue plaques obtained from the second round screening were mixed, amplified, and used for the third round screening, with blood withdrawn at 120 h post injection. Thirty blue plaques each from the second and third round screening were randomly picked and subjected to DNA sequencing. The majority of the identified blood circulation-prolonging (BCP) peptides contained an arginine-glycine-aspartic acid (RGD) motif, which was necessary but insufficient for the circulation-prolonging activity. We further demonstrated that the RGD-mediated specific binding to platelets was primarily responsible for the enhanced blood retention of BCP1 (CNARGDMHC). The utility of the BCP1 peptide was demonstrated by fusion of the peptide to human heavy-chain ferritin (HFn), leading to significantly improved pharmacokinetic profile, enhanced tumor cell uptake and optimum anticancer efficacy for doxorubicin encapsulated in the HFn nanocage. Our results provided a proof-of-concept for an innovative yet simple strategy, which utilizes phage display to discover novel peptides with the capability of substantially prolonging blood circulation for engineered theranostic nanoparticles. 3425 CNARGDMHC(6) CIVRGDNVC(4) CVPRGDMHC(4) CLRGDYPKC(2) CECRGDCYC(2) CTIRGDTHC(2) CNARGDAPC(1) CRGDNTDQC(1) CPDATGGLC(1) CSNIMNNFC(1) CSTGWPRAC(1) CDNPDRRKC(1) CAIPNQVPC(1) CEMTNRSHC(1) CSNYMTTNC(1) CSGMYMAQC(1) 16 Phage display (in vivo) 3 PMID:30730145 Blood Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Briefly, Ph.D.-C7C phage library (1.0e11 plaque forming units, or pfu) in 300 μL saline was injected into the rat through the tail vein. Forty-eight hours later, as much blood as possible was collected from the heart and plated out on LB plates containing X-gal (5-bromo-4-chloro- 3-indolyl-b-Dgalactoside) and IPTG (isopropyl-b-D-thiogalactoside). About 300 blue plaques, obtained from six rats, were picked, mixed and amplified. After purification, 1.0e11 of the amplified phage was used for second round biopanning, following the same procedure as for the first round screening but with blood withdrawn at 96 h post injection. About 500 blue plaques obtained from the second round screening were mixed, amplified, and used for the third round screening, with blood withdrawn at 120 h post injection. Thirty blue plaques each from the second and third round screening were randomly picked and subjected to DNA sequencing. The majority of the identified blood circulation-prolonging (BCP) peptides contained an arginine-glycine-aspartic acid (RGD) motif, which was necessary but insufficient for the circulation-prolonging activity. We further demonstrated that the RGD-mediated specific binding to platelets was primarily responsible for the enhanced blood retention of BCP1 (CNARGDMHC). The utility of the BCP1 peptide was demonstrated by fusion of the peptide to human heavy-chain ferritin (HFn), leading to significantly improved pharmacokinetic profile, enhanced tumor cell uptake and optimum anticancer efficacy for doxorubicin encapsulated in the HFn nanocage. Our results provided a proof-of-concept for an innovative yet simple strategy, which utilizes phage display to discover novel peptides with the capability of substantially prolonging blood circulation for engineered theranostic nanoparticles. 3426 GWVSNTTQAHHV(9) AMDIAYRTHREP(3) DLNKPKPLYQQH(2) ESMTYLSTAPEK(2) HNYPVLRPNQIT(2) APNAPTQTTTPV(1) ARHPDTNYSYGA(1) MMKQTDQLLRNN(1) NWHVYSTISNQT(1) TRTPPESYASVR(1) YSGKDLPPMKDS(1) 11 Phage display (competitive panning) 3 PMID:30705359 Chondroitin-4-sulfate, C4S Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In each round of panning, binding phages were eluted with 100 μl of C4S (100 mg/ml in PBS) and applied to the next panning round. From the phage clones binding to C4S we selected three peptides (AMDIAYRTHREP, GWVSNTTQAHHV and TRTPPESYASVR) for further analysis. We observed that these peptides bind to C4S, but not chondroitin-6-sulfate, heparin sulfate or dermatan sulfate, in a concentration-dependent and saturable manner, whereas the scrambled peptides showed highly reduced or no binding to C4S. The C4S-binding peptides, but not their scrambled counterparts, when added to cultures of mouse cerebellar neurons and human neuroblastoma cells, neutralized the inhibitory functions of the C4S- and CSPG-coated substrate on cell adhesion, neuronal migration and neurite outgrowth. These results indicate that the C4S-binding peptides neutralize several inhibitory functions of CSPGs, suggesting that they may be beneficial in repairing mammalian nervous system injuries. 3427 KHSHLGSFERHL(3/24) HSPQYWVHHWRG(2/24) SPHSLKHHFHPV(1/24) NNPWHSLHSHTY(1/24) NPGHIGHRHHHT(1/24) WPYTRTHVHHVP(1/24) APHLKHHGLSFR(1/24) AWRHNHFTPVAQ(1/24) VGFHHLTEHPHR(1/24) RLQHQHFHPHVL(1/24) HLKIHHVRVDHM(1/24) TVRHHTHEVSNL(1/24) TYTHHSSSQHYG(1/24) LGSSHGHGASHQ(1/24) FHHH*TQRPAQG(1/24) 15 Phage display (subtractive panning) 5 PMID:30496775 Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Biopanning was performed as follows: 1. Immobilization of HER2 to Ni-NTA magnetic particles (MPs); 2. Blocking with BSA; 3. Phage binding; 4. Washing and elution of bound phage; 5. Amplification; 6. Negative selection (against BSA-blocked MPs). 3428 GQSEHHMRVASF(60/67)[0.42] GLHTSATNLYLH(3/67)[0.76] STPIFAEATARS(2/67)[1.20] SGVYKVAYDWQH(1/67)[0.33] ALKTHSVSPAPR(1/72)[1.11] 5 Phage display (subtractive panning) 5 PMID:30496775 Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance at 450 nm was measured after the reaction with the HRP-anti-M13 antibody conjugate (diluted 1:2000 in the blocking buffer). A450 was reproduced from Fig. 2 and shown. Biopanning was performed as follows: 1. Immobilization of HER2 to Ni-NTA magnetic particles (MPs); 2. Phage binding; 3. Washing and elution of bound phage; 4. Amplification; 5.Two negative selection steps (one against BSA-blocked MPs and another against Ni-NTA MPs). 3429 SGVYKVAYDWQH(37/57)[0.33] HLTTTHPEPPYG(11/57)[1.26] HRGDTTYNHLHP(4/57)[0.88] GQSEHHMRVASF(1/57)[0.42] YSHTLKIPAPDF(1/57)[1.43] 5 Phage display (subtractive panning) 5 PMID:30496775 Receptor tyrosine-protein kinase erbB-2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance at 450 nm was measured after the reaction with the HRP-anti-M13 antibody conjugate (diluted 1:2000 in the blocking buffer). A450 was reproduced from Fig. 2 and shown. Biopanning was performed as follows: 1. Binding between HER2 and phages; 2. Immobilization of HER2-phage binding mixture to Ni-NTA magnetic particles (MPs); 3. Washing and elution of bound phage; 4. Two negative selection steps (one against BSA-blocked MPs and another against Ni-NTA MPs). 3430 TPGGFAAVG TPGGFGLEL TPGGFLEIV TPGGFWGWM GPVGFSGLE GGSLFRGVV GSALFGIHH CSELFIQHH RVCLFMDMG EMSAFSSHH SYYAFSLVN PGGIFWFSL SVSFFMTHH VRWYFHHHH GTMYFLGGK GTMYFLGGK GTLMFLQRV TVSQFLSHH PGGSFLPLV GTVSFLTGN SVLSFMVGT DIIEFVGVH NEARFADQH GVVYFRGEA VPIDFRVPG GWESFKSLL EVSTFETEE YWALYRHHH VCELTTQEH QLGLYEFVV TPGGYLDYS VTLGYGVEH TPGGYSSMW SVLPYVTHH EDVPYSAHH VLEPYWSAQ GGEAYLSIG GGTVYWDVP RVGVYQEVS LCWQYSPHH YYTFYHHHH ESDFYSTHH SMMRYGEGM GPARYNSVL PGGMYSGTS HKADYSISA GAVVWGEGL SGSVWLDHH GAEVWMDAT GGLVWSYYI GRSVWSFMF HVAVWYAGH GGPLWAEGF GMNLWFAQD GWSLWFAGG GPVAWWYYP DIFAWHHHH PLAGWGHHH LSSFWNGEH RVISWMLPH PGGNWVSYS LVETWLTRP GTIEWAIRV 63 Phage display (common panning) 5 PMID:30458129 Cysteine protease CP16160 Not determined. Not determined. Not determined. X9 T7 phage display library Briefly, an aliquot of amplified phages (e9 pfu) were firstly bound to 125 μl of Ni–nitriloacetic acid (Ni–NTA) beads by their His6-tags for 1 h at 4 °C under gentle agitation. Unbound phages were removed by washing 10 times in 1.5 ml of 1M NaCl, 0.1% Tween-20 in PBS, pH 7.2 and two subsequent washes with 1.5 ml PBS. The Ni-NTA beads were then resuspended in 375 μl PBS. The purified CP (CP16160, ~800 ng) was added to the resuspended Ni-NTA beads to digest the susceptible phage nanopeptides under gentle agitation at 37 °C for 2 h. PBS without added enzyme was used as control. After 2 h incubation, phages with random peptide that was susceptible to protease cleavage were released from Ni-NTA matrix, and the supernatant containing these phages was recovered. To ensure that all of the released phages were recovered, the beads were re-suspended in 100 μl PBS (pH 7.2) and the supernatant, after mixing and centrifugation, was added to the first supernatant. To ensure that the His6-tags had been hydrolyzed on all phages recovered after protease digestion, 15 μl fresh Ni–NTA agarose beads were added to the combined phage supernatant and the mixture agitated for 15 min followed by centrifugation. A control elution of the phages still bound to the beads, using 100 μl 100 mM imidazole showed that at least 1e8 phages were attached to the matrix during each selection. 10 μl of the supernatant containing the released phages were used to determine the amount of phages detached in each round of selection. Dilutions of the supernatant were plated in 2.5 ml of 0.6% top agarose containing 300 μl of Escherichia coli (BLT5615), 100 μl diluted supernatant and 100 μl 100 mM isopropyl β-D-1-thiogalactopyranoside (IPTG). The remaining volume of the supernatant was added to a 10 ml culture of BLT5615 (OD ~0.6). The bacteria had 30 min prior to phage addition been induced to produce the T7 phage capsid protein by the addition of 100 μl 100 mM IPTG to the culture. The bacteria were lysed ~75 min after phage addition. The lysate was centrifuged to remove cell debris and 500 μl of the phage sub-library was added to 100 μl fresh Ni–NTA beads to start the next round of selection. After binding of the sub-library for 1 h at 4 °C under gentle agitation, the Ni–NTA beads were washed 15 times in 1.5 ml 1M NaCl, 0.1% Tween-20 in PBS, pH 7.2, followed by two subsequent washes with 1.5 ml PBS. 3431 ACPMNESKFC(12/22)[29.1] ACPSNPSKFC(6/22)[32.1] ACPKNPNKFC(2/22)[31.4] 3 Phage display (common panning) 3 PMID:30262856 Natalizumab Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Biolayer interferometry (BLI) To determine binding constants for association and dissociation of natalizumab with Ntz-01–07, biolayer interferometry (BLI) was performed by Antibody Solutions (Mountain View CA), using an Octet Red96 (Pall ForteBio LLC, Fremont CA). Briefly, 10 μg/ml of peptide was immobilized onto a streptavidin sensor. Three-fold dilutions of natalizumab were in solution, at concentrations from 1–600 nM (7 total dilutions). The assay steps were: sensor check (30 s), load antibody (600 s), baseline (300 s), antibody association (550 s), antibody dissociation (550 s). TBS was used as the buffer. KD (nM) for synthesized peptides was determined and shown. PKNPSKF is identified as a novel natalizumab-binding motif, and peptides containing this motif demonstrated utility as capture reagents in enzyme-linked immunosorbent assays (ELISAs). A peptide containing the identified motif was shown to be competitive with the natural ligand (α4-integrin) and a neutralizing anti-idiotype antibody for natalizumab binding. 3432 IYAAYPPCPQNLSKFCRHSSSPG(6/20)[NA] AYPHGRSCPQNISKFCFDHEKTN(5/20)[6.03] MPSPPKNCSKFHSALCKGVTWNV(2/20)[NA] SHPQEFWCPQNFSKFCSRSYSNT(1/20)[10.5] QGGEWHRCMSEEGKHCVDIQFIR(1/20)[NA] TSLTVMTCPHNPSKWCSPLPAAV(1/20)[NA] AMASSATCTKPNSYSCLHAKLVP(1/20)[NA] 7 Phage display (common panning) 3 PMID:30262856 Natalizumab Not determined. Not determined. Not determined. X7CX7CX7 M13 phage display library (X7CX7CX7) Biolayer interferometry (BLI) To determine binding constants for association and dissociation of natalizumab with Ntz-01–07, biolayer interferometry (BLI) was performed by Antibody Solutions (Mountain View CA), using an Octet Red96 (Pall ForteBio LLC, Fremont CA). Briefly, 10 μg/ml of peptide was immobilized onto a streptavidin sensor. Three-fold dilutions of natalizumab were in solution, at concentrations from 1–600 nM (7 total dilutions). The assay steps were: sensor check (30 s), load antibody (600 s), baseline (300 s), antibody association (550 s), antibody dissociation (550 s). TBS was used as the buffer. KD (nM) for synthesized peptides was determined and shown. 3433 QTLNHSWLHTFI(1/9) 1 Phage display (common panning) 3 PMID:30262856 Natalizumab Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3434 CSDRIMRGC[13.20] 1 Phage display (in vivo) 3 PMID:30082470 Human bladder cancer cell line RT112 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA For ELISA, RT112 and human normal urothelial cell line SV-HUC-1SV-HUC-1 were plated at 1.0e4 cells/well in a 96-well plate and incubated at 37°C in 5% CO2 for 24 hours. Subsequently, cells were washed and fixed with 4% paraformaldehyde for 15 minutes at room temperature. After incubation with 3% H2O2 (100 mL/well) at 37°C for 30 minutes, the plates were washed 3 times with PBS, and the wells were blocked with 200 mL of 5% BSA for 30 minutes at 37°C. The phages (1.0e10 PFU/well) were added to RT112 or SV-HUC-1 cells and incubated at 37°C for 2 hours. The plates were then washed 3 times with TBST before the addition of 100 mL of HRP-anti-M13 mAb (1:5,000) for 1 hour at 37°C. After washing 3 times with 0.05% TBST, 3,30,5,50-tetramethylbenzidine (TMB) was added at room temperature for 30 minutes. The reaction was terminated by the addition of 50 mL of 2 mol/L H2SO4. Subsequently, absorbance was read using an automated ELISA plate reader at 450 nm. Unrelated phages with equal titers were added to the wells in place of selected phage clones to serve as negative controls. Selectivity was determined using the following formula: Selectivity = [RT112(OD450 - OD450negative control)]/[SV-HUC-1(OD450 - OD450negative control)]. The selectivity value was reproduced from Figure 1A and shown. Subcutaneous tumor-bearing mice received 3e10 plaque-forming units (PFU) of the phage peptide library via tail-vein injection. After 15 minutes, mice were sacrificed and perfused by injection of 50 mL of PBS through the heart to wash unbound phages. Then, a portion of tumors was harvested and preserved in 4% paraformaldehyde and the rest was manually homogenized and washed with 10 × TBS containing 0.1% Tween-20 (0.1% TBST) to remove nonspecifically bound phages. Cell membrane–bound phages were eluted with 1 mL of elution buffer (0.2 mol/L glycine-HCl, pH 2.2, 1 mg/mL BSA) for 10 minutes on ice and neutralized with 150 mL of 1 mol/L Tris–HCl (pH 9.0). After centrifugation, the supernatant was collected, and the cells in the precipitate were washed once with PBS-BSA and lysed with 0.1% Triton X-100 for 2 hours at room temperature. Thus, the internalized phages in the cell lysate were recovered. A total of 10 mL of eluted phage was titrated on agar plates in the presence of IPTG/X-gel (1 mg/L) and tetracycline (40 mg/mL). The remaining phages were amplified by ER2738 bacteria at 37°C for 4.5 hours and injected into tumor-bearing mice. The biopanning process was repeated for three rounds. A bladder cancer–specific peptide with the sequence of CSDRIMRGC (PLSWT7) was selected by in vivo phage-display technology and labeled with IRDye800CW to synthesize a bladder cancer–specific dual-modality imaging (DMI) probe (PLSWT7-DMI). The feasibility of PLSWT7-DMI–based dual-modality photoacoustic imaging near-infrared (PAI-NIR) imaging was assessed in vitro, in mouse models, and ex vivo human bladders. An air-pouch bladder cancer (APBC) model suitable for probe instillation was established to evaluate the probe-based bladder cancer PAI diagnosis and NIR-imaging–guided resection. Human bladders were used to assess whether the PLSWT7-DMI–based DMI strategy is a translatable approach for bladder cancer detection and resection. The probe exhibited excellent selectivity and specificity both in vitro and in vivo. Postinstillation of the probe, tumors <3mm were detectable by PAI, and NIR-imaging–guided tumor resection decreased the bladder cancer recurrence rate by 90% and increased the survival in the mouse model. Additionally, ex vivo NIR imaging of human bladders indicated that PLSWT7-DMI–based imaging would potentially allow precise resection of bladder cancer in clinical settings. This PLSWT7-DMI–based DMI strategy was a translatable approach for bladder cancer diagnosis and resection and could potentially lower the bladder cancer recurrence rate. 3435 CNWMINKEC(3/20)[0.72 ± 0.09] CVPSKPGLC(3/20)[0.21 ± 0.02] CPKGDENTC(2/20)[0.12 ± 0.02] CPTSQRDNC(2/20)[0.16 ± 0.01] CVEKSAMSC(1/20)[NA] CLSTTEGYC(1/20)[NA] CIHSPTALC(1/20)[NA] CSLASTSHC(1/20)[NA] CLKPHSADC(1/20)[NA] CLHKSVSGC(1/20)[NA] CSSKHEATC(1/20)[NA] CSKMKIDHC(1/20)[NA] CSENSPLLC(1/20)[NA] CNQTEPFSC(1/20)[NA] 14 Phage display (subtractive panning) 3/4/5 PMID:30282451 Hepatitis A virus cellular receptor 1, HAVcr-1 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Individual phage clones were incubated with KIM-1‒coated plates and the bound phage clones were determined by ELISA. Plates were read at 450 nm using a microplate reader (Tecan, Zurich, Switzerland). The absorbance at 450 nm represent as the mean±SD of three separate experiments performed in triplicate, which was reproduced from Figrue 2A and shown. The absorbance of the wild type M13 phage was 0.10 ± 0.01. Each biopanning round consisted of subtraction of phages nonspecifically bound to BSA and subsequent selection of phages bound to KIM-1, followed by amplification of the eluted phages. A phage clone displaying the CNWMINKEC peptide showed higher binding affinity to KIM-1 and KIM-1-overexpressing 769-P renal tumor cells compared to other phage clones selected after biopanning. The CNWMINKEC peptide and a NeutrAvidin/biotin-CNWMINKEC multimer selectively bound to KIM-1 over albumin and to KIM-1-overexpressing 769-P cells and A549 lung tumor cells compared to KIM-1-low expressing HEK293 normal cells. Co-localization and competition assays using an anti-KIM-1 antibody demonstrated that the binding of the CNWMINKEC peptide to 769-P cells was specifically mediated by KIM-1. The CNWMINKEC peptide was not cytotoxic to cells and was stable for up to 24 hours in the presence of serum. Whole-body fluorescence imaging demonstrated selective homing of the CNWMINKEC peptide to KIM-1‒overexpressing A498 renal tumor compared to KIM-1‒low expressing HepG2 liver tumor in mice. 3436 YHMEFWDELWGI(4)[0.55 ± 0.11] WHNPLWWLSAYE(3)[0.61 ± 0.09] WHEYPLVWLSGY(3)[0.47 ± 0.14] 3 Phage display (common panning) 4 PMID:30269789 Glycoprotein 4, Protein GP4 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities between the selected phages and GP4 recombinant protein were determined by phage ELISA. OD405 was detected with an ELISA plate reader. The original mixed phage library was used as the negative control. The OD405 value was reproduced from Figure 2A and shown. Phage display biopanning on GP4 protein showed that the specific phages obtained could distinguish porcine reproductive and respiratory syndrome virus (PRRSV) from the other viruses. The exogenous peptide WHEYPLVWLSGY displayed on one of the candidate phages showed high affinity for GP4 protein and exerted a significant inhibitory effect on PRRSV penetration in vitro. Moreover, the N-terminus of GP4 was predicted as the critical receptor binding site and the beginning of the fifth scavenger receptor cysteine-rich domain of CD163 as the critical ligand recognition site based on sequence alignment and model prediction analyses. 3437 GLHTSATNLYLH(13)[12.6] SGVYKVAYDWQH(6)[5.7] FIPFDPMSMRWE(3)[7.2] ALAVAPSRWWNE(3)[76.3] KASGSPSGFWPS(3)[NA] NASSFPTNSRWA(2)[20.6] QFDYMRPANDTH(2)[3.3] GQSEHHMRVASF(2)[NA] GTGLVTLPRLTV(2)[NA] DLILLNGGSQQA(1)[NA] LPVGNQSDVHFD(1)[NA] SPLCCNSGTDQT(1)[NA] DPIKHLEDQAST(1)[NA] AAIEPSAGLAYF(1)[NA] SPYTWGSGDWRS(1)[NA] DQFVHDVKGTKH(1)[NA] AHWGMSARTSAW(1)[NA] TNRTHNINALHF(1)[NA] GNHNNSVTTWHL(1)[NA] MHPNAGHGSLMR(1)[NA] TPGGLLRNDRPL(1)[NA] NIYTTPWGSNWS(1)[NA] HLARTTDMLHPY(1)[NA] QMSPGSTARSAL(1)[NA] TVQSLPPHSPYD(1)[NA] MDNKTTRWRDPL(1)[NA] HSIALTTTLRTP(1)[NA] MSKSTMSLSSNA(1)[NA] TSIQISNAHPKS(1)[NA] HTSYGTTWSESR(1)[NA] NHLSTPVWSITG(1)[NA] WSMNHTNLSYTS(1)[NA] QPDFTPRPAYSR(1)[NA] ITGCDPTKQTQG(1)[NA] AYPQKFNNNFMS(1)[NA] YDIHWSPDSRLK(1)[NA] ASATVSKPWPTG(1)[NA] KVMGLSVTSHAE(1)[NA] SNSSLELPNLNS(1)[NA] YIWQGPGSAGTP(1)[NA] HGNPRSPNQFST(1)[NA] QCFNSVCLHTNP(1)[NA] QQHPLITRLISH(1)[NA] ALEVNGMRPAWA(1)[NA] HRGDTTYNHLHP(1)[NA] SQGLSLSTMVTR(1)[NA] ADPLGISYLRSS(1)[NA] KATDFPDGRLNY(1)[NA] SAELPHKTGSVL(1)[NA] GSAPLLTVDTSK(1)[NA] STPIFAEATARS(1)[NA] LPHLPSQNVRAP(1)[NA] SCDMQERCLAGL(1)[NA] VLYEQVLAAPME(1)[NA] TAGAIADFQSLT(1)[NA] LTHYPVTGSTGS(1)[NA] SEFQSTHWLDSL(1)[NA] WIRPPSGPMYSF(1)[NA] DPTRQEPHLFLL(1)[NA] QAMMWLPVALAA(1)[NA] SGKLLMLPVVSR(1)[NA] YTTIRWSEFSAW(1)[NA] DRWVARDPASIF(1)[NA] ISSTDRLLAAGL(1)[NA] SGTSLLHYHSYH(1)[NA] DSQFNKYSIATV(1)[NA] SNLSARNTISHG(1)[NA] MDGSSREVNDAV(1)[NA] 68 Phage display (common panning) 4 PMID:29956504 Human breast cancer cell line MCF7 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Flow Cytometry The fluorescence intensity of each FITC-labeled peptide at 1.0e–6 m (also defined as the fluorescence per unit, FPU) was measured by a spectrofluorometer (VARIAN, Cary Eclipse, USA, Ex = 488 nm, Em = 535 nm). The FPU was used for calibration of the relative affinity. The mean fluorescence intensity (MFI) of 10000 cells was measured by a FACSCalibur flow cytometer (Becton Dickinson, USA). MCF7 cells incubated with blank RPMI-1640 medium were taken as the background group and MCF7 cells incubated with FITC-AGP were used as the control group. The relative affinity of a peptide to MCF7 cells was calculated using the following formula: Relative affinity = ((MFIpeptide -MFIbackground)/FPUpeptide)/((MFIcontrol - MFIbackground)/FPUcontrol), which means the affinity ratio of the given peptide to the control peptide. The mean relative affinity of ligand peptides to MCF7 cells was shown in Figure 2A and shown. MCF7 cells were seeded in a 25 cm2 flask and cultured until confluence (about 3 .0e6 cells per flask). Then the cells were rinsed twice with PBS and blocked with 0.5% bovine serum albumin in RPMI-1640 medium (BSA-1640) at 4 °C for 30 min. After blocking, the cells were coincubated with phages (1.0e11 pfu, Ph.D.-12 peptide library kit, the input phage library, displaying a diversity of 1.0e9 dodecapeptides) in BSA-1640 at 4 °C for 30 min to allow for binding or internalization between phages and cells. The medium containing unbound phages was discarded and the cells were washed ten times with BSA-1640 containing 0.1% Tween-20 to remove weakly bound phages. Afterward, the phages that bound strongly to MCF7 cells were eluted down with Glycine-HCl (2 m, pH 2.2) solution and the acidic eluate was recovered as one output phage sublibrary, which had higher cell-binding affinity to MCF7 cells than the input phage library. Finally, after washing thrice, the cells were lysed by Tris-EDTA solution (50e−3 m Tris, 1.0e−3 m EDTA, 150e−3 m NaCl, 2% deoxycholate, pH 10.0) and the cell lysate containing intracellular phages was recovered as another output phage sublibrary, which had higher cell-internalizing efficacy to MCF7 cells than the input phage library. The phage titer was assayed by counting plague forming units. The recovery rate of phage in each round was calculated as the ratio of output to input. The two output phage sublibraries (eluate and lysate) were amplified and used as the input phage libraries of the next panning round, respectively. A total of four rounds were performed to achieve phage sublibraries with sufficient affinity to MCF7 cells. Four of these ligand peptides [FIPFDPMSMRWE (FIP), NASSFPTNSRWA (NAS), GLHTSATNLYLH (GLH), and ALAVAPSRWWNE (ALA), respectively] exhibit high affinity to MCF7 human breast cancer cells. Among them, NAS and ALA are reported for the first time, whose affinities are 20.6 and 76.3 times that of the random peptide control, respectively. Both NAS and ALA modifications to doxorubicin-loaded lipid nanosytems [LP(DOX)] show stronger tumor inhibition, longer animal survival time, and less body weight loss, compared to unmodified or control peptide modified nanosystems, on an MCF7 tumor-bearing mouse model. 3438 CYLLCISPC CGSRCYPRC 2 Phage display (subtractive panning) 1 PMID:30198459 IgG from tegumentary leishmaniasis (TL) patients Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Briefly, viral particles of the bacteriophage library (Ph.D.®-C7C library, New England BioLabs, Ipswich, MA, USA) were diluted in 250 μL of 50mM Tris-HCl pH 7.5, 150 mM NaCl and 0.1% Tween 20 buffer (TBS-T). The mixture was incubated for 30 min, at room temperature, with microspheres coupled to the IgG antibodies purified from healthy subjects for subtraction, which were magnetically captured. The remaining phages in the supernatant were recovered and transferred to a new tube, which was subjected to positive selection using IgG from TL patients. The supernatant was removed and the bound phages were washed five times in 1 mL TBS-T buffer, and were eluted in 500 μL of 0.2 M glycine buffer, pH 2.0. Next, 75 μL of 1 M Tris-base pH 9.0 were added to neutralize the acidic pH of the solution. Two clones, namely A4 (CYLLCISPC) and A8 (CGSRCYPRC), were identified and used in immunization protocols from BALB/c mice to protect against Leishmania amazonensis infection. Results showed a polarized Th1 response generated after vaccination, being based on significantly higher levels of interferon-γ (IFN-γ), interleukin-2 (IL-2), IL-12, tumour necrosis factor-α (TNF-α) and granulocyte-macrophage colony-stimulating factor (GM-CSF); which were associated with lower production of specific IL-4, IL-10 and immunoglobulin G1 (IgG1) antibodies. Vaccinated mice presented significant reductions in the parasite load in the infected tissue and distinct organs, when compared with controls. 3439 GWIVSFAVQTEGHGVAD(70) GGTQCLQFKVIPAGHFC(40) GIARSNQDIFVYDAHKC(9) GYYPSVGHQTCQDIFVA(8) GYIVSQTEGRHPQHQEA(4) GAGFCYQQYLEQDIFVY(2) GWKNCAVVQTEGCNCYF(2) GSIWSQVQTEGYRHNHA(2) GEELCQTEGRGPEVQPY(2) GNLACWYNEQDIFVAHD(1) GVFGSITVVQTEGVYSC(1) GTWSCRYYQQVQTEGQD(1) GVNVSVQTEGAWHAQAY(1) GVHMCKLHLLVQTEGQC(1) GQPTCYNHNKQQTEGRC(1) 15 Phage display (competitive panning) 2 PMID:30267550 Serum 13003 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 3440 GYQWSTEHIFVIPCQRA(4) GEPQSEQDIFQKCEKQF(1) GTFHSIQQQRDUFVIVI(1) 3 Phage display (competitive panning) 2 PMID:30267550 Serum 14008 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 3441 GSENCDIFVIQEIQSCF(3) GLNRCDIFVIPECELHN(2) 2 Phage display (competitive panning) 2 PMID:30267550 Serum 13009 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 3442 GSWPSLEIFVIPIFVQI(112) 1 Phage display (competitive panning) 2 PMID:30267550 Serum 13007 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 3443 GIRACWQAPFVIPAGIC(1) 1 Phage display (competitive panning) 2 PMID:30267550 Serum 13016 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 3444 GSAVCSYGFVIPAGCII(5) GIWGSPAGYPEECVVRD(3) 2 Phage display (competitive panning) 2 PMID:30267550 Serum 14012 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 3445 GLVSSQLTQFVIPAHCD(2) 1 Phage display (competitive panning) 2 PMID:30267550 Serum 13002 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 3446 GTSQSEAVIPAGHHEHY(15) GEKHSEDPFLVIPAGGD(8) 2 Phage display (competitive panning) 2 PMID:30267550 Serum 13021 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 3447 GIQRSAGYPVFKDVDID(4) 1 Phage display (competitive panning) 2 PMID:30267550 Serum 13001 from patients sensitized to soy bean Not determined. Not determined. Not determined. ENTE-1 phage display library (GX3[C/S]X12) A trinucleotide‐based, 16mer peptide phagemid library covering >e9 clones (“ENTE‐1”) was used for two rounds of panning against individual serum from patients sensitized to soy bean. We coated 24 μg total serum protein in 2.5 mL PBS to Immuno Tubes (MaxiSorp™; Thermo Fisher Scientific, Waltham, MA, USA) for 1 hour at 4°C under constant agitation. Under these conditions, all Ig molecules should be bound on the tube surface. A negative control was incubated with PBS only. The coated tubes were washed with 2.5 mL blocking buffer once (5% [w/v] non‐fat dry milk powder in PBS) and incubated with 4 mL blocking buffer at 4°C for at least 15 minutes under constant agitation. Before adding the ENTE‐1 peptide phage display library, the tubes were washed three times with 4 mL PBS. To preferably select patient-specific antibodies, and in particular to prevent selection of peptides binding serum proteins, a competition with a mixture of ten sera from healthy donors was conducted. The ENTE‐1 library was added to each tube in 1 mL blocking buffer containing 30 μg of non‐allergic serum protein. In the first selection round, an amount of phage corresponding to 4e11 colony forming units (cfu) of ENTE‐1 library and in the second selection only e9‐e10 cfu of recovered phage were used. Tubes were incubated for 2 hour at 4°C with constant agitation, washed five times with 1 mL 0.1% (v/ v) Tween 20 in PBS in the first selection round, and with 0.5% (v/v) Tween 20 in PBS in the second selection round. 3448 GTMPDKDTGMRQ ERAGTMPDRPRH GVMLSDRLERAL FQARWEPPRLLQ HSNSYQASVLCQ TLMQPSHAGLTL GGHQVNQLPSQM YSPRLSTLQSTP SSSGNPLYSAKI TLSLPGFTFVPT HTVEPRWVPRSW FLVFLVCTVHPP AVCTFCASDWWP NIWTPSLSLTYD FGDPFVSALFPQ SAPVPGSSHSRQ KPSVPGCTLVPS NCSNSIPCAVKA QAILYSPDRNPL TITNAPIKDLTP QVNGLGERSQQM TLQPQSRDLSTL YLTPELFLRTQQ MLQDNLQNLDFR LNPNWTYRSTNE HEASRLLGARLT VSKAPSKLETY 27 Phage display (common panning) 3 PMID:30187588 IgE from patients with allergy to Scylla paramamosain AK Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Overnight, coating of the wells of microtiter plate (Nunc Maxisorp, Copenhagen, Denmark) at 4°C with goat anti-human IgE (ε‐chain specific) (Sigma‐Aldrich, St. Louis, MO, USA) was used to capture IgE from patients’ sera. The phage display peptide library was added after the blocking incubation with captured IgE and eluted after the washing steps. 3449 VGSYSATVHRIQ VLAHKYTTGSLP ACLYCYDLVDSV SLPLTVVPTANR TLGLRPVPVATT TNSFHGIAGYQS SSLLGYNLNANH ASHTGTAQISRQ ETLKPYPLPNNS DGMLSLSLPASF VRATQYPMLLDF NGPSDLKYLHPT HPHDRQLISTYP NDVGVLSPETRQ IEQRHQTHSLHP HMSYGNGTRDTP DNHAKLPLRGSS VVFDGTLYNYHW DIARTAKPSHWP AYTVDALHELRH SYEPNSTKQFAP VDDLVRVSVNFR MYNPDPNRLARS WHYNWQDVSDRQ NMGGLDYLNKNS EALTVNIKREME HLTATELANSYH YTLSDRSHNPFN SWIPWAAFEHKQ WTTAAPAKSEYH TPVQQGQTKNAF FMIPGSMPYIGC CLYCWQGTSQQR WDWPLFPLDTLR AYEPSSHKSEVP VPIHAAHGFWHY WPAPFHNLVNPR 37 Phage display (common panning) 3 PMID:30187588 IgE from patients with allergy to Scylla paramamosain SCP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Overnight, coating of the wells of microtiter plate (Nunc Maxisorp, Copenhagen, Denmark) at 4°C with goat anti-human IgE (ε‐chain specific) (Sigma‐Aldrich, St. Louis, MO, USA) was used to capture IgE from patients’ sera. The phage display peptide library was added after the blocking incubation with captured IgE and eluted after the washing steps. 3450 SVDGWLEPPTST DIARTAKPSHWP DINIRVYPSQFP LVPSPQSAMRFD AHDSADYLNAPR SHNAPWSPTLAL YPGISHVGSKVS TGSAKFLQRDTH TLMQPSHAGLTL DQYLMTTTRGAP YPGSQSWMPSDF GCNHDSCSALTK DNHAKLPLRGSS TIPTRDPAMLHS 7 Phage display (common panning) 3 PMID:30187588 IgE from patients with allergy to Scylla paramamosain TIM Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Overnight, coating of the wells of microtiter plate (Nunc Maxisorp, Copenhagen, Denmark) at 4°C with goat anti-human IgE (ε‐chain specific) (Sigma‐Aldrich, St. Louis, MO, USA) was used to capture IgE from patients’ sera. The phage display peptide library was added after the blocking incubation with captured IgE and eluted after the washing steps. 3451 SVEPSDSSNPIY HFGVDPNADFVS FRVDLKNDGMPE SPALHTTIPGAK YHAKNDGMVFPT TYNYDMPLRGRA TIPTRDPAMLHS TALPKVHTLLTH NPHDDVLWLHPS VPLDHAYGHSIP AYTVDALHELRH VGPLGQVMSGHS HPHDRQLISTYP 13 Phage display (common panning) 3 PMID:30187588 IgE from patients with allergy to Scylla paramamosain FLN c Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Overnight, coating of the wells of microtiter plate (Nunc Maxisorp, Copenhagen, Denmark) at 4°C with goat anti-human IgE (ε‐chain specific) (Sigma‐Aldrich, St. Louis, MO, USA) was used to capture IgE from patients’ sera. The phage display peptide library was added after the blocking incubation with captured IgE and eluted after the washing steps. 3452 CLHQSPHIC(4/44) CPTNNPRSC(2/44) CAQIPRANC(2/44) CKLTHPQTC(2/44) CMNTDKRMC(2/44) CQPANSKMC(2/44) CDTRWSNLC(1/44) CEQRTSHKC(1/44) CFGSTTWKC(1/44) CGLAKETMC(1/44) CHTHPTHDC(1/44) CKSHAHQNC(1/44) CLHHNNAYC(1/44) CMGMGQAWC(1/44) CMTKMAPHC(1/44) CMHNGSWQC(1/44) CNKVHGKTC(1/44) CNSHQASAC(1/44) CNTKGPYQC(1/44) CPKPHSDTC(1/44) CPQNGKEVC(1/44) CPSNKLTQC(1/44) CRYGLEHKC(1/44) CRVNDQSQC(1/44) CSRSHHDHC(1/44) CSPALIGQC(1/44) CSKHRIHQC(1/44) CSQSYRHSC(1/44) CSSHPALRC(1/44) CSNTSLNAC(1/44) CSHSSMQKC(1/44) CTGKSAGSC(1/44) CTSANLRNC(1/44) CTSPSRRAC(1/44) CTTAKHTTC(1/44) CYTSPKGRC(1/44) 36 Phage display (in vivo) 3 PMID:31193742 Spinal Cord Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The CX7C phage library was used for in vivo phage display for the screening of spinal cord homing peptides. The phage library was injected into C57BL/6 mice through the tail vein. At 5 min after injections, whole spinal cords were isolated after transcardiac removal of blood. Aliquots from 3–5 mice were combined and infected into Escherichia coli ER2738. The number of phages included in the spinal cord was titrated as PFU. After amplification to 1.0e11 PFU, the phages were again injected into other C57BL/6 mice. This biopanning procedure was repeated three times to select phages that specifically bound to the spinal cord. After the third round of biopanning, each phage genome was isolated from phage plaques and analyzed using DNA sequence-coding pIII protein. Analyses of the sequences of targeted phages yielded two candidate peptides targeting the spinal cord: SP1 (CLHQSPHIC) and SP2 (CPTNNPRSC). These peptides were synthesized and intravenously injected into mice to evaluate their tissue specificity and potential as gene delivery carriers. The complexes between SP1 or SP2 peptides and the plasmid vector expressing the reporter gene could induce gene transduction in the spinal cord through systemic injection without gene expression in the brain, liver, and kidney. In addition, intravenous injection of the complex between SP1 and the vectors induced interleukin-4 expression in the spinal cord, resulting in effective suppression of lipopolysaccharide-induced hyperalgesia. Therefore, intravenously administered spinal cord homing peptides complexed with a plasmid vector provided tissue-specific treatment featuring gene delivery to the CNS through systemic circulation. 3453 CLHQSPHIC(8/49) CPTNNPRSC(6/49) CFGQKASSC(2/49) CLTSTHMWC(2/49) CSKHGMPYC(2/49) CTSPSRRAC(2/49) CDLPWIQSC(1/49) CFGSTTWKC(1/49) CGLAKETMC(1/49) CGPHQFNLC(1/49) CHGPGHHSC(1/49) CHNPSIHNC(1/49) CKTYPSHLC(1/49) CKQTQTHFC(1/49) CLNPKTQYC(1/49) CMGMGQAWC(1/49) CMHNGSWQC(1/49) CMMETPGSC(1/49) CMMDHTLQC(1/49) CMTKMAPHC(1/49) CNHHSGLTC(1/49) CNLSSNRQC(1/49) CNNMPAKVC(1/49) CNPMGKLQC(1/49) CNTRAPSTC(1/49) CPGLCAHKC(1/49) CPTATPQLC(1/49) CRSANIYTC(1/49) CSQNQPKVC(1/49) CTHSSLPIC(1/49) CTSKPAHYC(1/49) CTYRPPHLC(1/49) CVPSRLPLC(1/49) 33 Phage display (in vivo) 4 PMID:31193742 Spinal Cord Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The CX7C phage library was used for in vivo phage display for the screening of spinal cord homing peptides. The phage library was injected into C57BL/6 mice through the tail vein. At 5 min after injections, whole spinal cords were isolated after transcardiac removal of blood. Aliquots from 3–5 mice were combined and infected into Escherichia coli ER2738. The number of phages included in the spinal cord was titrated as PFU. After amplification to 1.0e11 PFU, the phages were again injected into other C57BL/6 mice. This biopanning procedure was repeated four times to select phages that specifically bound to the spinal cord. After the fourth round of biopanning, each phage genome was isolated from phage plaques and analyzed using DNA sequence-coding pIII protein. Analyses of the sequences of targeted phages yielded two candidate peptides targeting the spinal cord: SP1 (CLHQSPHIC) and SP2 (CPTNNPRSC). These peptides were synthesized and intravenously injected into mice to evaluate their tissue specificity and potential as gene delivery carriers. The complexes between SP1 or SP2 peptides and the plasmid vector expressing the reporter gene could induce gene transduction in the spinal cord through systemic injection without gene expression in the brain, liver, and kidney. In addition, intravenous injection of the complex between SP1 and the vectors induced interleukin-4 expression in the spinal cord, resulting in effective suppression of lipopolysaccharide-induced hyperalgesia. Therefore, intravenously administered spinal cord homing peptides complexed with a plasmid vector provided tissue-specific treatment featuring gene delivery to the CNS through systemic circulation. 3454 CLHQSPHIC(31/47) CPTNNPRSC(12/47) CNMRTLMQC(2/47) CDMHQGKTC(1/47) CSPDKNRSC(1/47) 5 Phage display (in vivo) 5 PMID:31193742 Spinal Cord Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The CX7C phage library was used for in vivo phage display for the screening of spinal cord homing peptides. The phage library was injected into C57BL/6 mice through the tail vein. At 5 min after injections, whole spinal cords were isolated after transcardiac removal of blood. Aliquots from 3–5 mice were combined and infected into Escherichia coli ER2738. The number of phages included in the spinal cord was titrated as PFU. After amplification to 1.0e11 PFU, the phages were again injected into other C57BL/6 mice. This biopanning procedure was repeated five times to select phages that specifically bound to the spinal cord. After the fifth round of biopanning, each phage genome was isolated from phage plaques and analyzed using DNA sequence-coding pIII protein. Analyses of the sequences of targeted phages yielded two candidate peptides targeting the spinal cord: SP1 (CLHQSPHIC) and SP2 (CPTNNPRSC). These peptides were synthesized and intravenously injected into mice to evaluate their tissue specificity and potential as gene delivery carriers. The complexes between SP1 or SP2 peptides and the plasmid vector expressing the reporter gene could induce gene transduction in the spinal cord through systemic injection without gene expression in the brain, liver, and kidney. In addition, intravenous injection of the complex between SP1 and the vectors induced interleukin-4 expression in the spinal cord, resulting in effective suppression of lipopolysaccharide-induced hyperalgesia. Therefore, intravenously administered spinal cord homing peptides complexed with a plasmid vector provided tissue-specific treatment featuring gene delivery to the CNS through systemic circulation. 3455 CTTNPFSLC(5/12)[2913.0±311.0] CRLSMETVC(3/12)[NA] CQAPHKPWC(1/12)[NA] CHNSKSTTC(1/12)[NA] CPSSMRGTC(1/12)[NA] CILKKNVSC(1/12)[NA] 6 Phage display (common panning) 2 PMID:31173215 Bone mesenchymal stem cell (BMSC) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Flow Cytometry The bone mesenchymal stem cell (BMSC) affinity properties of FITC-labeled peptides were analyzed quantitively by flow cytometry at a wavelength of 488 nm, using FlowJo v10 (Tree Star, Inc). A specific cyclic peptide for Sprague-Dawley rat bone mesenchymal stem cells (BMSCs), CTTNPFSLC (known as C7), was used, which was identified via phage display technology. Its high affinity for BMSCs was demonstrated using flow cytometry and fluorescence staining. Subsequently, the cyclic peptide was placed on β-tricalcium phosphate (β-TCP) scaffolds using absorption and freeze-drying processes. Adhesion, expansion and proliferation of BMSCs was investigated in vitro on the C7-treated β-TCP scaffolds and compared with pure β-TCP scaffolds. The results revealed that C7 had a promoting effect on the adhesion, expansion and proliferation of BMSCs on β-TCP scaffolds. Therefore, C7 may be effective in future tissue engineering therapy for osteonecrosis of the femoral head (ONFH). 3456 CTTNPFSLC(6/12)[2913.0 ± 311.0] CQAPHKPWC(2/12)[NA] CRLSMETVC(1/12)[NA] CPSSMRGTC(1/12)[NA] CDLLESERC(1/12)[NA] CKMWNGSGC(1/12)[NA] 6 Phage display (common panning) 3 PMID:31173215 Bone mesenchymal stem cell (BMSC) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Flow Cytometry The bone mesenchymal stem cell (BMSC) affinity properties of FITC-labeled peptides were analyzed quantitively by flow cytometry at a wavelength of 488 nm, using FlowJo v10 (Tree Star, Inc). A specific cyclic peptide for Sprague-Dawley rat bone mesenchymal stem cells (BMSCs), CTTNPFSLC (known as C7), was used, which was identified via phage display technology. Its high affinity for BMSCs was demonstrated using flow cytometry and fluorescence staining. Subsequently, the cyclic peptide was placed on β-tricalcium phosphate (β-TCP) scaffolds using absorption and freeze-drying processes. Adhesion, expansion and proliferation of BMSCs was investigated in vitro on the C7-treated β-TCP scaffolds and compared with pure β-TCP scaffolds. The results revealed that C7 had a promoting effect on the adhesion, expansion and proliferation of BMSCs on β-TCP scaffolds. Therefore, C7 may be effective in future tissue engineering therapy for osteonecrosis of the femoral head (ONFH). 3457 CVPSKPGLC[60] 1 Phage display (competitive panning) 3 PMID:31152222 Anti-aflatoxin B1 (AFB1) monoclonal antibody (MAb) 2F5 Aflatoxin B1 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA IC50 (ng/mL) was calculated in competitive phage ELISA. The competitive panning method was employed in this study. In brief, one well of a microtiter plate was coated with 100 μL of MAb 2F5 and incubated overnight at 4 °C. The next day, the phage library diluted with PBS was first added to the MAb 2F5-coated well and incubated for 1 h at 25 °C. After washing with PBST, the combined peptide was competitively eluted using 500 ng/mL AFB1 in 10% methanol PBS. Subsequently, the supernatants of the four wells were collected and infected into E. coli ER2738 for amplification by mixing with PEG solution. The entire panning procedure was repeated twice, except for the use of concentrations of AFB1, MAb 2F5, and Tween 20 in PBST in the second and third round of panning, respectively. After each round of panning, the eluted phage titer was determined according to the manufacturer’s protocol. The phage DNA was sequenced by Sangon Biotech using 96 gIII primer. 3458 LPSYPGYYQITI(7)[1.860 ± 0.056] CWTISLFGGVTQ(3)[1.576 ± 0.055] SKRSTGKDYTAT(1)[1.461 ± 0.078] RRVISARWTSDR(1)[1.461 ± 0.055] MQTSFLKHHSVT(1)[1.306 ± 0.066] 5 Phage display (common panning) 4 PMID:31149727 Apical membrane antigen-1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The optical density (OD) was measured at 490 nm using a microplate reader. The OD values were reproduced from Figure 2 and shown. Two corresponding specific Eimeria tenella apical membrane antigen-1 (EtAMA1) binding peptides (LPSYPGYYQITI (L) and CWTISLFGGVTQ (C)) showed significant effects on inhibiting sporozoite invasion of MDBK cells. 3459 ITMSVPAHNAKE(39/169)[34.2 ± 2.11] TNTSWDPQYNPD(11/169)[80.1 ± 1.13] NHFVPTSNRFNA(11/169)[120 ± 1.23] NFTINGKTHRLW(6/169)[298 ± 1.45] NAITLLSPPLHK(5/169)[57.1 ± 1.77] SSHNHDSYHGTK(2/169)[476 ± 2.73] LMNPATMKTSSG(1/169)[360 ± 1.87] TNTSWDPQYNPD(1/169)[47.6 ± 2.51] SNMKPSMEYSSR(1/169)[273 ± 1.79] IGNSWPLTSHSW(1/169)[144 ± 1.1] SYNTFMYERASK(1/169)[562 ± 1.69] MVHSKASMWPGK(1/169)[690 ± 2.27] KVYAINSWTNYY(1/169)[1200 ± 90.2] 13 Phage display (competitive panning) 5 PMID:31146360 Tubulin alpha chain Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding affinities of phages were determined by the enzyme-linked immunosorbent assay (ELISA) method with anti-M13 phage antibody. The optical densities were measured at 450 nm with a Biotrak multiwall plate reader (Amersham Bioscience). Kd (pM) was calculated and shown. In the final round, bound phages were eluted by addition of a 10 M excess of tubulin alpha chain. One peptide with the sequence of ITMSVPAHNAKEK demonstrated the high inhibitory effect on microtubule formation with a nanomolar range of IC50 values, which were much lower than a well-known chemical inhibitor—benomyl. Based on these results, this peptide can be employed to further develop promising candidates for novel antifungal agents against Phytophthora blight. 3460 TNPQARWHEYNF(61/112)[41.9 ± 1.05] NPIGDNYSGTGL(10/112)[37.1 ± 1.41] 2 Phage display (competitive panning) 5 PMID:31146360 Tubulin beta chain Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding affinities of phages were determined by the enzyme-linked immunosorbent assay (ELISA) method with anti-M13 phage antibody. The optical densities were measured at 450 nm with a Biotrak multiwall plate reader (Amersham Bioscience). Kd (pM) was calculated and shown. In the final round, bound phages were eluted by addition of a 10 M excess of tubulin beta chain. 3461 KALLHHLALLALHLA[0.46] WIALHHLLHLAAHWI[4.2] WALAHKALHALAHKP[5.3] KWLAKHAAGLALHAL[4.6] VLALHHALALAHKKA[9.0] PAALHHALALAHHLW[12.0] WMHKHQALAAMHAHR[0.12] RQAHTHALHHLALWC[0.13] WRLHHRHFLALALKR[0.21] TPHLHMFHAHKLAPR[0.16] TPHAHMWHAHKRNPK[0.56] PVHHVHLTAHHAVGC[1.5] 12 Phage display (common panning) 5 PMID:12849992 Lipid A Not determined. Not determined. Not determined. f5-15mer phage display library (X15) Surface plasmon resonance (SPR) Hybrid bilayers containing 20 mol% lipid A in DMPC on HPA chip were formed. The rate constants were determined by the nonlinear least squares fitting of the primary sensogram data using BiaEvaluation, Version 3.0 software. The association constant Ka (Unit × e6 1/M) was determined and shown. The lipid A solution was then sonicated for 5 min with a needle probe. This solution (2.0 ml) was then added to polystyrene petri dishes and incubated at 60 ℃. Subsequently, the dishes were washed five times with TBS (50 mM Tris–HCl, pH 7.5, and 150 mM NaCl) and blocked with 5 mg/ml of BSA for 2 h. Phages (2.0e11) were pipetted into each coated and blocked dish and were kept for 2 h at 4 ℃. Unbound phages were washed off with TBS and the bound phages were eluted with 0.1 M glycine HCl buffer (pH 2.2) containing 1 mg/ml BSA and neutralized with 0.1 M Tris–HCl, pH 9.0. The eluate was titered and amplified for use in subsequent round of panning, which were carried out in similarly coated wells of a 96-well microtitre plate. A comparison of the sequences revealed no consensus sequence between the 12 selected peptides suggesting that the lipid A binding motif is not sequence specific which is in accord with the sequence variation seen with the naturally occurring anti-microbial and/or endotoxin binding peptides. 3462 RLCSWISPCSA(4)[6] FGCSWLFPCPF(2)[8] RLCSWVSPCSA(1)[10] 3 Phage display (common panning) 3 PMID:15233528 B-domain deleted form of human factor VIII (BDDrFVIII) Not determined. Not determined. Not determined. TN7/1 phage display library ELISA Upon completion of the second and third rounds of selection, randomly-chosen individual phage isolates from pools generated by both elution conditions were screened for binding to BDDrFVIII by indirect phage ELISA. Background binding levels were established using wells coated with streptavidin alone or with BSA. Phage isolates exhibiting ELISA signals significantly above background (A630 > 0.25) were scored as positive for binding to BDDrFVIII. The ratio of the ELISA signal with target to the ELISA signal without target was shown. Biotinylated BDDrFVIII protein that was confirmed to be active after derivatization was immobilized onto streptavidin-coated magnetic beads and used as a target in six separate selection campaigns. Each campaign incorporated three rounds of selection from a single library, using one elution condition for phage bound to the immobilized BDDrFVIII. A round of selection consisted of a binding step in which the phage library was first incubated in suspension with the target, followed by a plate wash step to remove unbound or weakly-bound members of the phage library, and an elution step to recover a pool of phage. This results in a population of phage isolates enriched in members that bind the target under binding and wash conditions and release the target under elution conditions. Finally, elution pools from each round of selection were separately amplified by infection and growth in E. coli cultures. The purified phage preparations from this amplification step served as the input for subsequent rounds of selection. The consensus sequence is rlCSWѲsPCsa (uppercase letters stronger than lowercase, Ѳ = V, I, L, F). 3463 HPCGSWLRPCLH(10)[16] HSCGSWLFPCFA(7)[10] HLCGAWFRPCDA(6)[6] HPCGSWFNPCAH(4)[8] HPCGSWFRPCFH(3)[16] HACGSWFRPCHA(3)[6] HPCGAWLRPCYN(1)[20] HLCFAWFRPCDA(1)[8] HPCGSWLHPCAA(1)[6] HRCGSWLHPCLA(1)[6] 10 Phage display (common panning) 3 PMID:15233528 B-domain deleted form of human factor VIII (BDDrFVIII) Not determined. Not determined. Not determined. TN8/6 phage display library ELISA Upon completion of the second and third rounds of selection, randomly-chosen individual phage isolates from pools generated by both elution conditions were screened for binding to BDDrFVIII by indirect phage ELISA. Background binding levels were established using wells coated with streptavidin alone or with BSA. Phage isolates exhibiting ELISA signals significantly above background (A630 > 0.25) were scored as positive for binding to BDDrFVIII. The ratio of the ELISA signal with target to the ELISA signal without target was shown. Biotinylated BDDrFVIII protein that was confirmed to be active after derivatization was immobilized onto streptavidin-coated magnetic beads and used as a target in six separate selection campaigns. Each campaign incorporated three rounds of selection from a single library, using one elution condition for phage bound to the immobilized BDDrFVIII. A round of selection consisted of a binding step in which the phage library was first incubated in suspension with the target, followed by a plate wash step to remove unbound or weakly-bound members of the phage library, and an elution step to recover a pool of phage. This results in a population of phage isolates enriched in members that bind the target under binding and wash conditions and release the target under elution conditions. Finally, elution pools from each round of selection were separately amplified by infection and growth in E. coli cultures. The purified phage preparations from this amplification step served as the input for subsequent rounds of selection. The consensus sequence is HpCGSWѲrPCxa/h (uppercase letters stronger than lowercase, Ѳ = V, I, L, F). 3464 FCWVFAFDHCH(14)[24] FCWVFPFNHCS(6)[18] FCWVFPFNHCD(6)[8] FCWVFNFSHCS(3)[24] FCHVFNFVHCS(3)[8] FCWVFPFQHCA(2)[24] FCWVFNWVHCD(1)[14] FCWVFQFRHCH(1)[8] FCWVFPFHHCF(1)[6] 9 Phage display (common panning) 3 PMID:15233528 B-domain deleted form of human factor VIII (BDDrFVIII) Not determined. Not determined. Not determined. TN9 phage display library ELISA Upon completion of the second and third rounds of selection, randomly-chosen individual phage isolates from pools generated by both elution conditions were screened for binding to BDDrFVIII by indirect phage ELISA. Background binding levels were established using wells coated with streptavidin alone or with BSA. Phage isolates exhibiting ELISA signals significantly above background (A630 > 0.25) were scored as positive for binding to BDDrFVIII. The ratio of the ELISA signal with target to the ELISA signal without target was shown. Biotinylated BDDrFVIII protein that was confirmed to be active after derivatization was immobilized onto streptavidin-coated magnetic beads and used as a target in six separate selection campaigns. Each campaign incorporated three rounds of selection from a single library, using one elution condition for phage bound to the immobilized BDDrFVIII. A round of selection consisted of a binding step in which the phage library was first incubated in suspension with the target, followed by a plate wash step to remove unbound or weakly-bound members of the phage library, and an elution step to recover a pool of phage. This results in a population of phage isolates enriched in members that bind the target under binding and wash conditions and release the target under elution conditions. Finally, elution pools from each round of selection were separately amplified by infection and growth in E. coli cultures. The purified phage preparations from this amplification step served as the input for subsequent rounds of selection. The consensus sequence is FCWVFpFxHCx (uppercase letters stronger than lowercase). 3465 SVFPFEVWESLR[1.272, 1.02] YSWHEWYIPQLS[1.185, 0.69] SMPYIAWLALRG[0.929, 0.82] HSTLLNHTTGVL[0.937, 1.06] 4 Phage display (common panning) 3 PMID:30625581 Aflatoxin nanobody Nb28 Aflatoxin B1 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Supernatants containing mimotope-phages were screened by indirect competitive magnetic beads-based ELISA (MB-icELISA) in the presence (10 ng/ mL) or absence of aflatoxin B1 (AFB1). The absorbance was recorded at 450 nm. The maximum absorbance (ODmax) and the 50% inhibitory concentration (IC50, ng/mL) values were determined and shown in the left and right columns of affinity values, respectively. A rapid magnetic beads-based directed competitive ELISA (MB-dcELISA) was developed utilizing Nb28 and its mimotope ME17 (YSWHEWYIPQLS). The 50% inhibitory concentration and the detection limit of the MB-dcELISA were 0.75 and 0.13 ng/mL, respectively, with a linear range of 0.24–2.21 ng/mL. Further validation study indicated good recovery (84.2–116.2%) with low coefficient of variable (2.2%–15.9%) in spiked corn, rice, peanut, feedstuff, corn germ oil and peanut oil samples. 3466 NCKFSGCSVSVCH[5.13] NCRFSGCLQTMCV[8.77] NCRFSGCGTVACV[10.85] NCRFSGCGFKVCV[5.75] NCKFSGCPWELCI[0.78] NCRFSGCVWAKCS[16.90] NCKFSGCQESMCS[1.74] NCRFSGCQWGDCA[23.41] NCKFSGCQFLGCE[2.96] SCKFSGCHRKPCT[1.53] SCRFTFCYYQPCA[111.10] NCRFTFCNLNLCG[29.41] NCRFTFCEFGRCE[>200] NCRFSLCESMACL[71.18] NCKFSACYLTTCY[>200] QCWDRGCENRKCN[1.72] SCLFTFCYFVPCN[>200] TCRQSMCTARTCP[11.56] DCRWSSCTARTCA[8.65] TCGVQACLSARCY[12.9] GCSDQACWSARCV[10.50] NCKFSGCAQLRCY[NA] NCRYSGCFDRPCI[NA] NCRFTLCGLAPCG[NA] NCRYSGCFDRPCI[NA] NCYFSNCGAHSAE[NA] RCLLSACTARACN[NA] GCAQSVCTARYCE[NA] GCSPSRCTARFCP[NA] GCQTSQCTARTCL[NA] RCLPSWCSARTCF[NA] SCPPSRCTARTCP[NA] GCRESSCSARTCP[NA] ACRVSSCTARTCS[NA] ECRASDCSARVCW[NA] WCQASDCTARACI[NA] VCRQSDCSRRTCI[NA] VCRSSDCTKRSCE[NA] PCGEQACYTARCR[NA] LCLSQACWSARCA[NA] GCSAQACASARCL[NA] GCSVQACFSGRCT[NA] GCSQDACYSARCV[NA] ICSVLGCLSARCG[NA] GCGTLACQTARCR[NA] VCVERGCSTARCR[NA] LCADVSCATARCR[NA] GCVRPSCFSARCS[NA] ECQDRSCPFTMCS[NA] 49 Phage display (common panning) 2/3 PMID:24453110 Urokinase-type plasminogen activator Not determined. Not determined. Not determined. XCX4CX4CX phage display library Inhibition assay The inhibition constants Ki (μM) were determined by incubating different concentrations of bicyclic peptide with fixed concentration of human urokinase-type plasminogen activator. The indicated Ki values are averages of at least two measurements. The peptide library was cyclized with the thiol-reactive compound 1,3,5-tris(bromomethyl)benzene (TBMB). The library was then subjected to two or three iterative rounds of phage selection against human urokinase-type plasminogen activator. 3467 RCSGPSCPWQQCS[2.12] RCSGPACVWTQCT[6.88] RCAGPACITQFCT[4.61] RCAGPGCWWAVCG[2.06] RCAGPVCPWTRCG[0.80] RCAGPQCPWSICV[5.39] RCAGPVCPDDPCR[17.73] RCAGPRCPVDMCS[17.73] RCAGAKCPWAVCS[3.76] VCSERGCENRGCG[12.27] LCSDRGCENRWCK[17.73] WCHDRGCENRSCM[14.19] ICLGRGCENRYCG[6.48] TCRQSMCTARTCP[84.92] GCAPTACQSARCG[47.81] CCLGRGCENHRCL[NA] TCQNRGCENRECC[NA] CCRERGCENMVCP[NA] CCGERGCENRACT[NA] QCSINACLSSRCS[NA] GCSVQACLSGRCG[NA] TCGVQACLSARCY[NA] ACRLEACLSARCG[NA] GCTSSACQSARCR[NA] GCGTLACQTARCR[NA] GCGFQACQSARCL[NA] RCASQACYTARCG[NA] SCGVAACHTARCR[NA] VCVRQSCMTARCQ[NA] TCSRASCWTARCL[NA] YCAQASCWTARCG[NA] FCPQVSCLTARCM[NA] SCSQVSCYTARCR[NA] NCAVASCFSARCR[NA] YCVQQSCFSARCV[NA] SCSLQGCLSARCV[NA] RCSIAGCQTARCY[NA] ICRASVCTARTCW[NA] ECGWRACLAARCT[NA] SCSSQACAAARCR[NA] WCASVTCRLYGCM[NA] LCGNYRFSGSMCA[NA] 42 Phage display (common panning) 2/3 PMID:24453110 Urokinase-type plasminogen activator Not determined. Not determined. Not determined. XCX4CX4CX phage display library Inhibition assay The inhibition constants Ki (μM) were determined by incubating different concentrations of bicyclic peptide with fixed concentration of human urokinase-type plasminogen activator. The indicated Ki values are averages of at least two measurements. The peptide library was cyclized with the hydrophilic small molecule 1,3,5-triacryloyl-1,3,5-triazinane (TATA). The library was then subjected to two or three iterative rounds of phage selection against human urokinase-type plasminogen activator. 3468 GCVPQYCPPPLCG[7.33] KCQGSYCPPVACL[3.24] GCQVNYCPPVPCL[1.96] QCQVSYCPPVRCD[8.80] GCAVSYCPPQFCN[41.76] MCHVSYCPWQGCT[12.43] RCMVSYCPEIGCT[10.99] QCSVSYCPQVPCQ[3.54] GCFPSYCPQVACQ[0.78] GCGPSYCPQVACQ[1.77] RCGPSYCPDDFCF[35.90] FCGPSYCPYTGCA[2.98] GCGPSYCSYVPCG[13.09] VCQTSYCPYVPCL[3.68] YCQASYCPGVPCR[9.69] DCASYCPFVECV[84.91] WCQAEYCAFVPCI[4.39] QCQAEYCPRVRCT[1.41] RCQGEYCPPVRCL[6.28] SCQGEYCPFVVCE[6.77] WCQPEYCPQSGCD[103.60] GCQVEYCPPVPCL[15.76] SCQAQYCPEVQCR[4.38] NCQPQYCPRVTCI[1.57] ECQPQYCPSVGCK[20.74] VCQGQYCPPVECS[4.19] GCQVQYCPPVPCL[3.51] ACEVSYCPQVKCQ[NA] SCGPSYCPLVACQ[NA] ECVASYCPQVECL[NA] DCWGSYCQGSYCP[NA] FCDWSYCVLDRCT[NA] GCGSGRCGLTFCM[NA] WCWSGRCGSWGCS[NA] LCPSARCGWQNCE[NA] QCGPLACATARCA[NA] WCGPWGCGTGRCF[NA] WCTRDPCQTGRCY[NA] QCGLDPCQTGRCV[NA] QCRLSACTARSCF[NA] GCFPQYCPMMPCV[NA] DCNRPLCQNAPCL[NA] LCFDQECFWGLCF[NA] WCDWGFCKEQMCN[NA] LFQGACDTGRCA[NA] 45 Phage display (common panning) 2/3 PMID:24453110 Urokinase-type plasminogen activator Not determined. Not determined. Not determined. XCX4CX4CX phage display library Inhibition assay The inhibition constants Ki (μM) were determined by incubating different concentrations of bicyclic peptide with fixed concentration of human urokinase-type plasminogen activator. The indicated Ki values are averages of at least two measurements. The peptide library was cyclized with the hydrophilic small molecule N,N’,N’’-(benzene-1,3,5-triyl)-tris(2-bromoacetamide) (TBAB). The library was then subjected to two or three iterative rounds of phage selection against human urokinase-type plasminogen activator. 3469 RCVQICPSWCG[>200] RCLALCPNWCT[>200] RCVALCPRWCY[>200] RCVKLCPLWCT[NA] RCVDLCPLWCV[NA] RCLAMCPSWCV[NA] 6 Phage display (common panning) 2/3 PMID:24453110 Urokinase-type plasminogen activator Not determined. Not determined. Not determined. XCX3CX3CX phage display library Inhibition assay The inhibition constants Ki (μM) were determined by incubating different concentrations of bicyclic peptide with fixed concentration of human urokinase-type plasminogen activator. The indicated Ki values are averages of at least two measurements. The peptide library was cyclized with the thiol-reactive compound 1,3,5-tris(bromomethyl)benzene (TBMB). The library was then subjected to two or three iterative rounds of phage selection against human urokinase-type plasminogen activator. 3470 RCLVPCRYLCW[>200] RCLLPCRYWCE[>200] RCVRPCYFPCA[>200] GCSWWCNRACP[>200] VCLMICHYPCL[>200] RCLVPCRLHCW[NA] RCLLPCRYWCP[NA] RCLVPCKYWCY[NA] RCLLPCNLWCF[NA] RCLGPCWIVCP[NA] RCVRPCFIPCA[NA] RCVRPCPWTCS[NA] RCVAPCWWGCN[NA] RCQYWCNRACL[NA] TCSWWCNRSCL[NA] GCSWWCNRACL[NA] LCYQICHYPCS[NA] TCFFICHYPCL[NA] 18 Phage display (common panning) 2/3 PMID:24453110 Urokinase-type plasminogen activator Not determined. Not determined. Not determined. XCX3CX3CX phage display library Inhibition assay The inhibition constants Ki (μM) were determined by incubating different concentrations of bicyclic peptide with fixed concentration of human urokinase-type plasminogen activator. The indicated Ki values are averages of at least two measurements. The peptide library was cyclized with the hydrophilic small molecule 1,3,5-triacryloyl-1,3,5-triazinane (TATA). The library was then subjected to two or three iterative rounds of phage selection against human urokinase-type plasminogen activator. 3471 VCNQICGRLCE[0.30] VCNQVCGRQCD[0.36] RCNYICGRDCS[0.37] SCNDVCGRDCT[1.48] SCNQICGRWCF[NA] TCNQICGRSCP[NA] TCNQICGRICV[NA] NCNQVCGRECG[NA] VCNQVCGRLCP[NA] LCNQVCGRQCL[NA] KCNQVCGRLCA[NA] LCNSLCGRICD[NA] WCNSICGRSCF[NA] KCNSICGRNCS[NA] VCNSVCGRLCS[NA] RCNDICGRHCL[NA] RCNAVCGRDCN[NA] SCNAICGRACD[NA] GCNRICGRLCE[NA] 19 Phage display (common panning) 2/3 PMID:24453110 Urokinase-type plasminogen activator Not determined. Not determined. Not determined. XCX3CX3CX phage display library Inhibition assay The inhibition constants Ki (μM) were determined by incubating different concentrations of bicyclic peptide with fixed concentration of human urokinase-type plasminogen activator. The indicated Ki values are averages of at least two measurements. The peptide library was cyclized with the hydrophilic small molecule N,N’,N’’-(benzene-1,3,5-triyl)-tris(2-bromoacetamide) (TBAB). The library was then subjected to two or three iterative rounds of phage selection against human urokinase-type plasminogen activator. 3472 CTGTPARQC(4/47)[15.66 ± 2.82] CKNSMFATC(4/47)[10.56 ± 1.09] CTNKHSPKC(3/47)[10.61 ± 0.73] CSPKNILHC(3/47)[NA] CFPSPTRTC(2/47)[NA] CNAGTLGRC(2/47)[NA] CNKEFASQC(2/47)[NA] CNSANNRIC(2/47)[NA] CSTQSTTSC(2/47)[NA] CAPPGKSEC(1/47)[NA] CAPKNILHC(1/47)[NA] CAPSSSATC(1/47)[NA] CESKTPKNC(1/47)[NA] CFDHHTNSC(1/47)[NA] CGTRETLSC(1/47)[NA] CHAALNRSC(1/47)[NA] CHHSNQRQC(1/47)[NA] CHRPDTRSC(1/47)[NA] CNRESPHLC(1/47)[NA] CNSRSHAIC(1/47)[NA] CPMPSTSYC(1/47)[NA] CPVTSRSDC(1/47)[NA] CRAWNEAPC(1/47)[NA] CSDRGLPSC(1/47)[NA] CSSKSDHSC(1/47)[NA] CTAYPAKAC(1/47)[NA] CTGAPARWC(1/47)[NA] CTKTGLHIC(1/47)[NA] CTSTAPLKC(1/47)[NA] CVTSPFHNC(1/47)[NA] CYSPRGGSC(1/47)[NA] CYTNPDNVC(1/47)[NA] 32 Phage display (subtractive panning, in vivo) 4 PMID:30788426 Human prostate cancer cell line LNCaP Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Binding assay LN1 (CTGTPARQC), LN2 (CKNSMFATC), and LN3 (CTNKHSPKC) peptides modified with a biotin molecule were incubated with LNCaP cells for 24 h at a concentration of 1 mg/mL and the binding affinities of the respective peptides were compared using fluorescence with Avidin D-conjugated TRITC (biotinylated peptides are stained red). The fluorescence intensity of each target peptide bound to LNCaP cells (n = 6 per group) was reporduced from Figure 2C and shown. The phage library was injected into C57BL/6 mice through the tail vein at a titer of 1.0e9 plaque-forming units (PFU)/mL in 100 mL Tris-buffered saline (50 mM Tris-HCl [pH7.5]). Five minutes after the injection, blood was recovered from the heart in order to remove non-specifically bound phages. The collected blood was dissolved in DMEM (Thermo Fisher Scientific) containing protease inhibitor (Sigma-Aldrich, St Louis, MO, USA). The solution was introduced into Escherichia coli ER2738 (New England Biolabs), and the number of phages in the blood was titrated by plaque-forming units. After amplification to 1.0e11 PFU, the phages were injected into new C57BL/6 mice. After three rounds of in vivo panning with mice, the phages were injected into the femoral vein of a 15-year-old female cynomolgus monkey. Five minutes after the injection, blood was taken from the femoral vein and dissolved in DMEM with protease inhibitor. The solution was centrifuged to collect the serum, from which phages were amplified to 1.0e11 PFU. These phages were injected into the tail vein of a C.B-17 SCID mouse with tumor xenografts (human prostate cancer cell line LNCaP (RCB2144)), and xenografted prostate cancer tissue was isolated after trans-cardiac removal of blood. Phages were recovered from the homogenized tissue and amplified as described above. Three additional cycles of in vivo phage panning were performed. After four cycles of in vivo panning in C.B-17 SCID mice, phage DNA was isolated from selected phage plaques with high affinity to prostate cancer tissue. LN1 (CTGTPARQC), LN2 (CKNSMFATC), and LN3 (CTNKHSPKC) were synthesized and evaluated for binding and biological activity. LN1 showed the highest avidity for LNCaP prostate cancer cells in vitro and was thus administered to tumor-bearing mice to evaluate in vivo binding. Strikingly, LN1 specifically bound to the tumor tissue and exhibited very low reactivity with normal liver and kidney tissues. To demonstrate that LN1 could specifically deliver drugs to prostate cancer tissue, a therapeutic peptide, LN1-KLA (C-TGTPARQ-CGGG-D[KLAKLAK]2), was prepared and used to treat LNCaP cells in vitro and was also administered to tumor-bearing mice. The therapeutic peptide significantly suppressed growth of the cells both in vitro and in vivo. 3473 ANTELALANRKH(2/110) NYLPHQSSSPSR(2/110) SLPNLPPTYAKP(2/110) ASNHSIPTFPLK(2/110) NPMNNVAQNPGP(2/110) TLGLRPVPVATT(2/110) GSWNTFRAQPTI(2/110) SQALSTSRQDLR(2/110) HSACLGPSNLQC(2/110) RVQPAHFNVMGQ(1/110) MVGTADGTLLDP(1/110) TMHHAAIAHPPH(1/110) GIVTNQHDSNAN(1/110) GLTFQVPWHANM(1/110) GHPMMPPKSEIR(1/110) TMAQGVAQRYGN(1/110) SHQPGDQSPANN(1/110) DLINIDRNHSFR(1/110) LPKQCSLLTSAC(1/110) NFTLQAHPHKYP(1/110) STDHGSWQKSRA(1/110) VPQLHHLMPHFD(1/110) TSMSQHFHVHRL(1/110) SPLTPPHAPETH(1/110) CPTDVRSGCMGT(1/110) IEMTRTNLNDVN(1/110) HTQHIQSDDHLA(1/110) NDLQRHRLTAGP(1/110) DDTQNSQNMDTL(1/110) 29 Phage display (common panning) 3 PMID:31040030 Gallium ion Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Target preparation was carried out by immobilization of gallium ions on small monolithic ion exchange columns (CIM Disk Monolithic Column, BIA Separations d.o.o., Ajdovscina, Slovenia) for Äkta avant 25 FPLC system (GE Healthcare Europe GmbH, Freiburg,Germany). Original phage library or subsequently enriched phage pools were diluted in the respective buffer to a final volume of 1 ml and applied in a repetitive recycling loop of 15 ml. Unbound and rather unspecific binding phage were removed by washing with column volumes (CV) of the buffer at a flow rate of 1 ml/min. Good binding phage were eluted by applying 40 CV of the eluent at a flow rate of 1 ml/min. The eluate was collected in fractions, 1 ml each. In a final step gallium together with the remaining tight bound phage were stripped by applying 40 CV 1 M HCl and fractionated as well. Phage clones expressing the peptide sequences TMHHAAIAHPPH, SQALSTSRQDLR and HTQHIQSDDHLA were characterized to bind >10 fold better to a target that presents immobilized gallium ions than control phage, displaying no peptide sequence. 3474 CIAVPSNLC CKTPNGHLC CKXPKGHLC 3 Phage display (common panning) 3 PMID:30850705 Human colorectal cancer cell line RKO Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) An adaptation of the BRASIL method using RKO cells as target was used. A total of three rounds of selection with human epithelial CRC cell line RKO (ATCC CRL 2577) were performed through in vitro biopanning. In each round, the phages that specifically bound to target cells were recovered and used for the next round of selection. In the three rounds of selection, the obtained phage pool was not amplified between rounds. 3475 CRPTYSPSC CKIHSSETC CPKSNNGVC CIGNSNTLC 4 Phage display (common panning) 4 PMID:30850705 Human colorectal cancer cell line RKO Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) An adaptation of the BRASIL method using RKO cells as target was used. A total of four rounds of selection with human epithelial CRC cell line RKO (ATCC CRL 2577) were performed through in vitro biopanning. In each round, the phages that specifically bound to target cells were recovered and used for the next round of selection. In the three initial rounds of selection, the obtained phage pool was not amplified between rounds. 3476 CPKSNNGVC[0.27 ± 0.04] CKTPNGHLC[0.17 ± 0.01] CQSISTAHC[0.18 ± 0.03] CNDDVPNKC[0.17 ± 0.02] CEIPGKVVC[0.17 ± 0.02] CLRTPANHC[0.20 ± 0.02] 6 Phage display (subtractive panning) 4 PMID:30850705 Human colorectal cancer cell line RKO Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Assessment by cell-ELISA of the binding selectivity to RKO of six phage clones was performed. The plate was read at 562 nm on an automated ELISA plate reader (Biotech Synergy HT). The M13KE wild-type phage was used as a negative control. The OD562 values were reproduced from Figure 1. The OD562 value of the M13KE wild-type phage is 0.05 ± 0.01. An adaptation of the BRASIL method using RKO cells as target was used. A total of four rounds of selection with human epithelial CRC cell line RKO (ATCC CRL 2577) were performed through in vitro biopanning, followed by a negative selection step against normal colon CCD-841-CoN cell line. In each round, the phages that specifically bound to target cells were recovered and used for the next round of selection. In the three initial rounds of selection, the obtained phage pool was not amplified between rounds. After four rounds of selection plus a negative step with normal colorectal cells, CCD-841-CoN, there was an obvious phage enrichment that specifically bound to RKO cells. Cell-based enzyme-linked immunosorbent assay (ELISA) was performed to assess the most specific peptides leading to the selection of the peptide sequence CPKSNNGVC. Through fluorescence microscopy and cytometry, the synthetic peptide RKOpep was shown to specifically bind to RKO cells, as well as to other human colorectal cancer cells including Caco-2, HCT 116 and HCT-15, but not to the normal non-cancer cells. Moreover, it was shown that RKOpep specifically targeted human colorectal cancer cell tissues. A bioinformatics analysis suggested that the RKOpep targets the monocarboxylate transporter 1, which has been implicated in colorectal cancer progression and prognosis, proven through gene knockdown approaches and shown by immunocytochemistry co-localization studies. The peptide herein identified can be a potential candidate for targeted therapies for colorectal cancer. 3477 QATHRSH 1 Phage display (subtractive panning, competitive panning) 3 PMID:30796964 Aromatic-L-amino-acid decarboxylase (EC:4.1.1.28), AADC Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Two petri dishes were coated with 10 μg/ml of either MBP (maltose binding protein) or MBP-DDC in 0.1 Μ NaHCO3, pH 8.6, overnight at 4°C. After removal of the residual protein solution, non-specific binding sites of the petri dish were blocked with 5 mg/ml bovine serum albumin) and the petri dishes were incubated at 4°C for 1 hr. All remaining procedures were performed at room temperature. The blocking buffer was removed from each petri dish and subsequently the dishes were washed 6 times with TBST (Tris buffered saline, 0.1% Tween 20, pH 7.4). 2x1011 library phages were added to the petri dish coated with ΜΒΡ and incubated for 1 hr. The phages interacting with MBP or BSA were bound to the petri dish (non-specific interaction). This first incubation resulted in removing the phages that were bound to the fusion protein by interacting with MBP, thus allowing that in the next step only the phages interacting specifically with DDC would bind. The supernatant that contained the phages not bound to MBP or BSA was incubated for 1 hr with the petri dish coated with the fusion protein. Following incubation and removal of the supernatant, 10 washes with TBST were performed. Elution of the phages was performed with a 1 hr incubation with shaking with free MBP-DDC (~100 μg/ml in TBS) or with L-Dopa (~100 μg/ml in TBS). For the (His)6DDC fusion protein the above technique was used, but with agarose magnetic beads. Human phosphatidyl-inositol-3-kinase (GenBank accession number AAG61115.1) contains the peptide ATHRS at positions 127–131, showing high similarity with the heptapeptide QATHRSH, isolated by the phage display method ten times in total, eluted both with MBP-DDC (eight times) and L-Dopa (two times). 3478 HSVEDVS(14/50)[1.35] PGVAERS(10/50)[2.58] LPRSHPI(7/50)[5.25] SSIWYSP(6/50)[2.61] TFHSTFS(5/50)[10.23] VAVSNTH(4/50)[3.20] 6 Phage display (subtractive panning) 4 PMID:30986050 D2-MT1-MMP (MT-loop deleted MMP-14) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding affinities of the top six high ratio phage peptides were determined by ELISA. The resulting Kd value (nM) is a dissociation constant, representing the proportion of dissociated molecules to initial molecule number in unit time, indicating that the higher the Kd value, the lower the affinity, and vice versa. In each round of panning, one group of WT-MT1-MMP and three groups of D2-MT1-MMP (2 mL each) were coated onto a 35 mm sterile polystyrene dish to make sure that the surface is completely wet. The plates were then kept in a humidified incubator at 4 °C overnight and blocked using a blocking buffer (0.1 M NaHCO3, pH = 9.6, 5 mg/mL BSA) for 2 h at room temperature (RT) to avoid nonspecific binding. After discarding the blocking solution and washing six times with TBST (TBS containing 0.1% [v/v] Tween-20), 10 μL of Ph.D. 7-peptide library (2.0e11 pfu) was diluted in 1 mL of TBST and exposed to the No.1 D2-MT1-MMP plate for 1 h at RT with gentle shaking. Then, the unbound phages were transferred to No.2 D2-MT1-MMP plate for 1 h at RT under gentle rotation. This procedure was repeated for No.3 D2-MT1-MMP plate. After these three steps, phage peptide which did not combination with D2-MT1-MMP was obtained and placed in No.4 plate, which is WT-MT1-MMP. After 1 h incubation, the bound phages were collected by adding 1 mL of 0.2 M Glycine-HCl (pH = 2.2) containing 1 mg/mL BSA to the plate for 8 min and neutralized with 150 μL of 1 M Tris-HCl (pH = 9.1). Then, 1 μL of phages was collected for phage tittering, and the remaining phages were mixed with 200 μL of ER2738 culture (at early log stage) for further amplification. After bacteriolysis, by centrifugation and PEG precipitation, the phages were recovered from the culture supernatant, and then dissolved in TBS. The titer was determined on LB/IPTG/Xgal plates and used as input phages for the next cycle. After four cycles of panning, the phage clones that had higher binding affinity to MT-loop were obtained. Several clones were randomly selected and −96 gIII sequencing primer was used to obtain the peptide sequences. In this study, we considered the 3-dimensional (3-D) conformation of the MT-loop area in the MT1-MMP molecule and designed a novel strategy to screen the Ph.D. peptide library. The peptide HSVEDVS (HS7) we obtained showed a better binding affinity to WT-MT1-MMP than the peptide HWKHLHNTKTFL (AF7p) as observed through enzyme-linked immunosorbent assay (ELISA) and biolayer interferometry (BLI). The new peptide labeled and attached MT1-MMP expression cell lines HT1080 and did not show any toxicity to cells. Furthermore, for in vivo imaging, HT1080 tumor-bearing mice with higher MT1-MMP expression accumulated more Cy5.5-HS7 than mice with MT1-MMP low-expression cell lines A549 at tumor sites, and the half-life of HS7 was longer than that of AF7p, as confirmed by ex vivo imaging of the main organs. These results suggest the feasibility of using the subtraction biopanning strategy to screen the affinity peptide targeting MT-loop regions and HS7 is a superior probe for noninvasively imaging MT1-MMP expression in MT1-MMP-positive tumor models. It provides impetus for further studies to use HS7 in early diagnosis of tumors and in peptide-mediated drugs. 3479 CNAGHLSQC(2)[0.67 ± 0.03] CSLNHTVNC(1)[0.86] CSAKTTSAC(1)[0.63] 3 Phage display (subtractive panning, competitive panning) 3 PMID:30968602 Fibroblast growth factor receptor 1, FGFR-1 Fibroblast growth factor 1, FGF-1 1EVT, Not determined. Ph.D.-C7C phage display library (CX7C) ELISA ELISA was conducted after the third round of selection. The absorbance was measured at 450 nm and reproduced from Figure 1A. After the second and third rounds of biopanning, additional counterselection with Fc fragment was performed. Additionally after the last round of selection, binding phages were eluted with 100 molar excess of fibroblast growth factor 1 (FGF1) over applied phage library. We used the phage display technique to select cyclic peptides F8 (CSLNHTVNC) and G10 (CSAKTTSAC) as binders of the fibroblast growth factor 1 (FGF1)–FGFR1 interface. ELISA and in vitro cell assays were performed to reveal that cyclic peptide F8 is more effective in preventing the FGF1–FGFR1 interaction, and also decreases FGF1-induced proliferation of BA/F3 FGFR1c cells by over 40%. Such an effect was not observed for BA/F3 cells lacking FGFR1. Therefore, cyclic peptide F8 can act as a FGF1–FGFR1 interaction antagonist, and may be suitable for further development for potential use in therapies against FGFR1-expressing cancer cells. 3480 HPSALMKPTSHA(4)[6.12 ± 0.24] DRWVARDPASIF(4)[7.48 ± 0.35] NPHAPSSFYEAY(3)[5.47 ± 0.10] TVGLPMTYYMHT(3)[8.23 ± 0.30] TSSPLTRWSSSL(2)[5.51 ± 0.16] AWADQPVTAPNR(2)[4.54 ± 0.05] HLTTTHPEPPYG(2)[4.17 ± 0.16] IPLGRDGGSYQR(2)[3.57 ± 0.13] MDENVATNQLMI(2)[5.87 ± 0.48] ALGDSCRYCRLL(1)[NA] ATEERSRIWMFL(1)[NA] CVASARGAQIGM(1)[NA] DDFRVWWPNFPR(1)[NA] DPVGLGGWWAKV(1)[NA] DSSQWDKIYSWT(1)[NA] GFAVGARDSLMF(1)[NA] GSAPLLTVDTSK(1)[NA] HSKAFPVLYPLR(1)[NA] LGHSGGPTRPSW(1)[NA] MLDQRPMSSYAG(1)[NA] MSLDSFRVDRRA(1)[NA] QGMVAESYSPLS(1)[NA] SALKGLFPADHH(1)[NA] SLECDELIHSQI(1)[NA] STLGFNPPAILP(1)[NA] STPGCCAHDHFR(1)[NA] SVPMGSLASLES(1)[NA] TAHASLDDQGLR(1)[NA] TPQSFWQKGSLV(1)[NA] VSGQRSVGTPLS(1)[NA] WDFRQWWQPSGG(1)[NA] 31 Phage display (common panning) 4 PMID:30937184 Proheparin-binding EGF-like growth factor Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Quantification of binding abilities of peptides was performed by using enzyme-linked immunosorbent assay (ELISA). Absorbance was measured at 405 nm. The relative binding abilities were calculated using the following equation: Absorbance when sHB-EGF coated/Absorbance when bovine serum albumin (BSA) coated. The relative binding values were reproduced from Figure 3B and shown. The relative binding value of the wild type M13 phage is 2.6079 ± 0.14361. Six hundred microliters of sHB-EGF (100 μg/ml in NaHCO3, pH 8.6) was coated on polystyrene 6-well cell culture plate overnight at 4 °C. Other steps were performed according to manual of Ph.D-12. In order to avoid nonspecific binding, the phage elusion in each round was incubated with a new plate for 20 min at room temperature. After 4 rounds of biopanning, the isolated colonies of E. coli ER2738 were cultured and the remained phages were sequenced. Two peptides, no. 7 (HLTTTHPEPPYG) and no. 29 (TPQSFWQKGSLV) were found mildly binding to HB-EGF. Then the effects of these peptides on HB-EGF functions were examined and both peptides no. 7 and no. 29 were found indeed inhibiting the functions of HB-EGF in promoting migration and invasion of SKOV3 and HO-8910 cells in vitro. Further mechanism investigation showed that peptides no. 7 and no. 29 inhibited HB-EGF-promoted cell migration and invasion through attenuating activation of the EGFR signaling pathway manifested by decreased p-Erk1/2 and Snail levels. More importantly, peptides no. 7 and no. 29 showed strong activities in inhibiting migration of SKOV3 cells in vivo. 3481 ERTYSPSTAVRS[1.458 ± 0.28] PSTAVR[NA] SPSTAVRS[NA] TYSPSTAVRSAA[NA] PSTETR[NA] LPSTETRHV[NA] YSPSTETRSA[NA] 7 Phage display (subtractive panning) 3 PMID:31132884 The catalytic domain of rho-associated protein kinase 1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The strength of interactions between Peptide7 and ROCK1 and ROCK2 was then alyzed by performing an SPR study. The binding of Peptide7 to ROCK1 as compared to ROCK2 demonstrated small differences even though bith proteins have 92% similarity in sequence in the catalytic domain. Upon inspection, there are only 3 amino acid differences in the activation loop. The average of three experiments yielded KD values of 1.458 ± 0.28 µM for ROCK1 (1–553) and 5.153 ± 1.15 µM for ROCK2. Phage particles were first mixed with MBP protein to avoid the negative selection by elimiting phages that could bind to MBP. The supertant which contained only non-MBP-binding phages was then incubated with MBP-ROCK1 (5–348). Peptide7(ERTYSPSTAVRS), a promising ROCK inhibitory peptide for both ROCK isoforms, measured at 1.45 ± 0.28 µM for ROCK1 (1–553) and 5.15 ± 1.15 µM for ROCK2. Peptide7 reduced cellular migration in wound healing assays. The binding epitope on ROCK1 was mapped to the flexible activation loop within the catalytic domain. Peptide alanine scanning mutants helped identify critical amino acids to generate optimized Peptide22. This compact ROCK inhibitor facilitated vascular relaxation, blocked neovascularization of endothelial cells, and inhibited MLC phosphatase phosphorylation. 3482 CSLTQDQGC(328,760) CADAQADVC(140,578) CKASRLGRC(139,938) CRAKGRDAC(103,044) CVGRPDA*C(84,760) CSRTEGDVC(67,529) C*SSLVKTC(61,014) CKRSS*YFC(57,309) CRNGEGAQC(54,832) CGEL*CNDC(54,226) 10 1 PMID:32035108 Inflamed central nervous system (CNS) of mice with experimental autoimmune encephalomyelitis (EAE) Not determined. Not determined. Not determined. CX7C T7 phage display library A tissue homogete equivalent to 100 mg of tissue was incubated with phage library at 4 °C for 1 h. Thereafter, the tissue homogete was washed to remove the unbound phages, whereas the bound-phages were rescued using BLT5615 strain of E. coli. The phages were then amplified and phages were injected intravenously (i.v.) into EAE mice that had clinical score between 2 and 3. The phages were circulated for 15 min. Thereafter, animals were anesthetized, perfused with saline, and CNS tissue was collected and homogenized. MS-1 (amino acid sequence CRGGKRSSC), was shown to selectively recognize CNS tissue in EAE mice. Individually cloned phages with this insert preferentially homed to EAE CNS after an intravenous injection. Similarly, systemically-administered fluorescence-labeled synthetic MS-1 peptide showed selective accumulation in the spil cord of EAE mice. 3483 CSLTQDQGC CRAKGRDAC CAMGNGGDC CTTPAKNNC CTEQIEERC CKASRLGRC CVGRPDA*C C*SSLVKTC CKRSS*YFC SLTQDQG, RAKGRDA, AMGNGGD, RPGESS, PGTVKR, KRSS, GDRLV, TTPAKNN and TEQIEER 9 3 PMID:32035108 Inflamed central nervous system (CNS) of mice with experimental autoimmune encephalomyelitis (EAE) Not determined. Not determined. Not determined. CX7C T7 phage display library A tissue homogete equivalent to 100 mg of tissue was incubated with phage library at 4 °C for 1 h. Thereafter, the tissue homogete was washed to remove the unbound phages, whereas the bound-phages were rescued using BLT5615 strain of E. coli. The phages were then amplified and phages were injected intravenously (i.v.) into EAE mice that had clinical score between 2 and 3. The phages were circulated for 15 min. Thereafter, animals were anesthetized, perfused with saline, and CNS tissue was collected and homogenized. MS-1 (amino acid sequence CRGGKRSSC), was shown to selectively recognize CNS tissue in EAE mice. Individually cloned phages with this insert preferentially homed to EAE CNS after an intravenous injection. Similarly, systemically-administered fluorescence-labeled synthetic MS-1 peptide showed selective accumulation in the spil cord of EAE mice. 3484 YYVSWPPDMMHY(11/17)[500 ± 25 nM, 299.3 nM] GRTPLLDSFLAG(3/17)[~43 μM, NA] TQQDYAWLLDH(1/17)[NA] SYSNFPLLHSYW(1/17)[NA] HDTWPLLATKTS(1/17)[NA] [PL]LL[DAH] 5 Phage display (common panning) 3 PMID:32659860 Signal transducer and activator of transcription 3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The affinity (Kd) of the peptides to STAT3 was determined by isothermal titration calorimetry (ITC) using no ITC calorimeter (TA Instruments, DE, USA). The alysis of which shows that peptide 2 interacted with higher affinity (Kd = 500 ± 25 nM) while peptide 1 was a slightly weaker (Kd = 1.79 ± 0.12 μM). Peptide 3, however, showed nearly 80x weaker compared to peptide 2 (Kd ~ 43 μM). All the three peptides were found to bind with a release of heat. For peptide ELISA, the strength of the association was recorded by Cytation 3 Imaging reader (BioTek Instruments). The result shows that the peptides interacted with STAT3 in a dose‐dependent manner. Peptide 1 was shown to have an affinity of 299.3 nM while the value for peptide 2 is 232.9 nM. Peptide 3 did not reach saturation under the identical conditions, suggesting a weaker interaction with STAT3. The first column of the affinity values is the Kd value determined by ITC. The second column is the strength of the association determined by peptide ELISA. A 100‐fold dilution of the PhD‐12™ library (New England Biolabs) was then added and incubated for 1 hr.The precipitated phages were pelleted by centrifugation (60 min, 16,100 g, 4°C), and the tubes dried for 20 min at room temperature (RT). The pellets were then re‐suspended in 1 ml of TBST buffer containing 0.02% w/v N3 and 1% (w/v) BSA. After re‐precipitation and suspension, the top 80% of the supertant was used for the next round of phage amplifications and selection. The YYVSWPPDMMHY sequence was found to be enriched by 36% while another with a short consensus motif was displayed in 20% of the phages. Binding alysis by isothermal titration calorimetry shows the most displayed peptide interacted with a Kd of 1.79 μM, which on modification of its structure to mimic the tural binding partners of STAT3 brought the affinity to high nomolar range (Kd = 500 nM). Using a panel of tumor cell lines, we show that the peptides prevented the proliferation of triple‐negative breast cancer cells with a moderate activity (GI50 = 50 μM). 3485 AHLPTSMLKGQG(12)[0.0224] YHFNGCEDPLCR(2)[0.0712] KVWSLVNQGGQF(2)[3.13] GHIPTCLTPMCR(1)[0.137] HSIRDGFRSTPV(1)[0.466] TGQTVTGLSYIF(1)[2.32] KVVSLSALQSMT(1)[0.665] WTSQPHLQHVDD(1)[5.29] GHLAVNMPRASL(1)[0.257] HHTGCLSPLSCS(1)[0.00257] HWKVTTWNSSTV(1)[3.52] SGVYKVAYDWQH(1)[5.00] HPWCCGLRLDLR(1)[0.0873] GH,PT,QGGQ,KV 13 Phage display (common panning) 3 PMID:32664518 Extracellular domain of low-density lipoprotein (LDL) receptor Low-density lipoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The apparent dissociation constant (K*d, M) was determined by ELISA. Each clone was incubated with LDLR and BSA-coated wells at a concentration of 5x1011 phages in 100µl of buffer for 2 hours under stirring (350 rpm). The blank was incubated with the buffer. Then the HRP-conjugated monoclol anti-M13 antibody (Amersham Pharmacia Biotech Benelux, Roosendaal, The Netherlands) was incubated for 1 hour (dilution 1:5000 in TBSC with 0.5% BSA) to detect bound phages. The phage displayed random library of linear dodecapeptides (Ph.D.-12, New England Biolabs Inc., Bioké, Leiden, The Netherlands) was screened against the extracellular domain (ED) of LDLR.Three rounds of selection were performed to obtain a pool of phages with an increasing affinity to the target. Fifty clones were isolated from this 3rd pool of phages, and their binding to the ED-LDLR and the BSA employed as a control protein was evaluated at one concentration. The peptide (HPWCCGLRLDLR) was able to bind the ED-LDLR in the presence of tural ligands and dissociated at acidic pH and in the absence of calcium, in a similar manner as the LDL. In vitro, The peptide was endocytosed by endothelial cells through the caveolae-dependent pathway, proper to the LDLR route in BBB, suggesting the prevention of its lysosomal degradation. The in vivo studies performed by magnetic resonce imaging and fluorescent lifetime imaging suggested the brain penetration of this ED-LDLR-targeted peptide. 3486 APTTHMWLNYWK(25/39) THWMXNY 1 Phage display (common panning) PMID:31778678 Anti-D-antigen monoclonal antibody 1G10 D-antigen Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Three phage clones were selected because of their high frequency in bio-panning, good affinity and specificity for binding to mAb 1G10 in phage ELISA , as well as their similarity with the epitope of PV-I Sabin antigen performed in phage competitive ELISA. Phage clones were selected by solution-phase panning with protein G magnetic beads (NEB) that exhibited high affinity for IgG. 3487 LPFMPWI(3/17) XPF 1 Phage display (common panning) PMID:31778678 Anti-D-antigen monoclonal antibody 1G10 D-antigen Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Three phage clones were selected because of their high frequency in bio-panning, good affinity and specificity for binding to mAb 1G11 in phage ELISA , as well as their similarity with the epitope of PV-I Sabin antigen performed in phage competitive ELISA_x0000_ Phage clones were selected by solution-phase panning with protein G magnetic beads (NEB) that exhibited high affinity for IgG. 3488 CSLTQYRYC(1/15) XTQ 1 Phage display (common panning) PMID:31778678 Anti-D-antigen monoclonal antibody 1G10 D-antigen Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Three phage clones were selected because of their high frequency in bio-panning, good affinity and specificity for binding to mAb 1G12 in phage ELISA , as well as their similarity with the epitope of PV-I Sabin antigen performed in phage competitive ELISA_x0000_ Phage clones were selected by solution-phase panning with protein G magnetic beads (NEB) that exhibited high affinity for IgG. 3489 LTPHKHHKHLHA(19/33) GPHHMHHHRTHH(7/33) WPRHHWHTNYMR(1/33) GWHSPHAHWRVK(1/33) THYNPLRINPIT(1/33) KVHIMHFHHHSL(1/33) HSWSTIKRIETM(1/33) WPHLQHHKATSR(1/33) HDRMTKSSFSPP(1/33) 9 Phage display (common panning) 3 PMID:32568515 Se0 nanoparticle (SeNP) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Phage binding to SeNPs proceeded for 1 hour in selection rounds 1 and 2 but was reduced to 10 minutes in round 3. After each selection round, bound phage were acid-eluted with a Glycine-HCl (pH 2.2) based elution buffer for 20 minutes. A selenium noparticle binding peptide (LTPHKHHKHLHA) was genetically fused to a metalloid reductase that reduces selenite to a Se0 noparticle (SeNP) form. The fusion of the Se binding peptide to the metalloid reductase regulates the size of the resulting SeNP to ~35 nm average diameter, where without the peptide, SeNPs grow to micron sized polydisperse precipitates. The SeNP product remains associated with the enzyme/peptide fusion. 3490 SGVYKVAYDWQH(13/42) VHWDFRQWWQPS(10/42) HHWNSLWWPWKT(9/42) HFWHWPLWTRDH(5/42) HHYWWPWQITNS(5/42) TWWHWPRMYMGL(1/15) WHWWPHWDTGRA(1/15) WYPYNWTHWFTH(1/15) HLWYPWIWPMQQ(1/15) WHWNAWNWSSQQ(1/15) WHWSWLVHPHTL(1/15) LTNNLHKQVGPW(1/15) WHFTWWVDNRMT(1/15) HFMSYAGASTWA(1/15) GLHTSATNLYLH(1/15) GSAPLLTVDTSK(1/15) WEYSVVTPVDFL(1/15) GLTSQRLDGLPP(1/15) NPHIAHSLHSWV(1/15) HPHDYNDLTSPF(1/15) 20 Phage display (common panning) 3 PMID:32413370 Anti-HIV-1 monoclonal antibody Y498 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA A phage ELISA was performed to determine the binding 12 mer-peptides. The results were measured at a wavelength of 450 nm. A random phage display peptide library (PhD-12) was purchased from New England Biolabs.After three rounds of selection, the selected phage clones were identified by single-strand D sequencing and phage ELISA, and the encoded peptide sequences were deduced. The concentration of washing buffer was increased gradually each round to a maximum of 0.5%TBST. To identify the key amino acid recognition site contacted with neutralizing antibody Y498, peptides were panned from the PhD-12 peptide library and predicted using online software. Then, four key amino acid sites, G367, D368, E370, and V372 located on the CD4 binding loop on gp120 of envelope of human immunodeficiency virus-1 (HIV-1), were found to determine the neutralization of antibody Y498. Residue E370 is in the deep part of the CD4 binding loop, which affects Y498-mediated neutralization. 3491 CHMRQGMAC(33/50)[1.40139] CSYPDTHRC(2/50)[0.3324] CQHPGSTMC(2/50)[0.31154] CTATESTVC(2/50)[0.38348] CNDWLSNSC(2/50)[0.49799] CQQGLSSQC(1/50)[0.79257] CHGPRASQC(1/50)[0.39923] CKQTTSSSC(1/50)[0.78065] CPLAYPHTC(1/50)[0.77129] CKETWWPHC(1/50)[1.19017] CMHTQTPWC(1/50)[0.34602] CTTHTYFGC(1/50)[0.63293] CGLKHTLKC(1/50)[0.48309] CLARNATWC(1/50)[0.50906] 14 Phage display (subtractive panning) 4 PMID:31247207 Plasma from 10 Alzheimer's disease patients Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The absorbance value at 450 nm was determined with a microplate reader. Data (OD450) shown were reproduced from Figure 2C in the reference. In the first round of selection, phage library was selected against the pooled Alzheimer's disease (AD) plasma with albumin/IgG depletion. The second round was negative selection against pooled plasma from 10 Controls (5 male and 5 female) without albumin/IgG depletion. The third and fourth rounds of selection were against the same pooled AD plasma with albumin/IgG depletion. We further characterized one AD-specific peptide (AD#1 peptide, CHMRQGMAC) and one control-specific peptide (Con#1 peptide, CDGARHGRC), and evaluated their diagnostic performance in independent validation set (35 AD patients, 45 MCI, 45 controls and 20 PD patients). Our results show that both AD#1 peptide and Con#1 peptide could distinguish AD/MCI patients from controls and combition of these two peptides could greatly improve the diagnostic performance (AUC is above 0.80 in ROC curve alysis). In addition, we found that AD#1 peptide stained Aβ-treated primary astrocyte and bound to recombint human YKL-40 protein in in-vitro assay. It supports that AD#1 peptide detects AD inflammation related cytokine. 3492 CDGARHGRC(30/50)[1.63760] CQSPKTPFC(3/50)[1.08744] CSNGSDSSC(2/50)[1.13536] CTSVPHRYC(2/50)[0.40443] CTSTGRSNC(2/50)[0.74506] CSVSQLKLC(2/50)[0.54469] CTPLSYKPC(2/50)[0.40094] CLGQYHQRC(1/50)[1.18241] CLPSGNNRC(1/50)[0.53075] CKHTTSTSC(1/50)[0.36385] CWKHGNFNC(1/50)[0.43492] CWKAVTWLC(1/50)[0.43425] CPLGFVGDC(1/50)[0.26286] CNTSAYPDC(1/50)[0.22191] 14 Phage display (subtractive panning) 4 PMID:31247207 Plasma from 10 healthy persons Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The absorbance value at 450 nm was determined with a microplate reader. Data (OD450) shown were reproduced from Figure 2D in the reference. In the first round of selection, phage library was selected against the pooled control plasma with albumin/IgG depletion. The second round was negative selection against pooled plasma from 10 Alzheimer's disease (AD) patients (5 male and 5 female) without albumin/IgG depletion. The third and fourth rounds of selection were against the same pooled control plasma without albumin/IgG. We further characterized one AD-specific peptide (AD#1 peptide, CHMRQGMAC) and one control-specific peptide (Con#1 peptide, CDGARHGRC), and evaluated their diagnostic performance in independent validation set (35 AD patients, 45 MCI, 45 controls and 20 PD patients). Our results show that both AD#1 peptide and Con#1 peptide could distinguish AD/MCI patients from controls and combition of these two peptides could greatly improve the diagnostic performance (AUC is above 0.80 in ROC curve alysis). 3493 NHRKVSRHATHF(4)[0.08643] PRHGKKPTNKRK(3)[0.79724] TVDSASLLQSRT(1)[0.08422] WGFHWPVYPPSR(1)[0.07299] ADARPWWKSQGF(1)[0.08063] FPYPMNKQTNGT(1)[0.08574] 6 Phage display (competitive panning) 3 PMID:31823885 Lectin α2,6-sialyllactose, 6'-SL Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance was determined at 415 nm in a microtiter plate reader (Tecan Infinite® M1000 Pro, Tecan, Switzerland). Data (OD415) shown were reproduced from Figure 1 in the reference. Bound phages were eluted using 100 μL of 1 M α2,6-sialyllactose (6′-SL) (Cat# 35890-39-2, Dextra Laboratories, Reading, England, UK) in each round of panning. Mimetic peptide (PRHGKKPTNKRK), reverse peptide (KRKNTPKKGHRP) and scrambled peptide (RPGHKKTPKNKR) were tested for inhibition of 6′-SL binding to the lectin. Indeed, lectin binding to 6′-SL was inhibited by the mimetic peptide , but not by the reverse or scrambled peptides, showing that this peptide mimics 6′-SL. Functiolly, mimetic peptide, but not the reverse or scrambled peptides, increased viability and expression of neural cell adhesion molecule L1 in SK-N-SH human neuroblastoma cells, and promoted survival and neurite outgrowth of cultured mouse cerebellar granule neurons challenged by H2O2-induced oxidative stress. The combined results indicate that the 6′-SL mimetic peptide promotes neurol survival and neuritogenesis, thus raising hopes for the treatment of neurodegenerative diseases. 3494 TMANEAPYPGTQ[1.41642] HWDPPYPGSGTQ[1.32673] TEAPYPGSSVTN[1.20625] SALIDAAYPGTQ[1.14333] SEAPYPGAGVYN[1.1942] TFEAPYPGTMLV[1.12995] STEAPYPGIAVQ[1.31646] EAPYPG 7 Phage display (common panning) 3 PMID:31500272 Anti-σA monoclonal antibody 4E2 σA protein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Reactivity of phage clones to MAb 4E2 was determined by enzyme-linked immunosorbent assay. Data (OD450) were reproduced from Figure 3a in the reference. Purified MAb 4E2 was selected for epitope mapping under three rounds of biopanning with the Ph.D-12 TM Phage Display Peptide Library Kit (New England BioLabs, Cambridge, MA, USA). Sequencing of the phage clones with high OD values revealed the consensus sequence EAPYPG, which was identical to the 56EAPYPG61 (aa 217 to 223) of the DRV σA protein. Residues in epitope 56EAPYPG61 were completely homologous in DRV, GRV, ARV, and TRV. 3495 AFWSVPGTVSMT[1.21845] NWVVGGSVAAVT[1.31435] YNFVMAGLVATT[1.44127] IWVMAGAISLSM[1.07978] YVMAGLILSIPN[1.36512] CFTMASLITITA[1.25935] GAWVVAGLIMTV[1.23397] QWVVAGLVSVLG[1.27345] WV[V/M]AGL[I/V] 8 Phage display (common panning) 3 PMID:31500272 Anti-σA monoclonal antibody 1A7 σA protein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Reactivity of phage clones to MAb 1A7 binding was determined by enzyme-linked immunosorbent assay. Data (OD450) were reproduced from Figure 3b in the reference. Purified MAb 1A7 was selected for epitope mapping under three rounds of biopanning with the Ph.D-12 TM Phage Display Peptide Library Kit (New England BioLabs, Cambridge, MA, USA). Sequencing of the phage clones with high OD values revealed the consensus sequence WVV/MAGLI/V, which was identical to the 341WVVAGLI347 (aa 341–347) sequences of the DRV σA protein. Residues in the 341WVVAGLI347 were homologous in DRV and GRV, but divergent at residues 343V/M or/and 347I/V between DRV/GRV and ARV/TRV. 3496 GDGNSVLKPGNW[1.38809] DRWVARDPASIF[2.9589] 2 Phage display (common panning) 5 PMID:31279172 Neutrophil gelatinase-associated lipocalin Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The ELISA sigl was measured at 405 nm using a microplate spectrophotometer (Multiskan FC, Thermo Scientific, Waltham, MA, USA). Data (OD415) shown were reproduced from Figure S1a in the reference. Three rounds of biopanning were performed against recombint NGAL proteins. After washing, the bound phages were eluted using 100 μL of 0.2 M glycine-HCl (pH 2.2), and the eluent was immediately neutralized with 15 μL of Tris-HCl (pH 9.1) to prevent destruction of the phages.The eluted phages were amplified using E. coli ER2738 to make sufficient copies for subsequent rounds of biopanning. The amplified phages were harvested by Cl/polyethylene glycol precipitation. The neutrophil gelatise-associated lipocalin (NGAL) BP1 (DRWVARDPASIF) was selected as most promising recognition receptor, and its binding affinity was monitored by SWV and EIS. Using EIS, the limit of detection (LOD) was 1.74 ng/mL, while SWV had a LOD of 3.93 ng/mL. The detection performance of the peptide-incorporated sensor was comparable to commercially available ELISA NGAL detection kits. In addition, the validation of the peptide sensor was also confirmed with plasma from patients, and it was observed that the sensitivity of the peptide sensor showed a statistically significant difference. Our results show that the phage and peptide sensor system could detect NGAL with high sensitivity and selectivity, and this suggests its potential use as a biosensing platform for monitoring NGAL in a miniaturized electrochemical biosensor. 3497 HSFKWLDSPRLR(5)[2.580765±0.716041] GAYTSWRTSTNA(3)[1.788102±0.130236] DLVSWAGSGKKH(3)[1.855382±0.037913] VHWDFRQWWQPS(1)[2.125573±0.006749] AHAHTNWTSWWE(1)[2.594787±0.006743] ADWYHWRSHSSS(1)[2.043153±0.005618] IPNYSMQSREYR(1)[2.080793±0.006749] HYRPFTQEHRVT(1)[3.67144] HSFKGWDWPRLR(1)[2.795317±0.006743] GWKSHEPKGHGS(1)[2.694727±0.005618] EHLHASWNFSSG(1)[1.808223±0.006743] YDVPNKSWRTSW(1)[4.17569] 12 Phage display (subtractive panning) 3 PMID:31888393 AdipoR-12C Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The OD405 was measured using a microplate reader (StatFax-2100, Awarness Technology, Fisher Bioblock Scientific, Tournai, Belgium). Data (OD405) shown were reproduced from Figure 1G in the reference. Bovine serum albumin (BSA) immobilized on Dynabeads was used as control protein during the preselection steps of the 3 panning rounds to exclude non-specific phages. The selected peptide P17 (IPNYSMQSREYR) recognizes AdipoR1/R2 expressed by skeletal muscle, liver and pancreatic islets. In HepaRG and C2C12 cells, P17 induced the activation of AMPK (AMPKα-pT172) and the expression of succinate dehydrogenase and glucokinase; no cytotoxic effects were observed on HepaRG cells. In db/db mice, P17 promoted body weight and glycemia stabilization, decreased plasma triglycerides to the range of healthy mice and increased adiponectin (in high fat-fed mice) and insulin (in chow-fed mice) levels. It restored to the range of healthy mice the tissue levels and subcellular distribution of AdipoR1/R2, AMPKα-pT172 and PPARα-pS12. In liver, P17 reduced steatosis and apoptosis. The docking of P17 to AdipoR is reminiscent of the binding mechanism of adiponectin. To conclude, we have developed an AdipoR1/AdipoR2-targeted peptide that modulates adiponectin signalling pathways and has therapeutic relevance for T2D and obesity associated pathologies. 3498 GLKIWSLPPHHG NFMESLPRLGMH FDLNPRSLYGSL NPNGTTGPLDSV SMVYGNRLPSAL NLSNRLNLSPGI KVWTLDFQPPVL SLTVPFLPLYVP QPHKVFFPNLPR GHHSMTPGTAPH FATQQPPTAHIP LLADTTHHRPWT IPWTQHMAMSPM YSDQPTQSSQRP QVYAEFKTSFRS AVLDELRRRIFGA 16 Phage display (subtractive panning) 3 PMID:32840755 Soluble proteins differentially expressed between the control and dimethylhydrazine treated groups Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) For the second and third selection cycles, a pre-selection was carried out first using wells containing immobilized proteins for the control groups. The non-binding peptides (F1) were collected, amplified as previously described, and then used for selection against extracts from the test groups. Two peptides identified by phage display (GLKIWSLPPHHG denoted as peptide-1 and NFMESLPRLGMH peptide-2) were synthesized and exhibited purity higher than 84%. Poly(lactic acid)-block-polyethylene glycol nanospheres were prepared by nanoprecipitation and double emulsion methods in order to load the two peptides. Nanoparticles ranged in size from 114 to 150 nm and peptide encapsulation efficiency varied from 16 to 32%, depending on the methodology. No cytotoxic activity was observed towards Caco-2 tumor cell line, either free or loaded peptides in concentrations up to 3 μM at incubation times of 6 and 24 h, indicating safety as biomarkers. Fluorescein isothiocyanate–labeled peptides allowed evaluating selective interactions with Caco-2 cells, where peptide-1 entrapped in nanospheres showed greater intensity of co-localized cell fluorescence, in comparison to peptide-2. Peptide-1 loaded in nanospheres revealed promising to be investigated in further studies of selectivity with other human colon rectal cells as a potential biomarker. 3499 HSLWMASPMPGY(8/13)[0.12] QIFTSSPMPAMV(5/13)[0.09] 2 3 PMID:32222498 Anti-imidacloprid monoclonal antibody 3D11 Imidacloprid Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The half-maximum inhibition concentration (IC50, ng/mL) was calculated using the calibration curve and shown. For the first round of panning, the bound phages were eluted by adding 100 μL of 1 mg L−1 imidacloprid. To eliminate nonspecifically bound phages, the eluate was transferred into wells that were coated with 5% milk in PBS alone, and then the phages in the supernatants were amplified by infection of Escherichia coli ER2738. The amplified phages were subjected to two further panning procedures, which were carried out using mAb concentrations of 50 and 25 mg L−1, imidacloprid concentrations of 0.1 and 0.01 mg L−1, and PBS containing 0.3% Tween 20 and 0.5% Tween 20, respectively. We isolated two phage-borne peptides that compete with imidacloprid to bind the monoclonal antibody (mAb) 3D11 from phage display peptide libraries. A phage-enzyme-linked immunosorbent assay (P-ELISA) and two phage time-resolved fluoroimmunoassays (P-TRFIAs) for the detection of imidacloprid were developed using the phage-borne peptides as substitutes for chemically synthesized antigens. After systematic optimization, the half-maximum inhibition concentrations (IC50) of the P-ELISA, P-TRFIA-1, and P-TRFIA-2 were 0.067 ng mL−1, 0.085 ng mL−1, and 0.056 ng mL−1, respectively. Based on their IC50 values, the sensitivities of the P-ELISA and P-TRFIAs were more than four times greater than those of previous immunoassays. 3500 CWSYAVHPEC(4)[1.286620±0.009336] CDEYAVHMEC(4)[NA] CEGYAIHPEC(3)[NA] CFWYASHPEC(2)[NA] CIMYAIHPEC(1)[NA] CVLYALHDEC(1)[NA] CQFYVLHPEC(1)[NA] CAFYVMHPEC(1)[NA] CVWYVVHPEC(1)[NA] CILYAVHPEC(1)[NA] CDVYAVHPEC(1)[NA] CGDYAVHAMC(1)[NA] CEFYAVHALC(1)[NA] CVDYAVHVEC(1)[NA] YAVHP 14 Phage display (common panning) 4 PMID:31434316 Anti-LDL (−) monoclonal antibody 1A3H2 Electronegative low-density lipoprotein, LDL (−) Not determined. Not determined. fUSE55-based CX8C phage display library (CX8C) ELISA Absorbance at 450 nm was measured by spectrophotometry using a microplate reader (SynergyTM Mx, Biotek instruments Inc, Winooski, VT, USA). Data (OD450) shown were reproduced from Figure 1B in the reference. To prevent the selection of peptides binding to conserved domains of immunoglobulins,the anti-LDL(-) mAbs 1A3 and 2C7 were immobilized on microtiter plates and incubated in the presence of excess soluble, unrelated mouse immunoglobulin. P1A3 (CWSYAVHPEC) was quickly internalized by bone marrow-derived murine macrophages as shown by confocal microscopy. However, P1A3 did not show pro-inflammatory effects. 3501 YHCQGC(11) DYAVHP(2) YGCQGC(2) YFCOGC(1) YVCQGC(1) YXCQGC(1) YYCQGC(1) LWWYEW(1) LWWDCA(1) YAVHP 9 Phage display (common panning) 4 PMID:31434316 Anti-LDL (−) monoclonal antibody 1A3H2 Electronegative low-density lipoprotein, LDL (−) Not determined. Not determined. fUSE55-based X6 phage display library ELISA To prevent the selection of peptides binding to conserved domains of immunoglobulins,the anti-LDL(-) mAbs 1A3 and 2C7 were immobilized on microtiter plates and incubated in the presence of excess soluble, unrelated mouse immunoglobulin. 3502 CGEFLPORIC(3)[NA] CISTELLPSC(2)[NA] CIDFDLPGRC(2)[NA] CVSSEVLPSC(2)[NA] CMPSVILPSC(1)[0.818810±0.008400] CVSTVMLPSC(1)[NA] CISSVWLPSC(1)[NA] CYSSAMLPSC(1)[NA] CMSSEMLPSC(1)[NA] CVSSDVLPSC(1)[NA] CSSSAILPSC(1)[NA] CHPSAILPSC(1)[NA] CLSSFIIPSC(1)[NA] CSPSWLIPSC(1)[NA] CVPSVLLPSC(1)[NA] CLPSVFLPSC(1)[NA] CVPSYILPSC(1)[NA] CIPSMMLPSC(1)[NA] CLPSMILPSC(1)[NA] CWPSGVLPSC(1)[NA] CFTSVYLPSC(1)[NA] CDTSYLLPSC(1)[NA] CGDVLPSVSC(1)[NA] CGDTLPSVEC(1)[NA] CGSVLPSVVC(1)[NA] CGLYLPSVPC(1)[NA] CLDVFLPGRC(1)[NA] CFDVFLPGRC(1)[NA] CLDFELPGRC(1)[NA] CGESLPSRKC(1)[NA] CGVYLPSRLC(1)[NA] CVGKNFYPSC(1)[NA] CMWIPSGAMC(1)[NA] CLDIIPSGFC(1)[NA] CIQGDVIPSC(1)[NA] VLPS 35 Phage display (common panning) 4 PMID:31434316 Anti-LDL (−) monoclonal antibody 2C7D5F10 Electronegative low-density lipoprotein, LDL (−) Not determined. Not determined. fUSE55-based CX8C phage display library (CX8C) ELISA Absorbance at 450 nm was measured by spectrophotometry using a microplate reader (SynergyTM Mx, Biotek instruments Inc, Winooski, VT, USA). Data (OD450) shown were reproduced from Figure 1B in the reference. To prevent the selection of peptides binding to conserved domains of immunoglobulins,the anti-LDL(-) mAbs 1A3 and 2C7 were immobilized on microtiter plates and incubated in the presence of excess soluble, unrelated mouse immunoglobulin. P2C7 (CMPSVILPSC)) was quickly internalized by bone marrow-derived murine macrophages as shown by confocal microscopy. P2C7 increased the expression of TNFalpha, IL-1beta and iNOS as well as the secretion of TNFalpha, CCL2, and nitric oxide by murine macrophages, similar to the responses induced by LDL (-), although less intense. We identified a mimetic epitope associated with LDL (-), the P2C7 circular peptide, that activates macrophages. Our data suggest that this conformational epitope represents an important danger-associated molecular pattern of LDL (-) that triggers proinflammatory responses. 3503 CEVLPGRC(18) CMWLPGAC(4) GVDDGA(1) GRADIR(1) VLPS 4 Phage display (common panning) 3 PMID:31434316 Anti-LDL (−) monoclonal antibody 2C7D5F10 Electronegative low-density lipoprotein, LDL (−) Not determined. Not determined. ELISA To prevent the selection of peptides binding to conserved domains of immunoglobulins,the anti-LDL(-) mAbs 1A3 and 2C7 were immobilized on microtiter plates and incubated in the presence of excess soluble, unrelated mouse immunoglobulin. 3504 HFPFHHHKLRAH(9/20)[0.699] HPMHMLHKRQHG(5/20)[0.336] HPHWWQVFPKRT(4/20)[0.551] WPNLHHRFHTVR(2/20)[NA] 4 Phage display (common panning) 3 PMID:31733137 19-kDa fragment of Merozoite surface protein 1, MSP-I19 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding affinity of each selected peptide clone to rPkMSP‐119 was validated using phage ELISA. The binding signals for each peptide were estimated using absorbance value at 405 nm towards rPkMSP‐119. Three rounds of biopanning were performed against purified PkMSP‐119 protein. After washing, bound phages were eluted by adding elution buffer 0.2 m Glycine–HCl (pH 2.2) supplemented with 1 mg/ml BSA into well, and the plate was gently rocked for 15 min at room temperature. The eluate was then neutralised with 1 m Tris‐HCl, pH 9.1. The second round of biopanning, 200 μl of 5e11 phage (eluates from first biopanning) was added into the PkMSP‐119 protein coated well and the same process was repeated as first biopanning. Four phage peptide variants, namely Pkd1 (HFPFHHHKLRAH), Pkd2 (HPMHMLHKRQHG), Pkd3 (HPHWWQVFPKRT) and Pkd4 (WPNLHHRFHTVR), that bound to PkMSP‐119 were identified from 20 randomly picked phage isolates in the third round of biopanning. Single Pro (P) and Arg (R) were detected among the three binding peptides (pkd1, pkd2 and pkd3). The sequences of both Pkd1 and Pkd2 consist of a large number of histidine residues. Pkd1 showed positive binding signal with 6.1× vs. BSA control. Docking results showed that Pkd1 and Pkd2 were ideal binding peptides for PkMSP‐119. 3505 CVRARTR(4.1%)[281±92,2.309867±0.004492] CLQKTPKQC(2.7%)[373±114,2.456670±0.006287] CVPRRGR(2.7%)[NA,2.163067±0.005392] CRPVRVR(2.7%)[NA,2.15684] CRIRDPRR(1.2%)[NA,2.28912] CTKRIR(1.2%)[NA,2.28912] CRRAR(1.2%)[NA,2.441107±0.009884] CKLRT(1.2%)[NA,2.750273±0.009884] 8 Phage display (subtractive panning, competitive panning) 3-5 PMID:32278214 Programmed cell death ligand-1 overexpressing HEK 293T cells Not determined. Not determined. Not determined. CX7C T7 phage display library Surface plasmon resonance (SPR) The binding affinity (KD, nM) of PD-L1-binding peptides was measured by the surface plasmon resonance (SPR) method using an SPR instrument (Reichert Technologies, Depew, NY) at 25 °C. The KD value was displayed in the first column of the affinity value. In addition, absorbance at 450 nm was measured by using a microplate reader (Tecan, Zurich, Switzerland). Data (OD450, the second column) shown were reproduced from Figure 1D in the reference. To subtract phage that non-specifically bind HEK 293T cells, the phage library, containing 1e9 plaque-forming units (pfu), was incubated with non-transfected HEK 293T cells at 4 °C for 1 h. Unbound phage were collected from the supernatant and incubated with PD-L1-transfected HEK 293T cells at 4 °C for 1 h. Bound phage was eluted by incubation with the Escherichia coli host BL21 for 10 min at 24 °C or room temperature (RT). The eluates were used for titration of pfu and for amplification in BL21. The biopanning procedure was repeated five times. Two selected peptides, CLQKTPKQC and CVRARTR (PD-L1Pep-1 and PD-L1Pep-2, respectively) that appeared to block PD-L1. PD-L1Pep-1 and PD-L1Pep-2 preferentially bound to high PD-L1-expressing cells over low PD-L1-expressing cells; binding was further enhanced by interferon-γ, an inducer of PD-L1 expression. Binding affinities of PD-L1Pep-1 and PD-L1Pep-2 were approximately 373 and 281 nM, respectively. Cellular binding of the PD-L1-binding peptides was reduced by silencing PD-L1 gene expression or competition with anti-PD-L1 antibody. PD-L1Pep-1 and PD-L1Pep-2 induced the internalization and downregulated cell surface levels of PD-L1. The PD-L1-binding peptides restored cytokine secretion and T-cell proliferation to cells inhibited by co-culture with tumor cells or culture on PD-L1-coated plates. Intravenously injected PD-L1Pep-1 and PD-L1Pep-2 efficiently homed to tumor tissues, inhibited tumor growth, and increased CD8+/FoxP3+ ratio in mice. The PD-L1-binding peptides in combination with doxorubicin or PD-L1-targeted liposomal doxorubicin inhibited tumor growth and increased CD8+/FoxP3+ ratio more efficiently than doxorubicin alone and untargeted liposomal doxorubicin, respectively. These results suggest that PD-L1Pep-1 and PD-L1Pep-2 block PD-L1 and reinvigorate T-cell activity, inhibiting tumor growth by enhancing anti-tumor immunity. 3506 KLWSIPTNFLLP(9/10) NAKYLPTILGRL(1/10) 2 Phage display (competitive panning) 3 PMID:32881706 Major allergen Can f 1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were eluted competitively in 100 μL solution of the respective free target (100 μg/mL Can f1 in TBS) on a rotating shaker at RT for 60 min. Panning procedure was repeated three times. The KLWSIPTNFLLP peptide corresponded to the antigen-binding site of a putative γδT-cell receptor. Additional biochemical investigations confirmed this interaction. 3507 YEHAMYRSAVLL[1.67] HDGTLLPRSSLH[2.16] TIGVVRDVATQL[2.06] ISTVYTGLTEKD[1.31] DSLTMKRLVLPY[1.58] RNVELHDALRRT[1.38] NSSTPMLSDVLR[1.70] NRVSDHLVRAVS[6.71] HAPMPYANWPRG[1.78] HVKAAYPMVTMS[3.42] 10 Phage display (common panning) PMID:31308251 Serum IgG of Systemic lupus erythematosus(SLE) patients Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Optical density was read at 450 nm using a Behring EL311 ELISA microplate reader (Dade Behring Marburg Gmbh, Berlin, Germany). A typical four-parameter logistic nonlinear regression model was used for standard curve fitting for the ELISA, which was further used to estimate sample content (units/ml) from the absorbance measurement data. The concentration of IgG from the samples were measured with the commercial Human IgG ELISA Quantitation kits (eBioscience, Massachusetts) according to the instructions. Besides the serum samples, we also added 1 μg of an anti-6×His antibody (Millipore, California) and 10 μl PBS buffer to a 100 μl immunoprecipitation mix as positive control and negative control, respectively. About 1 × 109 M13-phage particles were added to 100 μl immunoprecipitation mix in a 96-well PCR plate. The plate was then carefully sealed with adhesive optical tape (Applied Biosystems, California) and rotated overnight at 4 °C. 10 μl of protein G Dynabeads (Invitrogen, Massachusetts) were then added to each well. The resealed plate was placed back on a rotator for 4 h at 4 °C. The beads were washed six times in 200 μl wash buffer (150 mm NaCl, 50 mm Tris-HCl, 0.1% NP-40 (pH 7.5)) by pipetting up and down eight times per wash. After removal of the wash buffer, the beads were resuspended in 30 μl ddH2O and heated at 95 °C for 10 min. To discover biomarkers for Systemic lupus erythematosus(SLE),a phage displayed random peptide library (Ph.D. 12) and deep sequencing were applied to screen specific autoantibodies in a total of 100 serum samples from 50 SLE patients and 50 healthy controls. Then, four peptides (YEHAMYRSAVLL, HDGTLLPRSSLH,TIGVVRDVATQL, and ISTVYTGLTEKD) were discovered with high diagnostic power to differentiate Systemic lupus erythematosus(SLE) patients from healthy controls. Among them, two peptides, i.e. YEHAMYRSAVLL and HDGTLLPRSSLH, were confirmed between SLE with other autoimmune patients. 3508 LTPMPNW(3)[1.448491±0.940644] MMSYPKH(2)[2.745750±0.669996] MMTLPNN(1)[3.233583±0.013649] MAREMSY(1)[3.147363±0.008307] VLAPPCW(1)[2.65617] 5 Phage display (common panning) 3 PMID:31255726 ArsS N-terminal (ArsS-N) acid-sensing domain Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA OD450nm was assessed using a microplate reader (T100, Bio-Rad, Hercules, CA, USA). Data (OD450) shown were reproduced from Figure 4 in the reference. To identify peptides that antagonize the acid-sensing domain of H. pylori ArsS. Phage clones were selected by solution-phase panning with protein G magnetic beads (NEB) that exhibited high affinity for purified recombinant Helicobacter pylori ArsS N-terminal acid-sensing protein (P-ArsS-A). The phages were reacted with the magnetic beads at room temperature for 2 h, which were then washed five times with 0.05% PBS with Tween 20 (PBST) for 5 min. To elute the bound phages, 100 μl 0.2M glycine-HCl (pH 2.2) was added for 8 min. Subsequently, 15 μl 1M Tris-HCl (pH 9.1) was used to neutralize the solution. Two additional rounds of panning were performed, with a phage load of 2e11 PFU/ml in each round of panning. Positive monoclonal phages were obtained after a total of three rounds of panning. Eight phage clones that could specifically bind to P-ArsS-A were obtained and five amino acid sequences were identified, including P03 (MMSYPKH) and P06 (LTPMPNW). An in vitro minimum inhibitory concentration (MIC) evaluation showed that P03 and P06 significantly inhibited the growth of H. pylori J99. The MIC of P03 was 8 μM, and the MIC of P06 was >16 μM, indicating that P03 is a stronger inhibitor compared to P06. This was confirmed by colony counting on blood agar plates after P03 and P06 administration. Using homology modeling and molecular docking analysis, it was shown that P03 and P06 could bind to the ArsS N-terminal domain, and there were four shared binding sites: TYR25, ASN39, ARG73, and GLU74. Additionally, one hydrogen bond was found between P03 and ArsS, which is more cohesive than other forms of bonding (van der Waals force, other non-covalent bonds). 3509 DWSSWVYRDPQT(10%) SGVYKVAYDWQH(5%) 2 Phage display (competitive panning) 6 PMID:31902944 Folate receptor alpha Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Bound phages were eluted by addition of 1 mL of folic acid (Acros organics, Fisher Scientific, UK) in PBS (100 µg/mL) by competitive inhibition for 1 h at 4 °C. Two phage peptides(DWSSWVYRDPQT,SGVYKVAYDWQH) showed binding specificity to SKOV3 cells in vitro screened by Ph.D.-12 phage display library. WSSWVYRDPQT peptide identified targets colon cancer cells in vitro with in silico analysis suggesting the peptide targets glypican-3 (a heparin sulphate proteoglycan (HSPG)). 3510 CIGNSNTLC(9%) CTVRTSADC(7%) CTVRTSAEC(4%) 3 Phage display (competitive panning) 6 PMID:31902944 Folate receptor alpha Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Bound phages were eluted by addition of 1 mL of folic acid (Acros organics, Fisher Scientific, UK) in PBS (100 µg/mL) by competitive inhibition for 1 h at 4 °C. Three phage peptides(CIGNSNTLC,CTVRTSADC,CTVRTSAEC) showed binding specificity to SKOV3 cells in vitro screened by Ph.D.-C7C phage display library. Peptides CIGNSNTLC and CTVRTSAEC bound FRα in the context of the phage particle. And synthesised lead peptide, CTVRTSAEC, bound specifically to FRα and could be competitively inhibited with folic acid. To assess the capacity of the elucidated FRα-binding oligopeptides to target OV to FRα, we genetically incorporated the peptides into the HAdV-C5 fiber-knob HI loop including in vectors genetically ablated for hCAR interactions. Unfortunately, the recombinant vectors failed to efficiently target transduction via FRα due to defective intracellular trafficking following entry via FRα, indicating that whilst the peptides identified may have potential for applications for targeted drug delivery, they require additional refinement for targeted virotherapy applications. 3511 SCATPFSPQVCS(78/80)[0.30566] SCRLQVXPGFCS(1/80)[NA] SCGERGXAECCS(1/80)[NA] 3 Phage display (common panning) 4 PMID:31422560 Copper (II) oxide nanoparticles, CuO-NPs Not determined. Not determined. Not determined. T7 SCX8CS phage display library (SCX8CS) ELISA The absorbance of each sample was measured at 450 nm using a microplate reader (Spectra Max Plus 384; Molecular Devices, San Jose, CA, USA). Data (OD450) shown were reproduced from Figure 1b in the reference. Prepared CuO-NPs (500 μg) were mixed with the constructed libraries and incubated in TBS-T containing 0.1% Tween 20. After 1 h at room temperature, CuO-NPs were washed five times with TBS-T. For the proliferation of T7 phages bound to the surface of CuO-NPs, E. coli BLT5403 (Merck Millipore) proliferated to the log phase was mixed with the nanoparticles and incubated at 37 °C. We evaluated the cytotoxicity of copper (II) oxide nanoparticles (CuO-NPs) by coating with a novel cyclic peptide, CuO binding peptide 1 (CuBP1), cyclic-SCATPFSPQVCS, which binds to the surface of CuO-NPs. CuBP1 was found to promote the aggregation of CuO-NPs under mild conditions. The treated CuO-NPs with CuBP1 caused the reduction of the cytotoxicity against Escherichia coli, Lactobacillus helveticus, and five other microorganisms, including bacteria and eukaryotes. Similar effects were also demonstrated against human embryonic kidney (HEK293) cells in vitro. 3512 CRQTKN(9/22)[9.92179] CRGTAEG(6/22)[4.812447±0.023385] CKSRKDGAC(3/22)[3.667137±0.009220] CMPKRPSSC(1/22)[5.607913±0.032665] 4 Phage display (in vivo) 5 PMID:31243044 K-ras(LA2) mutant mouse model of lung cancer Not determined. Not determined. Not determined. T7 CX7C phage display library (CX7C) Binding assay Ex vivo fluorescence intensities were measured using the eXplore Optix and Analysis Workstation software (ART Inc.). Fluorescence intensities were reproduced from Figure 1C in the reference and shown. Data are presented as mean +/- SE. K-ras(LA2) mutant mice bearing lung tumors (18–20 weeks old) were intravenously injected with a solution containing phages at a total of 1e11 pfu. After 15 minutes of circulation, mice were sacrificed and tumor nodules from lungs were excised and weighed. Cell suspension of tumor nodules was prepared in culture media by chopping with a knife, homogenizing using Medimachine (DAKO), and passing through a 70 μm cell strainer. The cell bound phages were eluted by lysing cells with 100 mL of 1% NP-40 for 10 minutes on ice and then by adding BL21 host bacteria. Phages were amplified in the host bacteria and used for next round of selection. Compared with other candidate peptides selected from 5 rounds of phage display, the CRQTKN peptide homed to tumor nodules in the lung of mutant mice at higher levels. Photoacoustic tomography of mutant mice detected lung tumors via tumor homing of the near-infrared fluorescence dye-labeled CRQTKN peptide. Ex vivo photoacoustic images of isolated organs further demonstrated tumor homing of the CRQTKN peptide, whereas minimal accumulation was observed in control organs, such as the liver. Compared with untargeted liposomes and doxorubicin, doxorubicin-loaded liposomes whose surface was modified with the CRQTKN peptide more efficiently delivered doxorubicin and reduced the number or size of tumor lesions in K-ras(LA2) mutant mice. Analysis of hematologic parameters and liver and kidney function showed no significant systemic side effects by the treatments. Affinity-based identification was used to detect TNF receptor superfamily member 19L (TNFRSF19L), which was upregulated in lung tumors of mutant mice, as the receptor for the CRQTKN peptide. In conclusion, these results suggest that the CRQTKN peptide is a promising targeting probe for photoacoustic-guided detection and drug delivery to lung cancer, and acts by binding to TNFRSF19L. 3513 CKSRKDGAC(4/27)[3.667137±0.009220] CMPKRPSSC(3/27)[5.607913±0.032665] CRGTAEG(3/27)[4.812447±0.023385] 3 Phage display (in vivo) 4 PMID:31243044 K-ras(LA2) mutant mouse model of lung cancer Not determined. Not determined. Not determined. T7 CX7C phage display library (CX7C) Binding assay Ex vivo fluorescence intensities were measured using the eXplore Optix and Analysis Workstation software (ART Inc.). Fluorescence intensities were reproduced from Figure 1C in the reference and shown. Data are presented as mean +/- SE. K-ras(LA2) mutant mice bearing lung tumors (18–20 weeks old) were intravenously injected with a solution containing phages at a total of 1e11 pfu. After 15 minutes of circulation, mice were sacrificed and tumor nodules from lungs were excised and weighed. Cell suspension of tumor nodules was prepared in culture media by chopping with a knife, homogenizing using Medimachine (DAKO), and passing through a 70 μm cell strainer. The cell bound phages were eluted by lysing cells with 100 mL of 1% NP-40 for 10 minutes on ice and then by adding BL21 host bacteria. Phages were amplified in the host bacteria and used for next round of selection. Compared with other candidate peptides selected from 4 rounds of phage display, there peptides(CKSRKDGAC,CMPKRPSSC,CRGTAEG) were selected. 3514 WHWRLPS(2)[1.359865±0.157799] NPMHIYDTLPAR(2)[0.400335±0.030196] 2 Phage display (common panning) 3-5 PMID:32717427 Multiple Sclerosis recombinant antibodie A1, MS-A1 (ST-176) Not determined. Not determined. Not determined. Ph.D.-12 and Ph.D.-7 phage library pool ELISA Absorbance at 405 nm was determined spectrophotometrically with a Microplate Reader (Bio-Rad, Hercules, CA). Data (OD405) shown were reproduced from Figure 1A in the reference. The Phage Display Peptide Libraries (12mer + 7mer) (New England BioLabs, Beverly, MA) were used for affinity selection of specific peptides under five rounds of biopannning for the MS brain rAbs. We identified 4 high affinity phage peptides from which 2 peptides (NPMHIYDTLPAR,WHWRLPS) are unique. Database searches revealed that peptide A1-A1(NPMHIYDTLPAR) shared sequence homologies with TRPM8 channel-associated factor 2 and ORF4 polyprotein [Nora virus]. And peptide A1-C3(WHWRLPS) shared sequence homologies with Tyrosine-protein kinase-defective receptor EPH-6, Sodium leak channel non-selective protein and Major structural protein VP1 [Mesocricetus auratus polyomavirus 1]. 3515 AHIPYQGRDTSQ(4) 1 Phage display (common panning) 3-5 PMID:32717427 Multiple Sclerosis recombinant antibodie B1, MS-B1 (G2-160) Not determined. Not determined. Not determined. Ph.D.-12 and Ph.D.-7 phage library pool ELISA The Phage Display Peptide Libraries (12mer + 7mer) (New England BioLabs, Beverly, MA) were used for affinity selection of specific peptides under five rounds of biopannning for the MS brain rAbs. We identified 4 high affinity phage peptides from which 1 peptides(AHIPYQGRDTSQ) are unique. Database searches revealed that peptide B1-A3(AHIPYQGRDTSQ) shared sequence homologies with Cytospin-A and Epstein-Barr nuclear antigen 4. 3516 NISAWFQPHQAL(13)[0.270370±0.088490] WQWGPYMVGAGV(4)[NA] 2 Phage display (common panning) 3-5 PMID:32717427 Multiple Sclerosis recombinant antibodie A2, MS-A2 (ST-196) Not determined. Not determined. Not determined. Ph.D.-12 and Ph.D.-7 phage library pool ELISA Absorbance at 405 nm was determined spectrophotometrically with a Microplate Reader (Bio-Rad, Hercules, CA). Data (OD405) shown were reproduced from Figure 1B in the reference. The Phage Display Peptide Libraries (12mer + 7mer) (New England BioLabs, Beverly, MA) were used for affinity selection of specific peptides under five rounds of biopannning for the MS brain rAbs. We identified 17 high affinity phage peptides from which 2 peptides(NISAWFQPHQAL,WQWGPYMVGAGV) are unique. Database searches revealed that peptide A2-D2(WQWGPYMVGAGV) shared sequence homologies with G-protein coupled receptor 161,Myomesin-3 and RNA-directed RNA polymerase L [Oliveros mammarenavirus]. And peptide A2-F4(NISAWFQPHQAL) shared sequence homologies with Dual specificity protein phosphatase 2,Ubiquitin-like modifier-activating enzyme Epstein-Barr nuclear antigen 6 and Capsid assembly protein [Human herpesvirus 4 strain B95-8]. 3517 SCSSLTTLRPCG(8/20)[1.71407] SQRKLAAKLTSK(6/20)[1.939233±0.016779] VILTGPEAEYFW(3/20)[0.659633±0.004151] HAMSPVFLSKYA(1/20)[NA] QVNGLGERSQQM(1/20)[NA] DYHDPSLPTLRK(1/20)[NA] 6 Phage display (common panning) 4 PMID:31678742 P. aeruginosa H103 (PAO1 wild-type prototroph) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Azino-bis (3-ethylbenzothiazole sulfonic acid) diammonium salt (ABTS) peroxidase substrate (Sigma-Aldrich) was used to detect peptide binding and the colour development at 415 nm was recorded by using a Model 680 microplate reader (Bio-Rad, Hercules, CA, USA). Data (OD415) shown were reproduced from Figure 1A in the reference. Four rounds of biopanning were performed against whole cells of P. aeruginosa H103. After washing, the bound phages were eluted by the addition of 100 μL elution buffer (0.2 M glycine-HCl, pH 2.2, 1 mg/mL BSA) at room temperature for 10 min.After neutralization with 1 M Tris-HCl, pH 9.1, the eluted phage was amplified in E. coli ER2738 by an infection method. The amplified phages were titred and used for the second round of biopanning. After four rounds of biopanning, the eluted phage was used to prepare phage stocks to isolated phage genomic DNA for nucleotide sequencing. The DNA sequences were translated into amino acids by using the Snapgene viewer software. Eight clones encoded the sequence SCSSLTTLRPCG, referred to as PA1, six clones encoded the sequence SQRKLAAKLTSK, referred to as PA2, and three clones encoded the sequence VILTGPEAEYFW, referred to as PA3. The remaining three clones (PA4, 5, and 6) encoded unique peptide sequences. The targeting peptide (PA2) that binds specifically to OprF porin on P. aeruginosa and a hybrid peptide was constructed by addition of the targeting peptide to GNU7, a potent antimicrobial peptide. The resulting hybrid peptide PA2-GNU7 exhibited potent antimicrobial activity against P. aeruginosa without causing host toxicity. Confocal laser scanning microscopy analysis and time-kill experiments demonstrated that PA2-GNU7 exhibited a high degree of specificity for P. aeruginosa, and rapidly and selectively killed P. aeruginosa cells in mixed cultures. In addition, in vivo treatment efficacy of PA2-GNU7 was significantly greater than that of conventional antibiotics in a mouse model of MDR P. aeruginosa infection. Taken together, the data suggest that PA2-GNU7 may be a promising template for further development as a novel anti-MDR P. aeruginosa therapeutic agent. 3518 GSAPLLTVDTSK(22)[12.45 ± 7.40] RVAPDFSCAYPY(18)[12.62 ± 11.01] SGVYKVAYDWQH(13)[35.80 ± 18.22] QSHDQNNYNQRS(7)[NA] 4 Phage display (subtractive panning) 4 PMID:32828793 Amyloid beta 42, Aβ42 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance was measured at 450 nm (OD450) using a microplate reader (Molecular Devices). The Kd value (pM) was determined and shown. Negative selection was performed for reducing non-specific phages between the third and forth round of panning, adding the amplified phage to non-target coated plate for 1 h. Non-bound phages in the supernatant were used for the forth round of biopanning. The synthesized peptides were confirmed to have excellent binding affinity to Aβ aggregates, by immunohistochemical staining and western blotting using the brains of 3X transgenic (Tg) AD mice at different stages (5–7, 12–17 months old) of AD severity. In the present study, it was confirmed that newly developed amyloid-binding peptides could be used as novel probes for the detection of Aβ aggregates, which can be used for clinical diagnosis of AD in the future. 3519 SPAQHTHERAHT(21)[13.07 ± 5.88] SPHLHTSSPWER(17)[217.97 ± 27.01] MKAHHSQLYPRH(12)[1548.22 ± 835.51] NYPHPHLSNYHP(8)[637.02 ± 815.98] NAPKHAHRLPVH(2)[1282.66 ± 269.73] 5 Phage display (subtractive panning) 4 PMID:32828793 Amyloid beta 42, Aβ42 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance was measured at 450 nm (OD450) using a microplate reader (Molecular Devices). The Kd value (pM) was determined and shown. Two rounds of negative selection were performed for reducing non-specific phages, by adding the amplified phage to non-target-coated resin for 1 h. One was performed after the second round of positive selection, the other after the third round of positive selection. After centrifugation, non-bound phages in the supernatant were used for the following cycles. The synthesized peptides were confirmed to have excellent binding affinity to Aβ aggregates, by immunohistochemical staining and western blotting using the brains of 3X transgenic (Tg) AD mice at different stages (5–7, 12–17 months old) of AD severity. In the present study, it was confirmed that newly developed amyloid-binding peptides could be used as novel probes for the detection of Aβ aggregates, which can be used for clinical diagnosis of AD in the future. 3520 MKAHHSQLYPRH[14.269347±0.042793] HRPYLQSHHAKM[16.550423±0.042787] SGHFFMKDHWDV[2.154010±0.017321] VDWHDKMFFHGS[2.64163] HAPDTIKRSLAM[17.884483±0.042793] DKDVTHFLERTR[17.678607±0.042787] NHHMMPAWNVKH[10.816170±0.038036] NLGAEPGTPYLV[15.191657±0.042787] YQPAREHRVPAG[21.867440±0.038036] TNNNTPSQMGLS[13.025873±0.042787] 10 Phage display (common panning) 5 PMID:32375468 Ni–Ti Stents Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay The stent or coil was observed by an upright fluorescence microscope (BX53-44-FL-1, Olympus, Tokyo, Japan).Fluorescence intensity of Ni−Ti stents surfaces after exposure to FITC-labeled peptides was determined. Fluorescence intensities were reproduced from Figure 2B in the reference and shown. To wash out nonbinding phages, the Ni–Ti stents were rinsed by PBS containing Tween 20 (0.1%) once and then by PBS four times at room temperature. The phages bound to the surface of the stent and coil were eluted with 37.5 μg/mL glycine/HCl solution (pH 2.2) containing 1 mg/mL BSA and 0.1 mg/mL phenol red for 15 min at room temperature. The eluted phage solution was neutralized with Tris-HCl (pH 9.1). Then, E. coli cells were infected with the eluted phages and cultured for the phage amplification. These amplified phages were used for the next round of panning. After the final panning at the fifth round, E. coli cells infected with the obtained phages were plated and incubated on LB-agar plates containing IPTG and X-gal (overnight at 37 °C). Phage display method was used to identify peptide ligands that had high affinity for the metallic surface of Ni–Ti stents. The binding assay using fluorescence labeling revealed that several synthetic peptides could bind onto those surfaces. 3521 AHNHTPIKQKYL[7.41469] LTPHKHHKHLHA[23.9808] SAVQWFELNTHA[5.38701] 3 Phage display (common panning) 5 PMID:32375468 Pt–W Coils Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay The stent or coil was observed by an upright fluorescence microscope (BX53-44-FL-1, Olympus, Tokyo, Japan).Fluorescence intensity of Pt−W coils surfaces after exposure to FITC-labeled peptides was determined. Fluorescence intensities were reproduced from Figure 3B in the reference and shown. To wash out nonbinding phages, the Pt–W coils were rinsed by PBS containing Tween 20 (0.1%) once and then by PBS four times at room temperature. The phages bound to the surface of the stent and coil were eluted with 37.5 μg/mL glycine/HCl solution (pH 2.2) containing 1 mg/mL BSA and 0.1 mg/mL phenol red for 15 min at room temperature. The eluted phage solution was neutralized with Tris-HCl (pH 9.1). Then, E. coli cells were infected with the eluted phages and cultured for the phage amplification. These amplified phages were used for the next round of panning. After the final panning at the fifth round, E. coli cells infected with the obtained phages were plated and incubated on LB-agar plates containing IPTG and X-gal (overnight at 37 °C). Phage display method was used to identify peptide ligands that had high affinity for the metallic surface of Pt–W coils. The binding assay using fluorescence labeling revealed that several synthetic peptides could bind onto those surfaces. 3522 HSQSAEHHTSWP[12.77396] TLLPKNFGHRPL[118.495593±0.214277] TFLNSVPTYSYW[53.734663±0.242966] TSCFGDSRCQPP[23.902547±0.242966] LTKTNLDPMEFK[36.52743] TICWTYPKCTED[16.42114] TSNLWRYDRLTM[65.33084] STVSSLARILTG[61.356347±0.242966] 8 Phage display (common panning) 5 PMID:32375468 Co–Cr Stents Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay The stent or coil was observed by an upright fluorescence microscope (BX53-44-FL-1, Olympus, Tokyo, Japan). Fluorescence intensity of Co−Cr stents after exposure to the FITC-peptide was determined. Fluorescence intensities were reproduced from Figure 4B in the reference and shown. To wash out nonbinding phages, the Co–Cr stents were rinsed by PBS containing Tween 20 (0.1%) once and then by PBS four times at room temperature. The phages bound to the surface of the stent and coil were eluted with 37.5 μg/mL glycine/HCl solution (pH 2.2) containing 1 mg/mL BSA and 0.1 mg/mL phenol red for 15 min at room temperature. The eluted phage solution was neutralized with Tris-HCl (pH 9.1). Then, E. coli cells were infected with the eluted phages and cultured for the phage amplification. These amplified phages were used for the next round of panning. After the final panning at the fifth round, E. coli cells infected with the obtained phages were plated and incubated on LB-agar plates containing IPTG and X-gal (overnight at 37 °C). Phage display method was used to identify peptide ligands that had high affinity for the metallic surface of Co–Cr stents. The binding assay using fluorescence labeling revealed that several synthetic peptides could bind onto those surfaces. 3523 WALRVKAG[0.85168, 8.18e-4] SWGKMAKG[0.84287, NA] PAMARKMG[1.10182, NA] 3 Phage display (subtractive panning) 5 PMID:30903640 Interleukin-13 receptor subunit alpha-2 Interleukin-13, IL-13 3LB6, Not determined. T7 X7 phage display library (X7) ELISA,Surface plasmon resonance (SPR) The binding affinity of IL13Rα2 protein binding peptides was measured by ELISA, absorbance at 450 nm was measured by using a microplate reader (GE Healthcare Biosciences). Data (OD450, the first column) shown were reproduced from Supplementary Figure S1a in the reference. In addition, the surface plasmon resonance (SPR) method using an SPR instrument (Reichert Technologies, Depew, NY) at 25 °C. The Kd (M) value was displayed in the second column of the affinity value. For biopanning against IL‐13Rα2, a solution of the constructed T7 phage display library (3.6 × 109 pfu) was added to a well coated with BSA and incubated for 1 hr at room temperature. After incubation, the unbound T7 phages were removed and added to a well coated with IL‐13Rα1/Fc protein, incubated for 1 hr at room temperature, and the unbound T7 phages were removed and added to a well coated with IL‐13Rα2/Fc protein. The well was washed at least 10 times with PBS containing 0.2% Tween‐20 (PBST) after incubation for 10 min at room temperature, and E. coli BL21 cells were added for culturing and phage propagation. The A2b11 peptide (WALRVKAG), which was one of the positive clones, was shown to bind to IL‐13Rα2 protein by Biacore analysis and a binding assay using glioblastoma (GB) cell lines. This peptide was linked with a lytic peptide containing a linker sequence to form the IL‐13Rα2–lytic hybrid peptide. The IL‐13Rα2–lytic hybrid peptide showed cytotoxic activity against GB cell lines in vitro. The IL‐13Rα2–lytic hybrid peptide also affected Akt and Erk1/2 activation following treatment with interleukin‐13 and induced rapid ATP dynamics in GB cells. Anti‐tumor activity of the IL‐13Rα2–lytic hybrid peptide was observed in vivo after intratumoral injection in a mouse xenograft model of human GB cells. These results suggest that the IL‐13Rα2–lytic hybrid peptide might be a potent therapeutic option for patients with GB. 3524 ANLNLWTDYIRW[2.61538] 1 Phage display (common panning) 3 PMID:32454443 Human colon cancer cell line SW480 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding ability of the selected phage clones was evaluated on colon cancer cells (SW480, HT29, HCT116, LoVo, and DLD1) as well as on a normal cell line (CCD-18co) by cell phage ELISA. Absorbance was measured at 450 nm (OD450) using a plate reader. The binding affinities (relative OD values) showed more than two- to five-fold higher selectivity for colorectal cells than the negative controls. The binding affinity (relative fold) was reproduced from the Supplementary Figure 1 and shown. The Ph.D.-12 Phage Display Peptide Library(New England BioLabs, Beverly, MA) were used for affinity selection of specific peptides under three rounds of biopannning for the human colon cancer cells SW480. The phage clone with the highest optical density (450 nm) compared to the negative control cells was selected. This peptide(ANLNLWTDYIRW) probe maintained binding affinity even after serum incubation. For therapeutic applications, this peptide probe was conjugated to hematoporphyrin, a photosensitizer, which showed a significantly enhanced cellular uptake and high photodynamic effect to kill tumor cells. As another application, we made a nanoparticle modified from the peptide probe. It efficiently delivered SN-38, an anticancer drug, into tumor cells, and its tumor-targeting ability was observed in vivo after intravenous injection to the same xenograft model. 3525 ANLNLWTDYIRW[2.25641] 1 Phage display (common panning) 3 PMID:32454443 Human colon cancer cell line HCT116 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding ability of the selected phage clones was evaluated on colon cancer cells (SW480, HT29, HCT116, LoVo, and DLD1) as well as on a normal cell line (CCD-18co) by cell phage ELISA. Absorbance was measured at 450 nm (OD450) using a plate reader. The binding affinities (relative OD values) showed more than two- to five-fold higher selectivity for colorectal cells than the negative controls. The binding affinity (relative fold) was reproduced from the Supplementary Figure 1 and shown. The Ph.D.-12 Phage Display Peptide Library(New England BioLabs, Beverly, MA) were used for affinity selection of specific peptides under three rounds of biopannning for the human colon cancer cells HCT116. The phage clone with the highest optical density (450 nm) compared to the negative control cells was selected. This peptide(ANLNLWTDYIRW) probe maintained binding affinity even after serum incubation. For therapeutic applications, this peptide probe was conjugated to hematoporphyrin, a photosensitizer, which showed a significantly enhanced cellular uptake and high photodynamic effect to kill tumor cells. As another application, we made a nanoparticle modified from the peptide probe. It efficiently delivered SN-38, an anticancer drug, into tumor cells, and its tumor-targeting ability was observed in vivo after intravenous injection to the same xenograft model. 3526 ANLNLWTDYIRW[2.97436] 1 Phage display (common panning) 3 PMID:32454443 Human colon cancer cell line HT29 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding ability of the selected phage clones was evaluated on colon cancer cells (SW480, HT29, HCT116, LoVo, and DLD1) as well as on a normal cell line (CCD-18co) by cell phage ELISA. Absorbance was measured at 450 nm (OD450) using a plate reader. The binding affinities (relative OD values) showed more than two- to five-fold higher selectivity for colorectal cells than the negative controls. The binding affinity (relative fold) was reproduced from the Supplementary Figure 1 and shown. The Ph.D.-12 Phage Display Peptide Library(New England BioLabs, Beverly, MA) were used for affinity selection of specific peptides under three rounds of biopannning for the human colon cancer cells HT29. The phage clone with the highest optical density (450 nm) compared to the negative control cells was selected. This peptide(ANLNLWTDYIRW) probe maintained binding affinity even after serum incubation. For therapeutic applications, this peptide probe was conjugated to hematoporphyrin, a photosensitizer, which showed a significantly enhanced cellular uptake and high photodynamic effect to kill tumor cells. As another application, we made a nanoparticle modified from the peptide probe. It efficiently delivered SN-38, an anticancer drug, into tumor cells, and its tumor-targeting ability was observed in vivo after intravenous injection to the same xenograft model. 3527 ANLNLWTDYIRW[2.07692] 1 Phage display (common panning) 3 PMID:32454443 Human colon cancer cell line LoVo Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding ability of the selected phage clones was evaluated on colon cancer cells (SW480, HT29, HCT116, LoVo, and DLD1) as well as on a normal cell line (CCD-18co) by cell phage ELISA. Absorbance was measured at 450 nm (OD450) using a plate reader. The binding affinities (relative OD values) showed more than two- to five-fold higher selectivity for colorectal cells than the negative controls. The binding affinity (relative fold) was reproduced from the Supplementary Figure 1 and shown. The Ph.D.-12 Phage Display Peptide Library(New England BioLabs, Beverly, MA) were used for affinity selection of specific peptides under three rounds of biopannning for the human colon cancer cells LoVo. The phage clone with the highest optical density (450 nm) compared to the negative control cells was selected. This peptide(ANLNLWTDYIRW) probe maintained binding affinity even after serum incubation. For therapeutic applications, this peptide probe was conjugated to hematoporphyrin, a photosensitizer, which showed a significantly enhanced cellular uptake and high photodynamic effect to kill tumor cells. As another application, we made a nanoparticle modified from the peptide probe. It efficiently delivered SN-38, an anticancer drug, into tumor cells, and its tumor-targeting ability was observed in vivo after intravenous injection to the same xenograft model. 3528 CGGQDLKSC CSNLTSP*C CPSSSREKC 3 Phage display (common panning) 3 PMID:32243979 Cystic fibrosis mucus model Not determined. Not determined. Not determined. CX7C T7 phage display library (CX7C) For the first round of selection, an initial phage amount of 4.2e9 phage plaque forming units (pfu) was added on top of the mucus layer in the donor compartment and incubated for 1 h at room temperature (25 °C). At 5, 10, 15, 30, and 45 min, 10 μL aliquots of the eluates were taken for further quantification. After 1 h, the entire eluate was collected from the basolateral side and titered using standard double layer plaque assay to quantify phage concentration. The eluted phage library was amplified in BL21 E. coli as described before, which was then quantified by plaque assay prior to the next round of selection. Two subsequent rounds of iterative selection were performed with initial amounts of 5.2e6 pfu and 4.2e9 pfu, respectively. We used combinatorial peptide-presenting phage libraries and next-generation sequencing (NGS) to identify hydrophilic, net-neutral charged peptide coatings that enable penetration through human cystic fibrosis (CF) mucus ex vivo with ~600-fold better penetration than control, improve uptake into lung epithelial cells compared to uncoated or PEGylated-nanoparticles, and exhibit enhanced uniform distribution and retention in the mouse lung airways. These peptide coatings address multiple delivery barriers and effectively serve as excellent alternatives to standard PEG surface chemistries to achieve mucus penetration and address some of the challenges encountered using these chemistries. This biomolecule-based strategy can address multiple delivery barriers and hold promise to advance efficacy of therapeutics for diseases like CF. 3529 EGGDSRH[0.137913±0.000046] RLRAIFS[0.134717±0.000040] VSRVAPC[0.129523±0.000046] WLEVYLG[0.138723±0.000046] 4 Phage display (common panning) 3 PMID:31344770 Pancreatic cancer cell line MIA PaCa-2 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Immunocytochemistry (ICC), enzyme-linked immunosorbent assay (ELISA), and fluorescence- activated cell sorter (FACS) were performed to examine the specific binding. Absorbance was determined at 450 nm. OD450 was reproduced from Fig. 1 in the reference. M5(WLEVYLG) bound to Ce6 showed a significantly lower cell survival rate than that of Ce6 alone in photodynamic therapy, which was observed consistently as a change in the tumor size and fluorescence intensity in MIA PaCa-2 cell-implanted animal models. The noble peptide, M5, binds specifically to the pancreatic cancer cell line, MIA PaCa-2. The M5 peptide has potential use in future optical diagnostic and therapeutic purposes. 3530 SGVYKVAYDWQH(5)[NA] SWFSDWDLELHA(5)[56.40442] SQDIRTWNGTRS(3)[NA] ETWLRWSEKYPT(2)[66.06515] 4 Phage display (common panning) 3 PMID:32892011 N-propyl paraben (PP) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The binding affinity and selectivity test of the selected peptide that has binding affinity to target was carried out using Dynabeads® M-270 Amine. Data (ppm) shown were reproduced from Figure 1b in the reference. The Ph.D.-12 Phage Display Peptide Library(New England BioLabs,Beverly, MA, USA) were used for affinity selection of specific peptides under three rounds of biopannning for the n-propyl paraben (PP)(≥99.0%) . We successfully screened two peptides specific to PP, namely PP3; the results showed that the PP concentration reduction in PP3 was the highest at 39%, and the specificity was measured by the capture rate between target and control. 3531 VHWDFRQWWQPS(7)[NA] DSQFNKYSIATV(3)[62.18905] SGVYKVAYDWQH(2)[NA] SQDIRTWNGTRS(1)[NA] 4 Phage display (common panning) 3 PMID:32892011 Methyl paraben (MP) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The binding affinity and selectivity test of the selected peptide that has binding affinity to target was carried out using Dynabeads® M-270 Amine. Data (ppm) shown were reproduced from Figure 1a in the reference. The Ph.D.-12 Phage Display Peptide Library(New England BioLabs,Beverly, MA, USA) were used for affinity selection of specific peptides under three rounds of biopannning for the Methyl paraben (MP) (purity: ≥ 99.0%) . We successfully screened two peptides specific to MP namely, MP4; the results showed that the MP concentration reduction in MP4 was the highest at 44%, and the specificity was measured by the capture rate between target and control. 3532 VHWDRQWWQPS(12) SGVYKVAYDWQH(1) SQDIRTWNGTRS(1) STPIFAEATARS(1) 4 Phage display (common panning) 3 PMID:32892011 Petri dish (polystyrene, PS) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The Ph.D.-12 Phage Display Peptide Library (New England BioLabs,Beverly, MA, USA) were used for affinity selection of specific peptides under three rounds of biopannning for the petri dish. 3533 VHWDFRQWWQPS[0.40 ± 0.03] WHITPWAWWRPM[0.32 ± 0.02] LMTTAGTAFSLA[0.27 ± 0.03] AHKSKLHQHVMF[0.56 ± 0.06] KPHPKVPLEHWR[0.30 ± 0.03] DWSSWVYRDPQP[0.19 ± 0.04] 6 Phage display (common panning) 5 PMID:32166528 Trans-2-nonenal Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance was measured at 405–415 nm. The OD405 was reproduced from Figure 2 in the reference and shown. The Ph.D.-12 Phage Display Peptide Library(New England BioLabs,Beverly, MA, USA) were used for affinity selection of specific peptides under three rounds of biopannning for the trans-2-nonenal. The phage with sequence of AHKSKLHQHVMFGGG (called as P4) in the end of tail, has shown the highest response. To explore a role of the peptide selected in sequence analysis and ELISA assay, the peptide was connected to magnetic beads. The peptide-coated beads were treated within trans-2-nonenal: treatment of P4 peptide shows significant decrease of trans-2-nonenal compared to negative peptide. Based on our results, it is suggested that the peptide, which is selected by phage display, could be used for the removal of trans-2-nonenal and odor associated with aging. 3534 VHWDFRQWWQPS WHITPWAWWRPM 2 Phage display (common panning) 5 PMID:32166528 Petri dish (polystyrene, PS) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3535 LMTTAGTAFSLA 2 Phage display (common panning) 5 PMID:32166528 Glass dish Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3536 QSHDQNNYNQRS(33/69)[0.86 ± 0.62] DSFNGRYEHVPH(12/69)[0.3 ± 0.13] QTNILTRIHRNR(11/69)[0.95 ± 0.41] KPMRRHRSLHRR(7/69)[3.6 ± 1.3] RKPPRNTMQIRI(2/69)[6.3 ± 2.1] RPRTMPMPTSMR(2/69)[2.8 ± 0.95] RTPLRPIRSRKP(1/69)[8.4 ± 3.8] RINRTRRIRPTP(1/69)[8.39 ± 3.7] 8 Phage display (competitive panning) 5 PMID:32619663 Anthrax toxin receptor 1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured using a SpectroMax M2 Multi-Mode Microplate Reader (Molecular Devices) at 450 nm. The value (OD450) shown in Table S2 in the reference was obtained in a binding saturation curve fitting from three independent experiments. The Ph.D.-12 Phage Display Peptide Library (New England BioLabs) were used for affinity selection of specific peptides under five rounds of biopannning for the purified recombinant ATR/TEM8 VWA (rATR/TEM8 VWA). In the first round, 0.1% Tween-20 was used, and In subsequent biopanning (from the 2nd through 5th rounds), the concentration of detergent (Tween-20) or salt (NaCl) in the wash buffer was increased, creating progressively more harsh conditions as follows: 0.3% (2nd round) and 0.5% (3rd to 5th round) Tween-20, and 500 mM NaCl (4th and 5th round). After the final round of biopanning, the protein-bound phages were eluted by the addition of a 10 molar excess of rATR/TEM8 VWA. We selected ATR-binding peptides by using a phage display: among these, we identified two novel peptides(QSHDQNNYNQRS,DSFNGRYEHVPH) binding to the ATR with high affinity and specificity, and that neutralized anthrax toxicity in cells. Furthermore, to enhance the functional efficiency of the probes, the peptides were modified and conjugated to three polyvalent probe backbones: a 17 amino-acid-based cyclic form penta-unit, poly-D-lysine (PDL), or the M13 bacteriophage. One of the functionally modified polyvalent peptide probes, the penta-unit-conjugated probe (PUCP) produced the most potent neutralization of anthrax toxin, with half-maximal inhibitory concentration (IC50) of 20 nM. The PUCP disrupted anthrax toxin binding to its receptor and reduced endocytosis of anthrax toxin. This peptidebased approach may, therefore, represent a promising strategy to combat anthrax toxicosis and other bacterial diseases and may be efficient for disease treatment. 3537 MISTSRK9(40)[0.37345] QKRNTIT(2)[0.20761] 2 Phage display (subtractive panning) 3 PMID:31092638 Patients' sera pool containing Alternaria specific IgE (s-IgE) Anti-human IgE mAbs Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Absorbance was measured at 450 nm (OD450). OD450 was reproduced from Fig.3a in the reference. Phages from the library of phage (Ph.D.-7 phage Display Peptide Library Kit; New England BioLabs) were screened by non-Alternaria s-IgE (plate A) for three-round selections (each time for a new plate A). The nonbinding phages were harvested and then screened by Alternaria s-IgE (plate B) three times (each time for a new plate B). The bound phages (targeted phages) were washed off from plate B. Third, the targeted phages in a monoclonal colony were picked and amplified respectively, and then the amplified phages were purified by polyethylene glycol/NaCl-precipitation and titrated. Two mimotopes (MISTSRK and QKRNTIT) presented high binding ability with the sera of the Alternaria-allergic patients and mice and, therefore, were selected for immunotherapy in the mouse model. Compared with irrelevant phage control, model, and natural extract immunotherapy group, mimotope immunotherapy group significantly reduced serum IgE levels, inflammatory cells infiltration in the lung tissue, and IL-4 levels in bronchoalveolar lavage fluid, whereas serum IgG1 and IFN-γ levels in bronchoalveolar lavage fluid were increased. The results indicate that B cell mimotopes of Alternaria alleviates allergic response in a mouse model and have potential as novel therapeutic agents for IgE-mediated Alternaria-allergic diseases. 3538 GYFDVVLGGFGP(3)[0.49593] SLHTGATNLYLH(1)[0.62228] FIRPNDWGFGPW(1)[0.54192] HVLNSTVWNTRI(1)[0.57257] HSAQASITIKMA(1)[0.631180±0.001074] SYFDALERMLPG(1)[0.72191] DX(1,4)GFGP 6 Phage display (competitive panning) 4 PMID:31452334 Anti-fibronectin-binding protein A (FnBPA) N-terminal A domain (FnBPA-A) polyclonal antibody Fibronectin-binding protein A Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding ability of the selected six mimotope peptides synthesized in vitro reacted with anti-FnBPA-A antibodies was measured using a microplate reader (Model 450; BioRad Laboratories) at 450nm. OD450 was reproduced from Fig.4 in the reference. To obtain phages binding to anti-rFnBPA-A antibodies, a random Ph.D.-12 phage display peptide library (New England Biolabs) was screened with purified anti-FnBPA-A antibodies according to the manufacturer's instructions. For each round of biopanning, phages (1.5e12 PFU/mL diluted with pure normal rabbit IgG) were applied to a 96-well plate precoated with anti-rFnBPA-A antibodies (10 μg/well). Eight anti-rFnBPA-A antibody-binding phage clones were selected for sequencing, and six different 12-mer peptides were displayed by these phages. Although these displayed peptides shared no more than three consecutive amino acid residues identical to the sequence of FnBPA-A, they could be recognized by the FnBPA-A-specific antibodies in vitro and could induce specific antibodies against FnBPA-A in vivo, suggesting that these displayed peptides were mimotopes of FnBPA-A. Finally, the protective efficiencies of these mimotopes were investigated by mouse vaccination and challenge experiments. Compared with that of control group mice, the relative percent survival of mice immunized with phage clones displaying a mimotope was 13.33% (C2: SLHTGATNLYLH or C15: HVLNSTVWNTRI), 0% (C8: GYFDVVLGGFGP), 6.67% (C10: FIRPNDWGFGPW), 26.67% (C19: HSAQASITIKMA or 1:2 mixture of C23: SYFDALERMLPG and C19), 53.33% (C23), 33.33% (1:1 mixture of C23 and C19), and 66.67% (2:1 mixture of C23 and C19). Overall, five peptides mimicking FnBPA-A protein epitopes were obtained, and a partially protective immunity against S. aureus infection could be stimulated by these mimotope peptides in mice. 3539 PSPHRQRQHILR(25/50) QTIRIIIRRSRT(6/60) SLHMRHKRKPRR(4/50) SSRSMQRTLIIS(2/50) IRSIRMRRILIL(2/50) KTSMRPLILIHI(2/50) 6 Phage display (subtractive panning) 4 PMID:32417652 Brain tumor initiating cells (BTICs) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A BTIC-specific phage library was generated by subtracting against U87MG cells, U251 N cells, and human normal fetal astrocytes to deplete the library of background phage and common binders, and a mixture of patient-derived BTICs were used for selection of candidate phage. A combinatorial phage display library was biopanned against glioblastoma cell model systems that accurately recapitulate the intra- and inter-tumor heterogeneity and infiltrative nature of the disease. Candidate peptides were screened for specificity and ability to target glioblastoma cells in vivo. Six unique 12 amino acid (12mer) peptide sequences were identified more than once, and four were retained (PSPHRQRQHILR,QTIRIIIRRSRT,SLHMRHKRKPRR,SSRSMQRTLIIS) based on back screening of the synthetically made corresponding peptides against a non-invasive tumor cell line used for subtraction. 3540 SVSVGMKPSPRP(21/100) GISLSSYLQSTQ(20/100) EHMALTYPFRPP(13/100) HWAPSMYDYVSW(7/100) RTVPDYTAHVRT(5/100) SGHQLLLNKMPN(4/100) TNSIWYTAPYMF(3/100) GMSLSRQMLWSL(2/100) HLFPQSNYGGHS(2/100) CIQLANPPRLXG(2/100) 10 Phage display (subtractive panning) 4 PMID:32417652 Invasive U87MG glioblastoma cell line Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phage library was first incubated with U87T cells to deplete the library of background phage. The supernatant was then incubated with U87R cells. 3541 VQHNTKYSVVIR(14)[7.04026] VQHNTKYSVVIR(10)[NA] HGTAPALKILK(2)[NA] HGTAPALKILK(1)[2.15974] LHHWNARSALAN(1)[NA] SSIEPAYYLHAT(1)[NA] EALNQWHIFLGR(1)[NA] HYKTPNLYKGWH(1)[NA] PSIFKHPKHLA(1)[NA] 9 Phage display (subtractive panning) 3 PMID:32040673 SUS316L stainless steel Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding ability of SBP-A and SBP-B on SUS316L surface was determined based on the amount of each biotinylated (Bio)-peptide binding on the SUS316L disk surface, as assessed by the relative SA-HRP activity measured by ABTS assay. Data shown were reproduced from Fig.2a in the reference. To remove phage particles non-specifically binding to the polystyrene dish, 5E+10 pfu of the phage library in phosphate buffered saline (PBS) (pH 7.4) containing 0.05% (w/v) Tween 20 (PBST) was added into a polystyrene dish. After 1 h incubation at room temperature with gentle agitation, the solution containing unbound phage was recovered and used as the primary library. After three rounds of phage panning procedure, 12 mer peptide (SBP-A; VQHNTKYSVVIR) was identified as SUS316L-binding peptide. The SBP-A peptide formed a stable bond to a SUS316L modified surface and was not toxic to HUVECs. The SBP-A was then used for anti-ICAM antibody modification on SUS316L to construct a vascular endothelial cell-selective surface. The constructed surface dominantly immobilized vascular endothelial cells to smooth muscle cells, demonstrating that the SBP-A enabled simple immobilization of biomolecules without disturbing their active biological function. 3542 VVSPDMNLLLTN(5/16)[0.912840±0.000310] VFSSMVHVLNTH(3/16)[0.140007±0.000179] SGVYKVAYDWQH(1/16)[0.235673±0.002483] GLHTSATNLYLH(1/16)[1.061523±0.002488] CYAGHDLYVAAD(1/16)[NA] SLSWLTKMQMEM(1/16)[NA] 6 Phage display (common panning) 3 PMID:31993846 Escherichia coli O157:H7 cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The color development was recorded using a microplate reader (Thermo Fisher, MultiskanTM GO) by monitoring absorbance at 405 nm. A405 shown was reproduced from Figure 2. Briefly, 10 μL phage library (~e13 pfu/mL) were added into 1 mL E. coli O157:H7 cell suspensions (OD600 = 0.5) and incubated for 1 h at room temperature with gentle agitation. Bacteria with bound phages were precipitated by spinning for 5 min at 16,000×g, and separated from unbound phages in solution by a series of 10 washing and centrifugation steps (16,000×g, 5 min) with 1 mL TBST buffer (50 mM Tris–HCl, 0.05% (v/v) Tween 20) each time. After washing, bound phages with E. coli O157:H7 were suspended in 200 μL elution buffer (0.2 M glycine–HCl, pH 2.2) with gentle shaking at room temperature for 10 min. The eluted phages were neutralized with 150 μL, 1 M Tris–HCl (pH 9.1). A 12-mer peptide (with the sequence of GLHTSATNLYLH) with high binding affinity toward Escherichia coli O157:H7 was discovered by biopanning of phage-displayed peptide library. The peptide modified with glycine residues (G3) and one cysteine (C) residue at C-terminal, could self-assemble on gold electrodes, enabling electrochemical impedance spectroscopy (EIS) analysis for quantitative detection of E. coli O157:H7. This method showed a low detection limit of 20 CFU/mL and a liner range from 2e2 to 2e6 CFU/mL. 3543 SGVYKVAYDWQH(6/16)[0.235673±0.002483] GLHTSATNLYLH(3/16)[1.061523±0.002488] VVSPDMNLLLTN(1/16)[0.140007±0.000179] GSAPLLTVDTSK(1/16)[NA] 4 Phage display (common panning) 4 PMID:31993846 Escherichia coli O157:H7 cells Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The color development was recorded using a microplate reader (Thermo Fisher, MultiskanTM GO) by monitoring absorbance at 405 nm. A405 shown was reproduced from Figure 2. Briefly, 10 μL phage library (~e13 pfu/mL) were added into 1 mL E. coli O157:H7 cell suspensions (OD600 = 0.5) and incubated for 1 h at room temperature with gentle agitation. Bacteria with bound phages were precipitated by spinning for 5 min at 16,000×g, and separated from unbound phages in solution by a series of 10 washing and centrifugation steps (16,000×g, 5 min) with 1 mL TBST buffer (50 mM Tris–HCl, 0.05% (v/v) Tween 20) each time. After washing, bound phages with E. coli O157:H7 were suspended in 200 μL elution buffer (0.2 M glycine–HCl, pH 2.2) with gentle shaking at room temperature for 10 min. The eluted phages were neutralized with 150 μL, 1 M Tris–HCl (pH 9.1). A 12-mer peptide (with the sequence of GLHTSATNLYLH) with high binding affinity toward Escherichia coli O157:H7 was discovered by biopanning of phage-displayed peptide library. The peptide modified with glycine residues (G3) and one cysteine (C) residue at C-terminal, could self-assemble on gold electrodes, enabling electrochemical impedance spectroscopy (EIS) analysis for quantitative detection of E. coli O157:H7. This method showed a low detection limit of 20 CFU/mL and a liner range from 2e2 to 2e6 CFU/mL. 3544 MHPNAGHGSLMR(8/19)[3.193117±0.007169] SGVYKVAYDWQH(8/19)[2.932297±0.006400] TGENHVADDRKN(1/19)[2.093027±0.008066] GLDGYRHPRDAW(1/19)[1.885507±0.010138] GLVDEGDHDGER(1/19)[2.386910±0.008071] 5 Phage display (subtractive panning) 3-4 PMID:31270859 Matrix metalloproteinase-9, MMP-9 Integrin beta-5 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Binding of the phages on MMP-9 and BSA was examined with phage ELISA. The absorbance was measured on a Multiskan ELISA reader at 450nm. The binding ratio was calculated and reproduced from Figure 3 in the reference. For this process, we added 1e11 pfu phages from the M13 phage display peptide library (Ph.D.‐12) (New England Biolabs, Herts, UK) to 2 μg of active human recombinant MMP‐9 in the first round and incubated this mixture for 2 hours in 200 μL of tris‐buffered saline with Tween 20 (TBST) buffer (0.1% Tween 20). After that, we added 50 μL of Ni magnetic beads (His60 Ni Magnetic Beads, Takara Bio Inc, Kusatsu, Shiga Prefecture, Japan) to the mixture and incubated that for another 2 hours. The supernatant containing unbound phage was removed via magnetic separation. By elution magnetic beads 10 times with 1 mL of TBST buffer (0.1% Tween 20), we removed feeble or nonspecifically bound phage. In the end, according to the guideline of the manufacturer the bound phage particles were recovered via infecting Escherichia coli ER2738 host. We also preincubated one portion of the first round amplified phage with the beads in the lack of target to eliminate nickel magnetic beads-specific phage particles. By applying magnetic fishing, the unbound phage fraction was incubated with recombinant human MMP‐9 and specifically bound phages were separated. Interestingly, in silico molecular docking showed strong interactions between the peptide threedimensional models and some important residues of the MMP-9 and MMP-2 proteins at the fibronectin domain. A consensus peptide sequence (MHPNAGHGSLMR) was then synthesized (named as RSH-12) to evaluate its inhibitory potency by in vitro assays. Zymography assay was employed to evaluate the effect of RSH-12 on gelatinolysis activity of MMP-2 and MMP-9 secretion from the HT1080 cells using different concentrations of RSH-12 and inhibiting MMP-9- and MMP-2- driven gelatin proteolysis, measured by fluorescein isothiocyanate-gelatin degradation assay and HT1080 cell invasion assay on Matrigel (gelatinous protein mixture). The negative control peptide (CP) with the irrelevant sequence and no MMP inhibition properties and the positive control compound (GM6001) as a potent inhibitor of MMPs were used to assess the selectivity and specificity of gelatinases inhibition by RSH-12. Therefore, RSH-12 decreased the gelatin degradation by specifically preventing gelatin binding to MMP-9 and MMP-2. 3545 WHWRLWDVPDNP(22)[2.081931±0.454971] SFVNLWTPRYSL(5)[0.777530±0.359939] TLFSKPYPNSSR(1)[0.907007±0.005364] TPMHYPATPSPH(1)[1.105431±0.004797] QFGPVFTWLNHA(1)[4.510443±0.010535] TITNAPIKDLTP(1)[0.680020±0.007786] LTPHKHHKHLHA(1)[0.93332] DPHGSLFPRTHP(1)[1.945993±0.003895] TQYPIDGDIFRR(1)[0.999870±0.007093] HLTWIPSVVRNS(1)[0.278147±0.003574] WHWAWYSPTARM(1)[2.754893±0.008042] 11 Phage display (subtractive panning) 3 PMID:32737386 Non-structural protein 1, NS1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Optical density (OD) was measured at 450 nm (measurement wavelength) and 620 nm (reference wavelength) using a Synergy H1 multi-mode microplate reader (Biotek Instruments, Winooski, VT, USA). The OD450 was reproduced from Figures 2a and 2b in the reference and shown. The phage-displayed peptide library ( e11 pfu) was incubated in the BSA-coated wells at RT for 1 h to deplete BSAbinding phages. Unbound phages from the BSA-coated wells were transferred to the DENV NS1-coated wells and incubated at RT for 1 h. The DENV NS1-coated wells were then thoroughly washed to remove unbound phages. DENV NS1-binding phages were eluted from the wells by incubating with glycine elution buffer (0.2 M glycine–HCl pH 2.2 and 1 mg/ml BSA) at RT for 20 min, followed by neutralization with 1 M Tris–HCl pH 9.1. The eluted phages were amplified in E. coli ER2738 cultures and titrated according to the manufacturer’s protocol with minor modifications. The amplified phages ( e11 pfu) from the first round were used for the next round of biopanning, and a total of 3 rounds of biopanning were performed. The number of phages obtained from each round was titrated to evaluate the efficiency of phage recovery. We performed biopanning assays using a phage-displayed peptide library and identified 11 different sequences of 12-mer peptides binding to DENV NS1. In silico analyses of peptide-protein interactions revealed 4 peptides most likely to bind to DENV NS1 at specific positions and their association was analysed by surface plasmon resonance. Treatment of Huh7 cells with these 4 peptides conjugated with N-terminal fluorescent tag and C-terminal cell penetrating tag at varying time-of-addition post-DENV infection could inhibit the production of DENV-2 in a time- and dose-dependent manner. The inhibitory effects of the peptides were also observed in other virus serotypes (DENV-1 and DENV-4), but not in DENV-3. These findings indicate the potential application of peptides targeting DENV NS1 as antiviral agents against DENV infection. 3546 NVFTVSP(6)[0.816657±0.001570, 12.09] NSLSISY(3)[0.371417±0.001570, NA] SQQGKLN(2)[0.525270±0.002944, NA] ELTPLTL(2)[0.182410±0.003243, NA] QLAVAPS(2)[0.255427±0.001570, NA] DRLSHTR(2)[0.252820±0.002944, NA] QTLNVKP(2)[0.215853±0.001570, NA] VVTPKTA(2)[0.220397±0.001570, NA] TTQVLEA(2)[0.208373±0.000981, NA] NLLDSLH(1)[0.208033±0.001570, NA] WSLSELH(1)[0.319490±0.002720, NA] 11 Phage display (common panning) 3 PMID:31805343 Fibroblast growth factor 9, FGF-9 FGFR-1, FGFR-2, FGFR-3 and FGFR-4 Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The binding activity of the 11 positive phage clones to FGF9 was verified by ELISA assay. The absorbance was measured at 450 nm. The OD450 value (in the first column) was reproduced from Figure 1A and shown. In addition, to confirm the interaction between P4 peptide and FGF9, the surface plasmon resonance (SPR) on the immobilized FGF9 was applied to determine the affinity constants. The binding kinetics of the peptides to FGF9 was detected by OpenSPR™ (Nicoya Lifesciences, Waterloo, Canada). The Kd (μM) value was shown in the second column of the affinity values. A sterile polystyrene petri dish was coated with 10 μg/ml recombinant human FGF9 or PBS at 4 °C in 0.1 M NaHCO3 overnight, and blocked with bovine serum albumin (BSA) at 5 mg/ml in 0.1 M NaHCO3 for 2 h at room temperature. After the dish was washed five times (1 min each) with 0.05 % PBST (PBS containing 0.05 % Tween-20), the diluted original Ph.D.-7 library (2.0e11) was added and incubated for 2 h at room temperature. The unbound phages were removed through washing ten times (1 min each) with 0.05 % PBST prior to elution of the bound phages with continuous shaking in 1 ml of 0.1 M glycine-HCl for 10 min at room temperature and subsequent neutralization with 100 μl of 1 M Tris-HCl. The eluate was amplified and purified for the next round of screening. Two additional rounds of selection were conducted under more rigorous conditions. In short, dishes were coated with less amount of FGF9 (5 μg/ml and 2.5 μg/ml for the second and third round, respectively), incubated with the previous eluate for shorter time (1.5 h and 1 h for the second and third rounds, respectively), and washed with higher concentration of PBST for a longer time (0.1 % PBST for 10 × 2 min and 0.3 % PBST for 10 × 3 min for the second and third round, respectively). After three successive rounds of strict selection, the phage clones obtained by infection of E. coli ER2738, individual selection and amplification were exposed to Enzyme-linked immunosorbent assay (ELISA). Sequence comparison showed that P4 (NVFTVSP) shared high homology with the conserved motif in the immunoglobulin-like (Ig-like) domain II∼III (D2-D3) linker of the FGF9 high-affinity receptor (FGFR3c). The interaction between P4 and FGF9 was confirmed by the surface plasmon resonance (SPR) assay. Functional analysis indicated that P4 counteracted FGF9-induced aggressive phenotype, including cell proliferation, migration, and invasion in vitro, as well as suppressed tumor growth in vivovia down-regulation of the MAPKs and Akt cascades. More importantly, we found that FGF9 served as an underlying mechanism of the chemoresistance in GC and BC cells, and P4 could increase the sensitivity to the chemical agent via antagonizing the suppression effects of FGF9 on cell apoptosis. Taken together, our study identified a novel binding peptide for FGF9, which may serve as a potential therapeutic agent for malignant tumors featured by abnormally up-regulation of FGF9. 3547 LPHWHPHSHLQP(42.9%) RQRPKDHFFSRP(14.3%) SPVMPFSPYSTW(14.3%) VPWYKHPRHPHL(14.3%) FHKHKNPGSPII(7.1%) FHRHHSPPPSII(7.1%) 6 Phage display (common panning) 5 PMID:31289973 Silica, Si Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The phages binding on the silica wafer were eluted by incubating with 200 μL of glycine–HCl solution (0.2 M, pH 2.2) at room temperature for 10 min. After neutralization by adding 30 μL of 1 M Tris–HCl (pH 9.1), eluted phages were amplified in E. coli ER2738, and better affinity-binding phages were obtained after the first round of biopanning. After that, the amplified phages were incubated with the silica wafer to perform the above biopanning round by round. A total of five rounds of biopanning were conducted and the silica affinity-binding phages were enriched. The optimal 12-mer SAP sequence that specifically and intensively binding to silica through affinity interaction was obtained by DNA sequencing of the SAP selected from round-5 biopanning phages. The optimal screened SAP (LPHWHPHSHLQP) was selected from a M13 phage display peptide library and fused to the C-terminal of DAAO to obtain fused DAAOs with one, two and three SAP tags, respectively. The activity of DAAO-SAP-MPS was superior comparing with DAAO-2SAP-MPS and DAAO-3SAP-MPS; meanwhile DAAO-SAP-MPS shows 36% higher activity than that of DAAO-MPS. Fusion with one SAP improved the thermal stability with a 10% activity increase for immobilized DAAO-SAP-MPS compared to that of DAAO-MPS at 50 °C for 3 h. Moreover, the activity recovery of immobilized DAAO-SAP-MPS was 25% higher in operation stability assessment after six-batch conversions of cephalosporin to glutaryl-7-amino cephalosporanic acid than that of DAAO-MPS. 3548 EIAYPARYANTY IPWTQHMAMSPM QNKLWDTPSNPW TALGHQPLMRNT SGGMPTARMSHQ APWHNSWSEERT ESGLWYSIDMKP YLDEFAWYRFTH WPRPYYGDWFQT YPPPDSHSERVE SMQGKAYGGTVM WPRPYYGEGFQT HPLTWNLRSSPA ADWYHWRSHSSS VVSPDMNLLLTN TSLDGRISYHNR VHWDFRQWWQPS SQWETSQMIQKM LPLTRGYVGDQY FSPHADWVVVSG GFAVGARDSLMF 21 Phage display (subtractive panning) 3 PMID:30789148 Sera from 30 Systemic lupus erythematosus (SLE) patients Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Serum pools were obtained from 30 HCs and 30 SLE patients. Microtitre wells were coated overnight at 4°C with 150 μl sera from healthy controls diluted with NaHCO3. Plates were blocked with 3% non-fat milk for 2 h at 37°C, and subsequently washed six times with phosphate-buffered saline with Tween 20 (0.05% Tween 20). A 100-μl diluted random 12-peptide phage display library with a titre of 2.0e11 pfu/ml was added to the coated plates. After incubation for 1 h at room temperature, unbound phages were collected and 100μl was added per well to SLE serumcoated plates. Following incubation for 1 h at room temperature, bound phages were eluted with 100 μl 0.2 mol/l glycine–HCl (pH 2.2) and neutralised with 1 mol/l Tris-HCl (pH 9.1). Eluted phages were amplified in a host strain and purified by precipitation for about 4 h using one-sixth volume of polyethylene glycol/NaCl. Another two rounds of SLE-serum affinity selection were carried out using the same process. After a negative selection step with serum from healthy controls (HCs), a phage library of 12 peptides was used for three rounds of screening with sera from 30 SLE patients. After four rounds of biopanning, 21 positive peptides were sequenced. We produced 37-feature arrays containing 16 recombinant citrullinated peptides. The microarrays were tested with an independent validation set of serum samples from 50 HCs, 60 SLE patients, and 60 rheumatoid arthritis (RA) patients. 3549 SQSGPTIFYNLA(13)[2.203160±0.009388] AESRRPFFEPFM(2)[2.833720±0.009388] FNPSFAKMGNSL(1) SALTSAVSGSAA(1) 4 Phage display (common panning) 3 PMID:32808238 Anti-epstein-barr virus (EBV) multiple sclerosis(MS) brain IgG antibodies (MS 95–2) Epstein-barr virus (EBV) Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Purified MS brain 95–2 IgG in 0.1 M carbonate buffer (50 μl, 200 μg/ml) was coated onto wells of ELISA plates overnight at 4 °C. The wells were then blocked with 3% BSA for 2 h followed by incubation with purified phage 3–2 and 3–3 (5 × 109/well) for 1 h. After washing with 0.05% Tween-20/Tris buffered saline (TBST), the wells were incubated with a 1:500 dilution of mouse anti-M13 IgG-HRP (New England BioLabs) antibody for 1 h, followed by incubation with peroxidase substrate ABTS (Vector Laboratories, Burlingame, CA) for 20 min. The optical absorbance was measured at 415 nm with a Microplate Manager (BioRad, Hercules, CA). All samples were tested in duplicate and the ELISA was repeated at least one additional time. OD415 were reproduced from Figure 3A and shown. Wells were blocked with 3% BSA in Tris-buffed saline (TBS) for 2 h, washed 6 times with 0.1% Tween20–TBS (TBST), and the phage peptide library (1.5e11 phage in 100 μl of TBST) was added to the wells and incubated for 2 h at room temperature. Wells were washed 20–30 times with TBST, and bound phage were eluted with 100 μl of 0.2 M glycine, pH 2.2/0.1% BSA for 10 min. The phage-containing solution was immediately neutralized with 15 μl of 1 M Tris, pH 9.5, and eluted phage were titered and reamplified. Amplified phage (e11 pfu) from the first round of panning was applied to additional rounds of panning using the same conditions as in the first pan, except that the concentration of Tween20 in the washing buffer was increased to 0.5%. After the third round of panning, affinity-selected phage were titered. Single plaques were amplified and the peptides were sequenced. We screened phage-displayed random peptide libraries (12-mer) with total IgG antibodies purified from the brain of a patient with acute MS. We identified and characterized the phage peptides for binding specificity to intrathecal IgG from patients with MS and from controls by ELISA, phage-mediated Immuno-PCR, and isoelectric focusing. We identified two phage peptides that share sequence homologies with EBV nuclear antigens 1 and 2 (EBNA1 and EBNA2), respectively. The specificity of the EBV epitopes found by panning with MS brain IgG was confirmed by ELISA and competitive inhibition assays. 3550 TPDCTRWWCPLT YPICTDTLCRLS RQRTPPTRTIRS RPRTRLHTHRNR TQTTKSISTNRI 5 Phage display (common panning) 5 PMID:31812636 Recombinant envelope (E) protein E-TrX Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA After five rounds of bio-panning, phages showing high binding activity to the recombinant E protein based on an indirect enzyme-linked immunosorbent assay (ELISA) were selected (data not shown). The Ph.D.-12™ Phage Display Peptide Library (New England Bio labs, USA) was exposed to plates coated with purified recombinant protein E-Trx. Unbound phages were washed away, and specifically bound phages were eluted and amplified for a second set of screening. After five rounds of bio-panning, phages showing high binding activity to the recombinant E protein based on an indirect enzyme-linked immunosorbent assay (ELISA) were selected then sequenced. P1(TPDCTRWWCPLT) inhibits JEV infection in BHK-21 cells with 50% inhibitory capacity at a concentration of 35.9 μM. The time-of-addition assay indicates that JEV replication is significantly inhibited during pre-infection and co-infection of P1 with JEV while post-infection treatments with P1 have very little impact on JEV proliferation, showing that P1 inhibits JEV infection at early stages and indicating the potential prophylactic effect of P1. 3551 SVSVGMKPSPRP 1 Phage display (common panning) 5 PMID:31812636 Recombinant envelope (E) protein TrX Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA After five rounds of bio-panning, phages showing high binding activity to the recombinant E protein based on an indirect enzyme-linked immunosorbent assay (ELISA) were selected (data not shown). The Ph.D.-12™ Phage Display Peptide Library (New England Bio labs, USA) was exposed to plates coated with purified recombinant protein Trx. Unbound phages were washed away, and specifically bound phages were eluted and amplified for a second set of screening. After five rounds of bio-panning, phages showing high binding activity to the recombinant E protein based on an indirect enzyme-linked immunosorbent assay (ELISA) were selected then sequenced. 3552 AGDVPRGWTSSS HTPAATLHPVFL TPSQSMGWDSSA THNKVQQ NTTLNGL PETWTHW TNHOKTW DYRVQMA HQSQSRM PTKWSAT PNSVKAQ GLELREK 12 Phage display (in vivo) 3 PMID:31589023 C57bl/6 mice heart Not determined. Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool For library screening, Ph.D.−12 and C7C phage library (New England Biolabs, USA) were mixed, 1.5e13 pfu/mL, and injected intraperitoneally (IP) into C57bl/6 mice (100 μl, n=2 per group). Prior to tissue harvest, mice were euthanized and perfused with 20 mL saline to ensure complete tissue exsanguination. Spinal cord, brain, and cardiac tissue was harvested and flash-frozen in DMEM (Invitrogen, Inc.) with 1% BSA (Fisher Scientific). After homogenization, tissue homogenates were centrifuged and resuspended in 1% Triton-X 100 (Sigma-Aldrich, Inc.) and passed through a 25-gauge needle to ensure cell lysis. Phage titer was quantified and spinal- and brain-tissue homogenate was amplified for subsequent screens. The series of TACL peptides were synthesized and tested for their ability to deliver a model enzyme (NeutrAvidin-horseradish peroxidase fusion) to the brain and spinal cord. Three TACL-peptides facilitated significant active enzyme delivery into the CNS, with limited accumulation in off-target organs. Peptide structure and serum stability is increased when internal cysteine residues are cyclized by perfluoroarylation with decafluorobiphenyl, which increased delivery to the CNS further. TACL-peptide was demonstrated to localize in parasympathetic ganglia neurons in addition to neuronal structures in the hindbrain and spinal cord. By targeting uptake into ANS neurons, we demonstrate the potential for TACL-peptides to bypass the blood-brain barrier and deliver therapeutics into the brain and spinal cord. 3553 NIVCPNEHPRCS THNKVQQ DYRVQMA PSHLTKM SQKNFTH PLSKGKL 6 Phage display (in vivo) 3 PMID:31589023 C57bl/6 mice brain Not determined. Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool For library screening, Ph.D.−12 and C7C phage library (New England Biolabs, USA) were mixed, 1.5e13 pfu/mL, and injected intraperitoneally (IP) into C57bl/6 mice (100 μl, n=2 per group). Prior to tissue harvest, mice were euthanized and perfused with 20 mL saline to ensure complete tissue exsanguination. Spinal cord, brain, and cardiac tissue was harvested and flash-frozen in DMEM (Invitrogen, Inc.) with 1% BSA (Fisher Scientific). After homogenization, tissue homogenates were centrifuged and resuspended in 1% Triton-X 100 (Sigma-Aldrich, Inc.) and passed through a 25-gauge needle to ensure cell lysis. Phage titer was quantified and spinal- and brain-tissue homogenate was amplified for subsequent screens. The series of TACL peptides were synthesized and tested for their ability to deliver a model enzyme (NeutrAvidin-horseradish peroxidase fusion) to the brain and spinal cord. Three TACL-peptides facilitated significant active enzyme delivery into the CNS, with limited accumulation in off-target organs. Peptide structure and serum stability is increased when internal cysteine residues are cyclized by perfluoroarylation with decafluorobiphenyl, which increased delivery to the CNS further. TACL-peptide was demonstrated to localize in parasympathetic ganglia neurons in addition to neuronal structures in the hindbrain and spinal cord. By targeting uptake into ANS neurons, we demonstrate the potential for TACL-peptides to bypass the blood-brain barrier and deliver therapeutics into the brain and spinal cord. 3554 GFPSVRDLSPLR WATLDLGPQPYS PETWTHW HQSQSRM PLSKGKL NTSPLNT PTKWSAT ASGFTAT PNSVKAQ TMSTTQV GLELREK 11 Phage display (in vivo) 3 PMID:31589023 C57bl/6 mice spinal cord Not determined. Not determined. Not determined. Ph.D.-12 and Ph.D.-C7C phage display library pool For library screening, Ph.D.−12 and C7C phage library (New England Biolabs, USA) were mixed, 1.5e13 pfu/mL, and injected intraperitoneally (IP) into C57bl/6 mice (100 μl, n=2 per group). Prior to tissue harvest, mice were euthanized and perfused with 20 mL saline to ensure complete tissue exsanguination. Spinal cord, brain, and cardiac tissue was harvested and flash-frozen in DMEM (Invitrogen, Inc.) with 1% BSA (Fisher Scientific). After homogenization, tissue homogenates were centrifuged and resuspended in 1% Triton-X 100 (Sigma-Aldrich, Inc.) and passed through a 25-gauge needle to ensure cell lysis. Phage titer was quantified and spinal- and brain-tissue homogenate was amplified for subsequent screens. The series of TACL peptides were synthesized and tested for their ability to deliver a model enzyme (NeutrAvidin-horseradish peroxidase fusion) to the brain and spinal cord. Three TACL-peptides facilitated significant active enzyme delivery into the CNS, with limited accumulation in off-target organs. Peptide structure and serum stability is increased when internal cysteine residues are cyclized by perfluoroarylation with decafluorobiphenyl, which increased delivery to the CNS further. TACL-peptide was demonstrated to localize in parasympathetic ganglia neurons in addition to neuronal structures in the hindbrain and spinal cord. By targeting uptake into ANS neurons, we demonstrate the potential for TACL-peptides to bypass the blood-brain barrier and deliver therapeutics into the brain and spinal cord. 3555 CWRDYLI(10)[8.2±0.8] CQWFSHR(8)[6.9±0.9] CGTWLKF(7)[NA] 3 Phage display (subtractive panning) 3 PMID:31398275 Tobacco Etch Virus protease (TEV protease) Not determined. Not determined. Not determined. pADLg3-TGC-(NNK)6-TAG phagemid library Fluorescence polarization We incubated a 25 nM 5-FAM-conjugated cyclic peptide and different concentrations of a target protein (160 nM to 160 µM) at black 96-well plates in a 200 µL total volume that was adjusted by adding the PBS buffer and then measured fluorescence polarization in a microplate reader at Ex/Em = 490 nm/520 nm. The Kd (μM) value was determined and shown. In the selection, in order to remove individuals capable of non-specific binding we incubated the phage library with only streptavidin magnetic beads for every round of selection, collected the unbound phages, and then subjected them to bind protein-binding streptavidin magnetic beads. After the third round, we sequenced 25 phage clones that converged to only three peptide sequences, CWRDYLI-AcrK, CQWFSHR-AcrK, and CGTWLKF-AcrK. The results indicated that both CWRDYLI-Ark and CQWFSHR-Ark bind to TEV protease with a single digit μM dissociation constant and both cyclic peptides bind to TEV protease significantly better than their linear counterpart (>6-fold). 3556 CQSLWMN(8)[7.1±0.7,9.7±0.7] CKHSLWV(2)[NA, NA] CLSDCRV(1)[NA, NA] 3 Phage display (subtractive panning) 3 PMID:31398275 Histone deacetylase 8, HD8 Not determined. Not determined. Not determined. pADLg3-TGC-(NNK)6-TAG phagemid library Fluorescence polarization We incubated a 25 nM 5-FAM-conjugated cyclic peptide and different concentrations of a target protein (160 nM to 160 µM) at black 96-well plates in a 200 µL total volume that was adjusted by adding the PBS buffer and then measured fluorescence polarization in a microplate reader at Ex/Em = 490 nm/520 nm. The Kd (μM) value was determined and shown in the firt column of the affinity values. In addition, IC50 (μM) values of selected cyclic peptides when binding to their protein targets were also determined and shown in the second column of the affinity values. In the selection, in order to remove individuals capable of non-specific binding we incubated the phage library with only streptavidin magnetic beads for every round of selection, collected the unbound phages, and then subjected them to bind protein-binding streptavidin magnetic beads. All selected cyclic peptide ligands showed 4 to 6-fold stronger affinity to their protein targets than their linear counterparts. 3557 LLADTTHHRPWT[0.472097±0.001114,0.32025] HATGTHGLSLSH[0.074187±0.001120,0.118277±0.001120] TSGYLHLRGRWR[0.03526,0.0615] 3 Phage display (subtractive panning) 4 PMID:31786302 C-terminal domain of CCN family member 2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Positive phage clones were adsorbed to PKW-CTGF- and TrxA-CTGF-coated microplates, and the optical density at 450 nm (OD450) was directly determined using ELISA. OD450 values were reproduced from Figure 2A and shown. The first column of affinity values was the binding affinity of phage clones to PKW-CTGF, and the second column the binding affinity of phage clones to TrxA-CTGF. Human TrxA-CTGF/C and TrxA dissolved in sodium bicarbonate (NaHCO3, pH 8.6) were coated onto 96-well plates and incubated overnight at 4 °C. After blocking with 0.5% (w/v) bovine serum albumin (BSA), the wells were washed with TBST 6 times. The random peptide library was added to the TrxA-coated plate and incubated at 37 °C for 1 h. Supernatants were then added to the TrxA-CTGF/C-coated plate for 1 h at 37 °C. The plates were then washed six times with TBST and eluted with 0.2 M glycine/HCl (pH 2.2) containing 0.5% BSA, then immediately neutralized with Tris/HCl (pH 9.1). Next, phages were inoculated into LB medium with 10% E2738 at 37 °C for 4.5 h and centrifuged at 30,000 ×g for 15 min at 4 °C, the supernatants added to 1/6 volume PEG8000/NaCl at 4 °C overnight. Four rounds of selection were performed as follows: the supernatants were centrifuged at 12,000 ×g for 15 min and suspended in 0.5% Tween20-TBST; in the second round washed 10 times 0.5% Tween20-TBST; in the third round, washed 15 times with 1% Tween20-TBST; and, finally, in the fourth round, washed 20 times with 2% Tween20-TBST. 810A (LLADTTHHRPWT) was developed as a novel peptide, which binds to the C-terminal domain of connective tissue growth factor (CTGF). It could effectively inhibit the proliferation, migration, and expression of TGF-β and α-SMA in cells pretreated with CTGF. In in vivo experiments using an animal model of BLM-induced pulmonary fibrosis, the peptide demonstrated similar effects, which effectively reduced the onset and development of pulmonary fibrosis in mice. 3558 WEYDRYRGWHIG(2) RWPPHFEWHFDD(1) 2 Phage display (common panning) 4 PMID:32176482 Anti-F1 capsular antigen monoclonal antibody (YPF19) and human sera pool IgG of Alzheimer’s disease (AD) patients F1 capsule antigen Not determined. Not determined. pVIII-12aa phage display library (X12) ELISA Immunoscreening:positive spots on the immunoblots were detected using the Stable DAB chromogen system (Life Technologies, Monza, Italy). ELISA:the ELISA signal was measured at 450 nm using Labsystem Multiskan Bichromatic. Briefly, in the first round of selection, 500 μL of DYN−mAb YPF19 were incubated with 100 μL of each of the four phage libraries with a titer of e12 for 3−4 h at room temperature under mild stirring. The beads were washed 3 times in PBS−0.05% Tween 20 and separated with a magnetic device for 1−2 min to eliminate supernatants containing phage that did not bind the target present on the functionalized beads. Selected phage clones were eluted from antibodies with 500 μL of eluting buffer, 0.2 M of glycine-HCl (pH 2.2) + 0.1% BSA, neutralized immediately with 1 M Tris−HCl, pH 9.6. The enriched phage pools were amplified by infecting TG1 E. coli, purified twice by PEG precipitation, titrated, and used as the input for further panning. Biopanning affinity selection was repeated in the second round against DYN−IgG-AD, then in the third round against DYN−mAb YPF19 again, and finally a fourth selection round was carried out as the second one. Peptide 12III1(RWPPHFEWHFDD), was found to be able to prevent in vitro Aβ1-42-induced cytotoxicity in SH-SY5Y cells, as well as to promote disaggregation of preformed fibrils, to a greater extent with respect to wild-type phage (pC89). IgG levels detected by 12III1 provided a significant level of discrimination between diseased and nondemented subjects, as well as a good correlation with the state progression of the disease. These results give significant impact in AD state and stage diagnosis, paving the way for the development not only for an innovative blood diagnostic assay for AD precise diagnosis, progressive clinical assessment, and screening but also for new effective treatments. 3559 GGGCIEGPCLEG(2) WVGCHGEWCGVW(1) HRGCIEGPCLDA(1) 3 Phage display (common panning) 4 PMID:32176482 Anti-F1 capsular antigen monoclonal antibody (YPF19) and human sera pool IgG of Alzheimer’s disease (AD) patients F1 capsule antigen Not determined. Not determined. pVIII-12aa-Cys phage display library (X3CX4CX3) ELISA Immunoscreening:positive spots on the immunoblots were detected using the Stable DAB chromogen system (Life Technologies, Monza, Italy). ELISA:the ELISA signal was measured at 450 nm using Labsystem Multiskan Bichromatic. Briefly, in the first round of selection, 500 μL of DYN−mAb YPF19 were incubated with 100 μL of each of the four phage libraries with a titer of e12 for 3−4 h at room temperature under mild stirring. The beads were washed 3 times in PBS−0.05% Tween 20 and separated with a magnetic device for 1−2 min to eliminate supernatants containing phage that did not bind the target present on the functionalized beads. Selected phage clones were eluted from antibodies with 500 μL of eluting buffer, 0.2 M of glycine-HCl (pH 2.2) + 0.1% BSA, neutralized immediately with 1 M Tris−HCl, pH 9.6. The enriched phage pools were amplified by infecting TG1 E. coli, purified twice by PEG precipitation, titrated, and used as the input for further panning. Biopanning affinity selection was repeated in the second round against DYN−IgG-AD, then in the third round against DYN−mAb YPF19 again, and finally a fourth selection round was carried out as the second one. 3560 CQGPTFKCDAIWREC(20)[3.4±0.2] CVPQPDTCAKIDPRC(8)[>300] CLLQQPVCGEWNPKC(7)[>300] CYAAQGGCLRDWTRC(4)[~100] CYAAQGGCLRDWTRC(3)[>300] CYAINQRCLQEWSRC(3)[~250] CFADGSNCVVEWSEC(3)[N.D.] CIVQQGLCHEWNPRC(3)[>300] CVWLQATCTRSWSGC(3)[~300] CRPQPDTCVSLSGEC(2)[>300] CVPQPDTCANMDPEC(1)[>300] CKPQPDTCLHTTGKC(1)[N.D.] CYAAKGGCQPNWTQC(1)[~100] CYQSWPVCQAWNPRC(1)[>300] CQHAGAVCHWWNPRC(1)[>300] CNQDLMVCRFWNSRC(1)[>300] CRKMAEDCGWGVLVC(1)[>300] CKKVSMSCGWGEAVC(1)[>300] CRERMAVCGWGVQVC(1)[>300] CNWQTQVCVRDWLGC(1)[~300] CSWQQGECTRTWGGC(1)[>300] 21 Phage display (common panning) 3 PMID:31251434 72 kDa type IV collagenase Not determined. Not determined. Not determined. CX6CX6C phage display library Fluorescence The inhibitory activity of bicyclic peptides was determined by measuring sidual protease activity with a fluorogenic substrate. For the FS-6 substrate, excitation was measured at 325 nm and emission at 400 nm. For the OMNIMMP® RED substrate, excitation was measured at 545 nm and emission at 576 nm. The inhibition constants (Ki) were calculated according to the equation of Cheng and Prusoff Ki = IC50/(1+([S]0/Km) wherein IC50 is the peptide concentration at which 50% of the enzymaticactivity is inhibited, [S]0 is the total substrate concentration, and KM is the Michaelis-Menten constant. Ki (μM) values were shown. Bacterial cells of a phage library glycerol stock were added to a 2 L falcon flask containing 500 ml of 2YT/chloramphenicol (30 μg/ml) medium to obtain an OD600 of 0.1. The culture was shaken (200 rpm) for 16 hr at 30°C. Bacterial cells were pelleted by centrifugation at 16,000 g and 4°C for 30 min, and the phage in the supernatant were purified by PEG-precipitation as follows. Briefly, 0.2 volumes of PEG solution (20% (w/v) polyethylene glycol 6000, 2.5 M NaCl) was added to the supernatant, mixed, incubated on ice for 30 min, and centrifuged at 2,700 g and 4°C for 30 min. PEG-purified phage, typically 1011–1012 t.u. (transducing units) were resuspended in 20 ml of 20 mM NH4HCO3, 5 mM EDTA, pH 8.0. Disulfide bridges were reduced by addition of 1 mM of TCEP and incubation at 42°C for 1 hr. The concentration of TCEP was subsequently reduced by repetitive concentration and dilution steps with reaction buffer (20 mM NH4HCO3, 5 mM EDTA, pH 8.0, degassed) in a Vivaspin-20 filter (MWCO of 10,000, Sartorius-Stedim Biotech GmbH). The volume of the phage solution was adjusted to 32 ml with reaction buffer and 8 ml of 50 μM tris-(bromomethyl)benzene (TBMB) in acetonitrile (MeCN) were added to obtain a final TBMB concentration of 10 μM. The reaction was incubated at 30°C for 1 hr before non-reacted TBMB was removed by precipitation of the phage with 0.2 volumes of 20% (w/v) polyethylene glycol 6000, 2.5 M NaCl on ice and centrifugation at 2,700 g and 4°C for 30 minutes. The phage pellet was resuspended in 3 ml of washing buffer (10 mM Tris-Cl, pH 7.4, 150 mM NaCl, 10 mM MgCl2, 1 mM CaCl2). Biotinylated active MMP-2 (180 μg) was added to 50 μl magnetic streptavidin beads (Dynabeads M-280 from Invitrogen Dynal Biotech AS) in washing buffer and incubated on a rotating wheel for 10 min at room temperature (RT). The magnetic beads were then washed with 0.5 ml washing buffer and incubated for 30 min at RT with 0.5 ml washing buffer containing 1% (w/v) BSA and 0.1% (v/v) Tween 20. At the same time the chemically modified phage (typically e10–e11 t.u. dissolved in 3 ml of washing buffer) were blocked by addition of 1.5 ml of washing buffer containing 3% (w/v) BSA and 0.3% (v/v) Tween 20 for 30 minutes. The blocked beads/target protein mixture (0.5 ml) and phage (4.5 ml) were mixed together and incubated for 30 minutes on a rotating wheel at room temperature. The beads were washed eight times with washing buffer containing 0.1% (v/v) Tween 20 and twice with washing buffer. The phage were eluted by incubation with 100 μl of 50 mM glycine, pH 2.2 for 5 minutes (pH-dependent elution), and then transferred to 50 μl of 1 M Tris-Cl, pH 8.0 for neutralization. The eluted phage were incubated with 30 ml of TG1 cells at OD600 of 0.4 for 90 minutes at 37°C, and the cells were plated on large 2YT/chloramphenicol (30 μg/ml chloramphenicol) plates. The second and third round of panning were performed following the same procedure but using in the second round neutravidin-coated magnetic beads instead of streptavidin to prevent the enrichment of streptavidin-specific peptide binders. Neutravidin beads were prepared by reacting 0.8 mg neutravidin (Pierce) with 0.5 ml of tosyl-activated magnetic beads (2e9 beads/ml; Dynabeads M-280, Invitrogen Dynal Biotech AS) according to the supplier's instructions. The abundant peptide M21 (ACQGPTFKCDAIWRECG) inhibited MMP-2 with a Ki of 3.4±0.2 μM. An alanine scan of M21 revealed that the first four amino acids of the C-terminal ring, Asp-Ala-Ile-Trp, were most important for the binding. 3561 WIPNSEFEHERT(23/50)[2.0] 1 Phage display (subtractive panning) 3 PMID:31301216 Fab A Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Fluorescence polarization The labeled peptide was diluted in PBS to prepare a stock solution of 500 nM. Fab A was diluted in PBS to two different concentrations (2 and 20 μM). Concentrations of peptide and Fab A were measured using a NanoDrop UV Spectrophotometer (Harlow Scientific). The level of binding was evaluated using fluorescence polarization (FP) by mixing equal volumes (20 μL) of the peptide and each Fab concentration in a 384 well black plate, resulting in a final peptide concentration of 250 nM. The dissociation constants (KD, μM) were determined. Three positive selections with Fab A as target and two negative selections were performed. For the first two positive rounds, Fab A was bound to protein L functionalized beads, while the last positive round employed covalent immobilization of the Fab on NHS beads. The negative selections were included to eliminate nonspecific binding of the phages to plastic, protein L, magnetic beads, and/or BSA. The first negative selection was performed by incubating with a BSA blocked tube containing protein L magnetic beads after the first positive panning rather than before, to avoid losing promising phage candidates that may have possessed significant binding to the target. The second negative selection was performed after the second positive selection, equal to the first one. Peptide B1 (WIPNSEFEHERT) was shown to selectively recognize Fab A. B1 having the lowest dissociation constant (KD) of 2 μM to a single Fab target. 3562 WHYNWQDVSDRQ[4.1] HQNHHSTFWEIY[6.5] 2 Phage display (subtractive panning) 4 PMID:31301216 Fab A,Fab Z and Fab D Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Fluorescence polarization The labeled peptide was diluted in PBS to prepare a stock solution of 500 nM. Fab A was diluted in PBS to two different concentrations (2 and 20 μM). Concentrations of peptide and Fab A were measured using a NanoDrop UV Spectrophotometer (Harlow Scientific). The level of binding was evaluated using fluorescence polarization (FP) by mixing equal volumes (20 μL) of the peptide and each Fab concentration in a 384 well black plate, resulting in a final peptide concentration of 250 nM. The dissociation constants (KD, μM) were determined. The first round of screen was performed with Fab A as target. Amplified phage from the first positive was diluted to a concentration of e11 pfu/200 μL and incubated thrice (first negative) in: (a) BSA blocked tube, for 1 hr at RT; (b, c) protein L magnetic beads, without Fab, for 50 and 40 min, respectively, at RT. In each incubation, the supernatant of the previous step was used. The supernatant after the first set of negative rounds was collected and incubated with Fab Z immobilized on Protein L magnetic beads, for 1 hr at RT (second positive). All the amplified phage from the second positive (concentration of e11 pfu/200 μL) was used in a second negative selection round, similar to the first negative (second negative). The supernatant of the second negative was collected and incubated with Fab D immobilized in protein L magnetic beads, for 45 min at RT. The amplified phage from the third positive was diluted to a concentration of e11 pfu/200 μL and added to four different tubes containing NHS beads (fourth positive) with: (a) a mixture of the three used Fabs, (b) Fab A, (c) Fab Z, and (d) Fab D. The top two peptide binders, A5 (WHYNWQDVSDRQ) and C7 (HQNHHSTFWEIY), were determined to have KD values of 4.1 μM and 6.5 μM, respectively. 3563 HAMRAQP(8)[243.3722±14.9784] NAPDWPA(4)[87.7108±23.5104] SPSTHWK(3)[24.0053±5.9724] STSFWIT(2)[41.7329±5.8776] NESHSRT(2)[26.1857±5.8776] GFFHKTT(1)[NT] LPAGRVL(1)[NT] NGLTAWT(1)[NT] AAPDWAG(1)[NT] HAKRARA(1)[NT] TLHPAAD(1)[NT] HVQLWAT(1)[NT] SAAIGTL(1)[NT] LSNNNLR(1)[NT] HAXRA,APDW 14 3 PMID:35103339 Colon cancer cell line SW480 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Binding selectivity of the frequently occurring selected phage clones to colon cancer cells was analyzed by cell-based ELISA. ELISA values for binding of each phage clone to each cell type were calculated by dividing the OD of selected phage clone to the OD of control phage (no displayed peptide). Data are reproduced from Figure 4 and presented as the mean ± SD. In the first round of in vitro selection, an aliquot of the primary library was added to an empty well (depletion well). The supernatant composed of phages that do not bind to the plastic surface of six-well plates was removed and poured into a serum-treated well. The supernatant containing phages that do not interact with serum was removed and transferred to the well with blocked absorber cells. The subtracted phage supernatant was recovered, applied to the target SW480 cells. According to the results of cell binding assay and phage cell-based ELISA, one of the isolated peptides denoted as CCBP1 (with the sequence HAMRAQP) was indicated to have the highest binding efficiency, selectivity, and specificity toward colon cancer cells with no significant binding to control cells. Peptide competitive inhibition assay revealed that binding of the phage-displayed CCBP1 is competitively inhibited by the same free peptide, suggesting that CCBP1 specific binding to the target cell is independent of the phage context. Taken together, our findings provide support for the notion that CCBP1 binds specifically to colon cancer cells and might be a potential lead candidate for targeted delivery of imaging agents or therapeutic genes/drugs to colon tumors. 3564 EHHRSHL(4)[0.6674±0.0648] HGSGVHA(3)[0.4796±0.076] APGGHSS(3)[0.8127±0.0425] VGYSGRD(2)[0.9651±0.0445] YMNDRMY(2)[0.8104±0.0516] DKSHVGL(2)[0.8398±0.0155] HPIKHLR(2)[0.7032±0.0335] SAQIAPH(2)[1.0009±0.0267] TGLIGQK(1)[0.5511±0.0335] GTQFFNK(1)[0.9448±0.0267] GYWNKFD(1)[0.7054±0.067] 12 Phage display (subtractive panning) 4 PMID:34202166 Copper ion, Cu(2+) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA An ELISA assay to determine the binding affinity of the 12 selected peptides to Cu(II) was performed. Absorbance was recorded at 405 nm with an ultraviolet spectrophotometer (UV 1800 Pharma Spec, Japan). Data are reproduced from Figure 2A and presented as the mean ± SE. In order to eliminate the phages bound to the resin, the IDA resin was first subjected to reverse biopanning followed by Cu(II) affinity biopanning. In brief, 100 μL of the phage solution (~2e12 plaque forming units, PFUS) in the NEB original peptide library was dissolved in 900 μL of TBST solution and added to 100 μL of IDA resin, followed by gently shaking for 25 min at 25°C. The supernatant was extracted and further infected with E. coli ER2738 cell culture for amplification according to the manufacturer’s instructions. Then affinity screening was carried out for screening Cu(II)-binding peptide. 100 μL of the phage solution (~0.8e12 virions) from the reverse screening amplification was dissolved in 900 μL of TBST solution, and added to 100 μL of Cu(II) chelating resin, followed by gently shaking for 25 min at 25°C. The supernatant was discarded, and the resin was washed twice with 1 mL of TBST solution to remove unbound phages. Finally, 1 mL of EDTA (0.5 M, pH 8.0) was added, and the eluate was collected by shaking at 200 rpm at 25°C for 10 min. The eluate was centrifuged with a 100 KD centrifugal filter device (Millipore, U.S.) at 4°C, 5000 rpm, 15 min to remove Cu(II). The remaining phages are amplified and subjected to the next round of screening. Besides, to obtain high affinity phages, 4 rounds of biopanning were conducted. In the fourth round of biopanning, Cu(II) loaded on the resin beads was reduced by washing with citrate buffer solution (pH 4.4) to improve the biopanning affinity. The Cu(II)-binding peptide (SAQIAPH, PCu) identified from the phage display heptapeptide library was used to explore the mechanism of PCu inhibition of Cu2+-mediated Aβ aggregation and Aβ production. In vitro experiments revealed that PCu directly inhibited Cu2+-mediated Aβ aggregation and regulated copper levels to reduce biological toxicity. Furthermore, PCu reduced the production of Aβ by inhibiting Cu2+-induced BACE1 expression and improving Cu(II)-mediated cell oxidative damage. Cell culture experiments further demonstrated that PCu had relatively low toxicity. This Cu(II)-binding peptide that we have identified using phage display technology provides a potential therapeutic approach to prevent or treat Alzheimer's disease. 3565 GWRVSEF(24) GWRVSEL(1) GFHYSLH(1) IVGSQVT(1) GWRVSE 4 3 PMID:34314166 Cowpea mosaic virus, CPMV Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The plate was incubated in the dark for 10 min, and the absorbance was measured at 370 nm by using an Infinite 200 Rx plate reader (Tecan Life Sciences) with 25 flashes in 96-well flat-bottom plate mode. The input library was preincubated overnight with blocking agent (2% (w/v) bovine serum albumin; BSA) each round to remove potential BSA binders. Biopanning and characterization of lead candidates resulted in isolation of the motif “GWRVSEF/L” as the plant virus cowpea mosaic virus (CPMV) specific motif with phenylalanine (F) at the seventh position being stronger than leucine (L). Specificity to CPMV was demonstrated, and cross-reactivity toward other plant viruses was not observed. To demonstrate cargo loading, GWRVSEF was tagged with biotin, fluorescein isothiocyanate (FITC), and a human epidermal growth factor receptor 2 (HER2)-specific targeting peptide ligand. Display of the active ingredient was confirmed, and utility of tagged and targeted CPMV in cell binding assays was demonstrated. The CPMV-binding peptides (CBP) functionalization strategy offers a new avenue for CPMV nanoparticle functionalization and should offer a versatile tool to add active ingredients that otherwise may be difficult to conjugate or display. 3566 AHWNPFWLATPF[0.409] YWVDSSAWVAHK[0.3389] NNDPLQLRSQRY[0.4388] KLDVFTKPLVFT[0.2597] 4 3 PMID:35755076 Purified serum IgG from human T-lymphotropic virus 1 (HTLV-1) infected people Human T-lymphotropic virus 1 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA After the screening performed in the first Phage ELISA, the phage clones containing the peptides that presented a ratio between the absorbances of the positive and control samples >2 were submitted to a second Phage ELISA. Although most of the peptides could discriminate individuals with HTLV-1 infection from seronegative control individuals, four peptides (A6, A8, B6, and D7) stood out from the rest. In the phage ELISA, dilutions of 1:50, 1:100, 1:500, and 1:1000 were used, and the best results were obtained at a dilution of 1:1000. The biopanning process for selection of HTLV-1 peptides included a subtractive step, which consisted of incubating the phage library (1.0e11 phage particles of the PD library) first with IgG from HTLV-seronegative individuals for 30 min at room temperature. Then, the supernatant containing unbound phages was added to IgG of HTLV-2-seropositive individuals with incubation for 30 min at room temperature. The next step consisted of positive selection, in which the final supernatant of the subtractive step was incubated with purified IgG from HTLV-1-positive individuals for 30 min at room temperature. Three biopanning cycles were performed for the selection of HTLV-1 peptides.The phages bound to the magnetic microspheres of the positive selections were recovered by acid elution and subjected to steps of amplification, titration, supernatant production, DNA extraction, and phage DNA sequencing. Bioprospection of peptides mimicking HTLV-1 using phage display led to the identification of four clones, three related to gp46 (A6, B6, and D7) and one related to protease and Tax (A8). The analysis of accessibility, antigenicity, and hydrophilicity showed that clones A6 and B6 are potentially suitable for diagnostic use and that clone B6 has great potential for use in vaccine tests. The best reactivity, evaluated by phage ELISA, was that of clone B6, which showed good AUC, sensitivity, specificity, and LR. Thus, the successful testing of the B6 clone peptide would be relevant to diagnostic tests on platforms that allow rapid results and to the availability of more affordable testing for the overall population, and it would contribute to the prevention and control of HTLV-1 infection. 3567 CTDKASSSC[0.6487±0.0543] CHMYHNATC[0.5415±0.0545] 2 Phage display (competitive panning) 4 PMID:35032803 Ovomucoid Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA ELISA signal was measured using a microplate spectrophotometer (Multiskan FC, Thermo Scientific, Waltham, MA, USA) at 405 nm. Streptavidin-coated plates (streptavidin high binding capacity coated 96-well plates) (Thermo Scientific, USA) were used for phage display. After adding 1 μL (1.0e11 PFU/mL) of each M13 library to 100 μL (24.75 μg/mL) of biotinylated ovomucoid, the mixture was stirred at 100 rpm for 1 h at room temperature. When the agitation of the phage-protein complex was completed, the streptavidin-coated plate was washed three times repeatedly using 0.1 M PBS, pH 7.2. Thereafter, the phage-protein complex was added to the streptavidin-coated plates and allowed to react at 110 rpm for 10 min to enable the specific binding between avidin and biotin. Then, 1 μL of biotin (0.1 mM) was added to the plate and incubated for 5 min for blocking, followed by repeated washing 10 times using PBST containing 0.1 M PBS with 0.1 % Tween 20. Finally, phages specifically bound to ovomucoid were eluted in 0.2 M Glycine-HCl (pH 2.2) 1 mg/mL BSA solution, and 15 μL of Tris-HCl (pH 9.0) was added for neutralization. After the characterization of binding affinities of selected phages, whole phage particles were covalently attached to a gold electrode using crosslinking chemistry (MUA-EDC/NHS and Sulfo-LC/SPDP); the developed phage sensor was characterized using cyclic voltammetry (CV), square wave voltammetry (SWV), and electrochemical impedance spectroscopy (EIS). The cyclic peptide (CTDKASSSC) displayed phage sensor modified using EDC/NHS chemistry exhibited significantly better binding affinity (Kd = 2.36 ± 0.44 μg/mL) and limit of detection (LOD, 0.12 μg/mL) for ovomucoid than the linear phage sensor, resulting in good reproducibility and recovery, even in an actual egg and white wine samples. This approach may provide an alternative and more efficient way of sensing food allergens with desirable sensitivity, selectivity, and feasibility in food diagnostic applications. 3568 LQAYIGPKATWW[0.5068±0.0774] HHSRFSTLFNWP[0.5013±0.0917] WWQPYSSAPRWL[0.8965±0.0631] 3 Phage display (competitive panning) 4 PMID:35032803 Ovomucoid Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA ELISA measurements were performed to compare the relative binding affinities of the five M13 phage candidates as follows. First, the streptavidin-coated plate was pre-washed thrice with 200 μL 0.1 M PBS for 5 min each, following which 100 μL biotinylated ovomucoid (24.75 μg/mL) was added to the streptavidin-coated plate and stirred at room temperature for 1 h. Subsequently, the protein solution was removed and 200 μL blocking solution (5% BSA in NaHCO3, pH 8.6) was added and incubated at 4 ℃ for 1.5 h. Then, after repeated washing with 0.5% PBST six times, each phage was added at a concentration of 1012 PFU/mL, followed by stirring at 100 rpm at room temperature for 1 h. After washing six times with 0.5 % PBST, horseradish peroxidase (HRP)-conjugated anti-M13 monoclonal antibody (1:2,500 v/v) diluted in blocking solution was added and the microplate was then incubated for 1 h at 25 °C. Unbound antibody solution was removed, and the microplate was washed again with TBST. Freshly prepared HRP substrates, tetramethylbenzidine (TMB) were introduced to the microplate and the ELISA signal was measured using a microplate spectrophotometer (Multiskan FC, Thermo Scientific, Waltham, MA, USA) at 405 nm. Streptavidin-coated plates (streptavidin high binding capacity coated 96-well plates) (Thermo Scientific, USA) were used for phage display. After adding 1 μL (1.0e11 PFU/mL) of each M13 library to 100 μL (24.75 μg/mL) of biotinylated ovomucoid, the mixture was stirred at 100 rpm for 1 h at room temperature. When the agitation of the phage-protein complex was completed, the streptavidin-coated plate was washed three times repeatedly using 0.1 M PBS, pH 7.2. Thereafter, the phage-protein complex was added to the streptavidin-coated plates and allowed to react at 110 rpm for 10 min to enable the specific binding between avidin and biotin. Then, 1 μL of biotin (0.1 mM) was added to the plate and incubated for 5 min for blocking, followed by repeated washing 10 times using PBST containing 0.1 M PBS with 0.1 % Tween 20. Finally, phages specifically bound to ovomucoid were eluted in 0.2 M Glycine-HCl (pH 2.2) 1 mg/mL BSA solution, and 15 μL of Tris-HCl (pH 9.0) was added for neutralization. 3569 IPPVPTTTKASV(4)[48.1236±3.7248] WHWTWLSEYPPP(3)[34.79±5.3442] HWTSFSWLGSWN(3)[55.1952±1.9973] IPPVPRLKTSYK(2)[69.8783±7.0177] FHWSLPWLPGLP(2)[77.5438±3.3469] WHFEWWRATPSG(2)[31.767±5.0203] HWWNTPWWAWHP(2)[43.1572±7.9893] HWNFWMHNAHWN(2)[46.828±1.9973] HTWWLWPPQLPP(2)[54.4935±3.6708] WHTTELLSLPWR(1)[31.1192±13.0097] WHWNPFRVSQPL(1)[32.4148±1.0257] WHFSYSYKPALS(1)[36.7874±12.038] HWNFWMQNATRS(1)[46.828±2.3212] ILLGRRLKYSTA(1)[47.4758±3.6708] WHTTELLSLPWR(1)[48.1236±3.7248] WHWNAWALPTHG(1)[54.4935±5.3442] NHWPSYFAQWMT(1)[89.2579±0.7018] WHW, HW, WH and HWW 17 4 PMID:35068948 72 kDa type IV collagenase Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) MMP-2 (15 ng) was incubated overnight with 2e11 pfu/mL phage clones that were enriched for binding against active MMP-2. After 18 h, each mixture was run on 7.5% SDS-PAGE with 20 mg/mL gelatin. After electrophoresis, the gel was washed twice at room temperature with 2.5% Triton-X 100 for 30 min. The gel was then kept overnight in zymogram retention solution (6.06 g Tris-HCl, 1.47 g CaCl2, and 2.92 g NaCl per liter). The next day, the gel was stained with Coomassie Blue R. The density of the bands were analyzed using the Bio-Rad Multi-Analyst program (Tajhya et al., 2017). The gel was scanned using a digital scanner, and MMP-2 activity was determined by measuring the peak area of each band. The peak percentage was normalized relative to the activity of MMP-2 (positive control) and wild-type M13 phage (negative control). The inhibitory effects of the peptides on the gelatinase activity of MMP-2 was reproduced from Figure 4. For subtractive screening, 200 μL of the Ph.D.-12 library (containing 4e10 phages) was first added to wells coated with active matrix metalloproteinase-9 (MMP-9) ligand incubated for 1 h at room temperature. Unbound phages were recovered and transferred to wells coated with active-MMP-2. MMP-2 inhibition by selected peptides was evaluated through zymogram analyses, which revealed that four peptides (WHFEWWRATPSG, WHWNPFRVSOPL, WHTTELLSLPWR and WHWTWLSEYPPP) inhibited MMP-2 activity by at least 65%. These four peptides were synthesized and used for in vitro wound healing using human umbilical vein endothelial cells, and two peptides, AOMP12 (WHTTELLSLPWR) and AOMP29 (WHWTWLSEYPPP), were found to inhibit wound healing by 40%. These peptides are, thus, potential candidates for MMP- 2 inhibition for cancer treatment. Furthermore, our findings suggest that our substractive biopanning screening method is a suitable strategy for identifying peptides that selectively inhibit MMP-2. 3570 SLNTTWVSPMMK(19) YNTHGVRDNAWL(6) IESRYLTKEAVH(3) 3 Phage display (common panning) 2-4 PMID:37044252 Microcentrifuge tube (polypropylene, PP) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3571 TPPSSNSYDWLV(17) METRPVAPHEFR(8) ALKIGPETTIYM(4) 3 Phage display (common panning) 2-4 PMID:37044252 Boiled Daphnia magna asexual egg Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) 3572 LYALPLSHLKSH(13)[95.55%] SPSSAYPRHGPD(12)[87.50%] DPLLFPGTSRQM(6)[47.00%] 3 Phage display (common panning) 2-4 PMID:37044252 Daphnia magna asexual egg Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay Ten microliters of each phage was added to 200 μL of ER2738 culture grown to an OD600 of 0.7 and incubated at RT for 5 min. The cells with infected phages were transferred to a conical tube containing 5 mL of Top Agar at around 40 ◦C and poured directly onto an LB/IPTG/Xgal plate containing 1 mL of IPTG/Xgal stock in 25 mL of DMF. After standing for 1 h to solidify, the plates were incubated at 37 ◦C overnight. After counting the number of blue plaques on each plate, we calculated the pfu/mL value while considering the dilution factor of each phage. Attached % was calculated as: ((Input phage-Output phage)/Input phage) × 100. We identified a peptide, DEP1 (LYALPLSHLKSH), with the highest binding affinity to D. magna eggs. DEP1 did not affect zebrafish eggs, but it inhibited normal hatching and reproductive ability in D. magna eggs, and hindered growth in neonates before their first ecdysis. Morphological analysis revealed that DEP1 caused intestinal damage and tissue abnormalities. Our findings demonstrate that the whole cell-based phage display technique is successful in presenting antigens in their natural form, and that the DEP1 peptide can be applied to regulate the growth cycle of D. magna. These results have implications for the use of phage display in environmental research and the potential use of DEP1 for hazardous organisms in aquatic systems. 3573 CTEWDYLTVC 1 Phage display (common panning) 4 PMID:33508757 Prostate specific antigen (PSA(-/lo)) human prostate cancer cell line LNCaP Not determined. Not determined. Not determined. fUSE55-based CX8C phage display library (CX8C) Prostate cancer (PCa) contains both differentiated (PSA(+)) and undifferentiated (PSA(−/lo)) tumor cells. PSA was short for prostate-specific antigen. PSA(+) and PSA(-/lo) cells were mixed in equal proportions and incubated with CX8C phage display library. A measure of 1.0e9 of the phage display peptide library in 5 ml of 1 ×phosphate buffered saline (PBS) was added to the confluent cell monolayer and incubated for 2 h on a rocker platform at 4 °C. The cells were washed four times with PBS containing 0.2% Tween 20. Cells were subjected to fluorescence-activated cell sorting (FACS) for GFP(+) and GFP(−) cells. Sorted cells were lysed to release phage particles that were used to infect K91 bacteria. Individual bacterial colonies that emerged on the Tet/Kanamycin plates were isolated, amplified, and phage particles reisolated and used in 2–3 more similar cycles. PSA(−/lo) PCa cell-specific targeting peptide (TAP1, CTEWDYLTVC) suppressed PCa cell growth both in vitro and in vivo and improved the drug sensitivities of anti-androgens and chemotherapeutic agents at least through shortening the length of telomere and reducing the expression of HOXB9 and TGF-β2. 3574 STTGTQY(4)[30.7011±2.0436] DLFVSSL(1)[NT] VNLNLLP(1)[NT] SIWQSLN(1)[NT] AKLHILR(1)[NT] KHINPSI(1)[NT] YWPGYSM(1)[NT] SPLLIPQ(1)[NT] NQVLARH(1)[NT] TWQVLRP(1)[NT] YDRENHP(1)[NT] LEVTPWW(1)[NT] YLSPVPM(1)[NT] 13 Phage display (subtractive panning) 3 PMID:33706101 Human embryonic kidney cells 293 (HEK293 cells) transfected with free fatty acid receptor 1 (FFAR1) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) HEK293 cells were seeded in a 24-well plate at a density of 1.0e5 cells/well and incubated overnight. Expression vectors (pCAGGS/FFAR1 and pCAGGS/APTGF- a) were kindly provided by Dr. Inoue (Tohoku University) and used for transfection. A mixture of 125 ng pCAGGS/AP-TGF-α and 50 ng pCAGGS/FFAR1 per well in 24-well plates was transfected using PEI (49553-93-7, Polysciences, Warrington, PA, USA) and incubated overnight. The following day, HEK293 cells were harvested by trypsinization and re-seeded in 96-well plates at a volume of 80 mL/well. After incubation for 1 h, 20 mL of palmitic acid (PA) and peptide samples in HBSS containing 5 mM HEPES (pH 7.4) were added and incubated for 1 h. Eighty microliters of supernatant containing released AP-TGF-α was transferred into an empty 96- well plate. A substrate for alkaline phosphatase, paranitrophenylphosphate (p-NPP, 34045, Thermo Fisher Scientific) was added to the transferred medium, and absorbance at 405 nm of the plates was measured using a microplate reader before and after 30 min incubation at 37 ◦C. We calculated AP-TGF-α release as a TGF-α shedding response. First, 1.0e7 HEK293 cells in 1 mL phosphate buffered saline (PBS) containing 1% bovine serum albumin (BSA) were incubated with 1.5e11 phages on a shaking machine (120 rpm) for 2 h at 4 °C. This mixture was centrifuged at 1500 rpm for 1 min, and the supernatant containing phages that could not bind HEK293 cells was incubated with other HEK293 cells. This selection (negative selection) was repeated twice to remove phage-binding HEK293 cells. Next, the supernatant was incubated with 5.0e6 HEK293/FFAR1 cells on a shaking machine (120 rpm) for 2 h at 4 °C. The HEK293/FFAR1 cells were washed five times in cold PBS containing 1% BSA and 0.1% Tween-20. HEK293/FFAR1 cells binding phages were eluted with Glycine-HCl (pH 9.1) containing 1 mg/mL BSA on ice for 10 min. The 10 μL phages were titered according to the manufacturer’s instructions, and the amplified phage using E. coli ER2738 was used in the next round of selection. STTGTQY determined by phage display promoted glucose-stimulated insulin secretion in pancreatic MIN6 cells. 3575 DLFVSSL(18)[24.3461±1.1215] AETVESC(2)[NT] STTGTQY(1)[NT] WSLDPSS(1)[NT] SLVVERL(1)[NT] 5 Phage display (subtractive panning) 4 PMID:33706101 Human embryonic kidney cells 293 (HEK293 cells) transfected with free fatty acid receptor 1 (FFAR1) Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) HEK293 cells were seeded in a 24-well plate at a density of 1.0e5 cells/well and incubated overnight. Expression vectors (pCAGGS/FFAR1 and pCAGGS/APTGF- a) were kindly provided by Dr. Inoue (Tohoku University) and used for transfection. A mixture of 125 ng pCAGGS/AP-TGF-α and 50 ng pCAGGS/FFAR1 per well in 24-well plates was transfected using PEI (49553-93-7, Polysciences, Warrington, PA, USA) and incubated overnight. The following day, HEK293 cells were harvested by trypsinization and re-seeded in 96-well plates at a volume of 80 mL/well. After incubation for 1 h, 20 mL of palmitic acid (PA) and peptide samples in HBSS containing 5 mM HEPES (pH 7.4) were added and incubated for 1 h. Eighty microliters of supernatant containing released AP-TGF-α was transferred into an empty 96- well plate. A substrate for alkaline phosphatase, paranitrophenylphosphate (p-NPP, 34045, Thermo Fisher Scientific) was added to the transferred medium, and absorbance at 405 nm of the plates was measured using a microplate reader before and after 30 min incubation at 37 ◦C. We calculated AP-TGF-α release as a TGF-α shedding response. First, 1.0e7 HEK293 cells in 1 mL phosphate buffered saline (PBS) containing 1% bovine serum albumin (BSA) were incubated with 1.5e11 phages on a shaking machine (120 rpm) for 2 h at 4 °C. This mixture was centrifuged at 1500 rpm for 1 min, and the supernatant containing phages that could not bind HEK293 cells was incubated with other HEK293 cells. This selection (negative selection) was repeated twice to remove phage-binding HEK293 cells. Next, the supernatant was incubated with 5.0e6 HEK293/FFAR1 cells on a shaking machine (120 rpm) for 2 h at 4 °C. The HEK293/FFAR1 cells were washed five times in cold PBS containing 1% BSA and 0.1% Tween-20. HEK293/FFAR1 cells binding phages were eluted with Glycine-HCl (pH 9.1) containing 1 mg/mL BSA on ice for 10 min. The 10 μL phages were titered according to the manufacturer’s instructions, and the amplified phage using E. coli ER2738 was used in the next round of selection. DLFVSSL showed free fatty acid receptor 1 (FFAR1) agonist activity, although it was lower than that of STTGTQY. 3576 WDMWPSMDWKAE(4/27)[1.0403±0.0736] FGLEPRANLHFT(2/27)[0.6586±0.0164] VQVRDNLPTTTG(2/27)[0.2498±0.0172] THDMGKMDRTTT(2/27)[0.307±0.0265] WGNSQWTGQHTT(1/27)[0.7087±0.0236] 5 Phage display (subtractive panning) 2-3 PMID:35168735 Interleukin-33 (IL-33) Interleukin-1 receptor-like 1 Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA ELISA was performed to evaluate the binding affinity of the phage particles to IL-33. First, streptavidin-coated wells were incubated with 100 μL of biotinylated IL-33 (1 μM) at room temperature for 1 h. The supernatant with residual proteins or unbound phages was subsequently discarded, and then, 200 μL of blocking buffer comprising 0.1 M NaHCO3 (pH 8.6) with 5 mg/mL of BSA and 0.02% NaN3, was added to the wells and incubated for 1 h at 4 °C. The wells were filled with blocking buffer. After incubation, the plate wells were washed 6 times with 0.5% PBST (0.1 M PBS (pH 7.4) with 0.5% Tween 20). The screened peptide-displaying phages (1 × 1011 or 1 × 1012 pfu/mL) were added to the pre-functionalized wells and incubated for 1 h at room temperature with mild shaking. The phage solutions were removed, and then the wells were washed six times with 0.5% PBST to remove unbound phages. The wells were then filled with HRP-conjugated anti-M13 monoclonal antibodies (diluted 1:5000 in blocking buffer) and incubated for 1 h at room temperature with mild shaking. Finally, the wells were washed with 0.5% PBST and filled with 200 μL ABTS (with H2O2), which is an HRP substrate. The change of absorbance was measured at 405 nm using a microplate spectrophotometer (Multiskan FC, Thermo Scientific, Waltham, MA, USA). The change of absorbance at 405 nm was reproduced from Figure 1A. We performed three rounds of biopanning against IL-33 protein. A negative selection beginning with the 2nd round, in which the amplified phage was preincubated with the streptavidin-coated plate in the absence of IL-33, was performed. The selected IL-33 specific peptide was identified as FGLEPRANLHFT. To investigate the molecular interactions between IL-33 and the affinity peptide, the peptide was separated from the phage particles, chemically synthesized and characterized by square wave voltammetry (SWV), isothermal titration calorimetry (ITC), and microscale thermophoresis (MST). The binding constant (Kd) value with SWV, MST, and ITC was found to be 1.68 ± 0.37 μM, 5.98 ± 1.30 μM, and 2.68 ± 1.37 μM, respectively. Two-dimensional (2D) NMR spectral analysis was performed to elucidate the primary peptide binding site of IL-33, which was near the ST2-D3 and IL1RAcP-D3 binding interfaces. 3577 CPMKSHTNC(4) CQGTSLSHC(1) CDQRLPNYC(1) CITNHSPTC(1) CPTGPSATC(1) CTSSEPNLC(1) CNSSRSELC(1) 7 Phage display (in vivo) 3 PMID:34185171 Cerebrospinal fluid Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) To obtain the targeting peptides, the appropriate time to recover phages from the CSF was determined using a recovery time curve. The titer (in transducing units/ml CSF) of the phages recovered was plotted against time to construct the recovery time curve (Li et al. 2015). Adult male Sprague–Dawley rats were used for three rounds of screening. In the first round, Sprague–Dawley rats (n = 3) were injected intravenously (i.v.) with 1012 pfu of Ph.D.-C7C™ phage display library suspended in 100 μl of tris-buffered saline (TBS) (50 mM Tris–HCl, 150 mM NaCl, pH 7.5). The phages were allowed to circulate in the rats for 1 h before CSF was drawn. Sprague–Dawley rats were anesthetized using 5% chloral hydrate (0.4 g/kg). Phages recovered from the CSF were then incubated with E. coli (ER2738 host strain) for amplification. The other two rounds of biopanning were proceeded by injecting rats (i.v.) with newly amplified phages (1 × 1012 pfu in 100 μl TBS) and repeating the subsequent procedures of the first round as described above. A peptide sequence denoted as PMK (CPMKSHTNC), which was demonstrated to be able to cross the blood-cerebrospinal fluid barrier (BCSFB) via in vivo optical imaging analysis, could be used in the future for the construction of targeted drug delivery systems. 3578 DYHDPSLPTLRK(30.2%)[0.0321±0.0062] QVNGLGERSQQM(15.7%)[NT] RDYHPRDHTATW(7.9%)[NT] GNNPLHVHHDKR(4.0%)[NT] TAKYLPMRPGPL(3.3%)[NT] SPLRAVAFSGAQ(2.4%)[NT] INIVPGPEKPVG(2.2%)[NT] QGYKQEYTRWGE(<0.1%)[NT] 8 Phage display (common panning) 1 PMID:36861429 Surfaces of crystalline gypsum Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The affinity of selected oligopeptides and synthesized copolymers to bind to the surface of gypsum and C-S-H particles was assessed in batch assays based on total organic carbon (TOC) analysis. For the selection phase, the following procedure was used: 10 µL of the original phage library solution was mixed with 990 µL saturated gypsum solution (to prevent the dissolution of the gypsum particles during the experiment) in standard Eppendorf vials. To this dilution, solid gypsum powder was added at a ratio of 0.1% (w/v) and the resulting suspension was incubated at room temperature for 30 min while shaking at 800 rpm. Afterward, the solid particles carrying the strongly adhering phages were separated by centrifugation for 30 s at 8000 rpm, while the non- and weakly bound phages remained in the supernatant, which was removed. The remaining solid material was washed ten times with 1 mL saturated gypsum solution by vortexing for 15 s and subsequent centrifugation, in order to further separate weakly from strongly binding phages. During the washing sequence, the Eppendorf vial was replaced three times to eliminate any potential influence of phages sticking to the polypropylene surface of the vials (i.e., to increase the selection pressure and remove peptide sequences with enhanced affinity for the vial surfaces prior to analysis). To isolate the strongly bound phages in the last step, the washed gypsum particles were dissolved in a mixture of 1 mL Tris/HCl and 100 µL Tris-buffered saline. Subsequently, the released phages were amplified in a 20 mL lysogeny broth (LB) medium containing 400 µL Escherichia Coli solution (strain ER2738, OD600 ≈ 0.5). After 4-5 hours of vigorous shaking at 37 °C, the phages were reprocessed according to the protocol of the supplier to prepare the selected library for another panning round. Based on next-generation sequencing of phages enriched during the screening process, a triplet of amino acids, DYH, is identified as the main driver for adsorption on the mineral substrate. Furthermore, oligopeptides containing this motif prove to exert their influence in a strictly selective manner during the hydration of cement, where the sulfate reaction (initial setting) is strongly retarded while the silicate reaction (final hardening) remains unaffected. In the final step, these desired additive characteristics are successfully translated from the level of peptides to that of scalable synthetic copolymers. 3579 DYHDPSLPTLRK(76.0%)[0.0321±0.0062] TAKYLPMRPGPL(7.9%)[NT] QVNGLGERSQQM(4.3%)[NT] RDYHPRDHTATW(2.6%)[NT] GNNPLHVHHDKR(1.2%)[NT] SPLRAVAFSGAQ(0.6%)[NT] INIVPGPEKPVG(0.5%)[NT] QGYKQEYTRWGE(0.1%)[NT] 8 Phage display (common panning) 2 PMID:36861429 Surfaces of crystalline gypsum Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The affinity of selected oligopeptides and synthesized copolymers to bind to the surface of gypsum and C-S-H particles was assessed in batch assays based on total organic carbon (TOC) analysis. For the selection phase, the following procedure was used: 10 µL of the original phage library solution was mixed with 990 µL saturated gypsum solution (to prevent the dissolution of the gypsum particles during the experiment) in standard Eppendorf vials. To this dilution, solid gypsum powder was added at a ratio of 0.1% (w/v) and the resulting suspension was incubated at room temperature for 30 min while shaking at 800 rpm. Afterward, the solid particles carrying the strongly adhering phages were separated by centrifugation for 30 s at 8000 rpm, while the non- and weakly bound phages remained in the supernatant, which was removed. The remaining solid material was washed ten times with 1 mL saturated gypsum solution by vortexing for 15 s and subsequent centrifugation, in order to further separate weakly from strongly binding phages. During the washing sequence, the Eppendorf vial was replaced three times to eliminate any potential influence of phages sticking to the polypropylene surface of the vials (i.e., to increase the selection pressure and remove peptide sequences with enhanced affinity for the vial surfaces prior to analysis). To isolate the strongly bound phages in the last step, the washed gypsum particles were dissolved in a mixture of 1 mL Tris/HCl and 100 µL Tris-buffered saline. Subsequently, the released phages were amplified in a 20 mL lysogeny broth (LB) medium containing 400 µL Escherichia Coli solution (strain ER2738, OD600 ≈ 0.5). After 4-5 hours of vigorous shaking at 37 °C, the phages were reprocessed according to the protocol of the supplier to prepare the selected library for another panning round. Based on next-generation sequencing of phages enriched during the screening process, a triplet of amino acids, DYH, is identified as the main driver for adsorption on the mineral substrate. Furthermore, oligopeptides containing this motif prove to exert their influence in a strictly selective manner during the hydration of cement, where the sulfate reaction (initial setting) is strongly retarded while the silicate reaction (final hardening) remains unaffected. In the final step, these desired additive characteristics are successfully translated from the level of peptides to that of scalable synthetic copolymers. 3580 DYHDPSLPTLRK(86.3%)[0.0321±0.0062] TAKYLPMRPGPL(7.8%)[NT] QVNGLGERSQQM(0.9%)[NT] RDYHPRDHTATW(0.7%)[NT] GNNPLHVHHDKR(0.4%)[NT] SPLRAVAFSGAQ(0.1%)[NT] INIVPGPEKPVG(0.1%)[NT] QGYKQEYTRWGE(0.1%)[NT] 8 Phage display (common panning) 3 PMID:36861429 Surfaces of crystalline gypsum Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The affinity of selected oligopeptides and synthesized copolymers to bind to the surface of gypsum and C-S-H particles was assessed in batch assays based on total organic carbon (TOC) analysis. For the selection phase, the following procedure was used: 10 µL of the original phage library solution was mixed with 990 µL saturated gypsum solution (to prevent the dissolution of the gypsum particles during the experiment) in standard Eppendorf vials. To this dilution, solid gypsum powder was added at a ratio of 0.1% (w/v) and the resulting suspension was incubated at room temperature for 30 min while shaking at 800 rpm. Afterward, the solid particles carrying the strongly adhering phages were separated by centrifugation for 30 s at 8000 rpm, while the non- and weakly bound phages remained in the supernatant, which was removed. The remaining solid material was washed ten times with 1 mL saturated gypsum solution by vortexing for 15 s and subsequent centrifugation, in order to further separate weakly from strongly binding phages. During the washing sequence, the Eppendorf vial was replaced three times to eliminate any potential influence of phages sticking to the polypropylene surface of the vials (i.e., to increase the selection pressure and remove peptide sequences with enhanced affinity for the vial surfaces prior to analysis). To isolate the strongly bound phages in the last step, the washed gypsum particles were dissolved in a mixture of 1 mL Tris/HCl and 100 µL Tris-buffered saline. Subsequently, the released phages were amplified in a 20 mL lysogeny broth (LB) medium containing 400 µL Escherichia Coli solution (strain ER2738, OD600 ≈ 0.5). After 4-5 hours of vigorous shaking at 37 °C, the phages were reprocessed according to the protocol of the supplier to prepare the selected library for another panning round. Based on next-generation sequencing of phages enriched during the screening process, a triplet of amino acids, DYH, is identified as the main driver for adsorption on the mineral substrate. Furthermore, oligopeptides containing this motif prove to exert their influence in a strictly selective manner during the hydration of cement, where the sulfate reaction (initial setting) is strongly retarded while the silicate reaction (final hardening) remains unaffected. In the final step, these desired additive characteristics are successfully translated from the level of peptides to that of scalable synthetic copolymers. 3581 VIELVILIDD APELLHLIDE HKELSQLIWF PKELNRLIFG YGELGYLIHG EHELTLLILF YMELFILIAI SIELRHLIYE GDELHKLILY FWELNILIVY 10 Phage display (common panning) 3 PMID:35580187 Amyloid β peptide (Aβ) (M1-40) monomer, Aβ40 monomer Not determined. Not determined. Not determined. pIT2 X2ELX2LIX2 phage display library 3582 DQELEGLIHP SLELFPLIDD DAELNKLINP AYELPYLIIP IWELNILIEL AIELYALIFK DYELPNLIPP EFELGPLIEA GNELMQLIMM YYELDDLIMP 10 Phage display (common panning) 3 PMID:35580187 Amyloid β peptide (Aβ) (M1-40) fibril, Aβ40 fibril Not determined. Not determined. Not determined. pIT2 X2ELX2LIX2 phage display library 3583 TAELRNLIWA THELPILILS MAELHALIGY MRELVELIHV NEELEHLINE GPELYALIIF HNELPDLIQL HSELELLIEY VMELANLIGT HFELSRLINY 10 Phage display (common panning) 3 PMID:35580187 Amyloid β peptide (Aβ) (M1-42) monomer, Aβ42 monomer Not determined. Not determined. Not determined. pIT2 X2ELX2LIX2 phage display library 3584 WTELTVLIVW PTELQILIHW TPELAILISY PPELQNLIHF FLELDVLIHA YYELTELIYV RQELGFLILF KNELYWLIVE DSELKYLIYW DDELFMLIRM 10 Phage display (common panning) 3 PMID:35580187 Amyloid β peptide (Aβ) (M1-42) fibril, Aβ42 fibril Not determined. Not determined. Not determined. pIT2 X2ELX2LIX2 phage display library 3585 GGGVNIGLEYGGG GGGLFVMTRMGGG GGGHHYTVFMGGG GGGILALFFVGGG GGGEDHREMDGGG GGGHPRSTAVGGG GGGIMGYPLNGGG GGGFGVHEWVGGG GGGPTDIWAWGGG GGGPIHESEHGGG 10 Phage display (common panning) 3 PMID:35580187 Amyloid β peptide (Aβ) (M1-40) monomer, Aβ40 monomer STAT3 Y705 specific antibody Not determined. Not determined. pIT2 GGGX7GGG phage display library 3586 GGGIRQDAQAGGG GGGRHRKPFEGGG GGGGLDTRHDGGG GGGTLGKMHHGGG GGGKNMQMWVGGG GGGDYQPQGIGGG GGGWPVGHATGGG GGGMKQGPVYGGG GGGFKRSWIFGGG GGGHITHNETGGG 10 Phage display (common panning) 3 PMID:35580187 Amyloid β peptide (Aβ) (M1-40) fibril, Aβ40 fibril Not determined. Not determined. Not determined. pIT2 GGGX7GGG phage display library 3587 GGGQGKSVPAGGG GGGYLTIRLMGGG GGGASNTYFSGGG GGGLIWGFKTGGG GGGPDPLDFDGGG GGGDLVSFYYGGG GGGSWMALLVGGG GGGSSWRGTTGGG GGGHHQMTKSGGG GGGPWTVPVDGGG 10 Phage display (common panning) 3 PMID:35580187 Amyloid β peptide (Aβ) (M1-42) monomer, Aβ42 monomer Not determined. Not determined. Not determined. pIT2 GGGX7GGG phage display library We identified an SXkmer with loop–insertion YLTIRLM as an inhibitor of the secondary nucleation of Aβ42 and binding analyses using surface plasmon resonance technology, F€orster resonance energy transfer, and microfluidics diffusional sizing imply an interaction with intermediate oligomeric species. A linear peptide with the YLTIRLM sequence was found inhibitory but at a lower potency than the more constrained SXkmer loop. We identified an SXkmer with side-patch VI-WI-DD as an inhibitor of Aβ40 aggregation. Remarkably, our data imply that SXkmer-YLTIRLM blocks secondary nucleation through an interaction with oligomeric intermediates in solution or at the fibril surface, which is a unique inhibitory mechanism for a library-derived inhibitor. 3588 GGGEGVNEFFGGG GGGIRWTVMMGGG GGGGYRWWWVGGG GGGDRSNSPEGGG GGGRAHDASIGGG GGGVHTKAAAGGG GGGDQWIEHVGGG GGGNEMFVVWGGG GGGDRQWYPAGGG GGGVHKFGHIGGG 10 Phage display (common panning) 3 PMID:35580187 Amyloid β peptide (Aβ) (M1-42) fibril, Aβ42 fibril Not determined. Not determined. Not determined. pIT2 GGGX7GGG phage display library 3589 CPTTMWRYC(13)[1.4229±0.2161] CSTPMWKYC(5)[NT] CEKMVATHC(2)[NT] CTPRSANYC(2)[NT] CDGANARMC(1)[NT] CDGAPDAAC(1)[NT] CDQSVPHSC(1)[NT] CFGYYGPPC(1)[NT] CGSLDWPHC(1)[NT] CGTTEWRYC(1)[NT] CHNTYHRLC(1)[NT] CIVEASVHC(1)[NT] CMSWSNYFC(1)[NT] CNSSKLHMC(1)[NT] CPFWPSGHC(1)[NT] CPPMGOGNC(1)[NT] CPTTMWKHC(1)[NT] CQASRPALC(1)[NT] CQSCYCNHC(1)[NT] CQSYEPLRC(1)[NT] CRSTPWPSC(1)[NT] CSPHLNTNC(1)[NT] CSPWSYLFC(1)[NT] CSRSMDSTC(1)[NT] CSTGNNFAC(1)[NT] CSTLHQKLC(1)[NT] CTDKASSSC(1)[NT] CTKPNAPMC(1)[NT] CTNSGTSGC(1)[NT] CTSPMWRYC(1)[NT] CTTDVTGRC(1)[NT] GINNLPKSC(1)[NT] 32 Phage display (competitive panning) 2-4 PMID:35240465 Caspase-3, CASP-3 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA First, the streptavidin-coated 96-well plates were pre-washed three times with 200 μL of 0.1 M PBS before protein immobilization, and the biotinylated caspase-3 was added to the functionalized streptavidin-coated plate and stirred at 110 rpm for 1 h. Subsequently, the unbound proteins were removed and then added blocking solution (5% BSA in NaHCO3, pH 8.6) at 4 ℃ for 1.5 h. Then, the wells were washed six times with PBST buffer, and each phage were added at a concentration of 1012 PFU/mL, followed by stirring at 110 rpm at room temperature for 1 h. After washing six times with same buffer, horseradish peroxidase (HRP)-conjugated anti-M13 monoclonal antibody (1:2500 dilution in blocking buffer) was added and incubated for 1 h. The remaining solution was removed, and the plate was washed again with the same buffer. The ABTS and HRP substrate were added, and absorbance was measured at 405 nm (Multiskan, Thermo Scientific, Waltham, MA, USA). Absorbance at 405 nm was reproduced from Figure 1A. In the first round of biopanning, the biotinylated caspase-3 was separately pre-reacted with the Ph.D.-C7C random phage libraries (∼e11 PFU/mL) at 100 rpm for 1 h at room temperature. After mild agitation of the phage-protein complexes, 100 μL of the complexes was introduced to the streptavidin-coated 96-well plates (Thermo Scientific) and allowed to react at 110 rpm for 10 min to facilitate specific binding between avidin and biotin. Then, 1 μL of biotin (0.1 mM) was added to competitively remove the phages which is non-specifically binding to the streptavidin-coated plate as a blocking agent and incubated for 5 min, followed by repeated washing 10 times using PBST buffer containing 0.1 M PBS with 0.1% Tween 20 for removing unbound phages and residual biotin. Finally, the bound phages were eluted in 0.2 M glycine-HCl (pH 2.2) with 1 mg/mL BSA solution. The eluent was neutralized with 15 μL of Tris-HCl (pH 9.0) to prevent deactivation or destruction of the desired phages. The eluted phages were amplified using Escherichia coli ER2738, and the amplified phage solution was quantified by phage titration for subsequent rounds of biopanning. During this biopanning, negative biopanning against streptavidin-coated plate was performed with the amplified phages in the beginning of the second round, and Tween 20 concentration of PBST was gradually increased (0.1–0.5%) for removing non-specifically binding phages. We identified potential affinity peptide-displayed phage clones with the sequence CPTTMWRYC. After characterization of its binding affinity using enzyme-linked immunosorbent assay, whole phage particles were covalently attached to a gold surface using coupling chemistry (MUA-EDC/NHS). The developed phage sensor was characterized by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), scanning electron microscopy (SEM), electrochemical analysis using cyclic voltammetry (CV), and square wave voltammetry (SWV). Under optimal conditions, the affinity peptide-displayed phage sensor showed a good binding affinity (Kd = 0.13 ± 0.56 μM) and limit of detection (0.39 μM) for caspase-3 detection. Furthermore, developed phage sensor could be monitored the response of apoptotic HeLa cells by detecting caspase-3 activity. This work should stimulate the development of efficient alternative caspase-3 detection methods for the diagnosis and prognosis of apoptosis-related diseases. 3590 NWYLPWLGTNDW[++++] TWDLPWLLEKPF[++++] KMLPTMPRVLAG[++] DAAPTLPKGGVG[++] QIDTGYGLVSVS[++] GSKTGYLSETVR[++] ASKNAHLFLSSL[+] QQQYGTYVPTFG[+] 8 Phage display (subtractive panning) 3 PMID:37046671 M2 macrophages Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Flow Cytometry Binding of the phage clones to M2 macrophages was analyzed by flow cytometry. +, binding < 25%; ++, binding < 50%; ++++, binding > 80–100%. For the NW peptide (NWYLPWLGTNDW) blocking strategy, the M2 macrophages were incubated with 200 μg of the NW peptide for 1 h at RT prior to biopanning as described above. Three rounds of biopanning were performed on blocked M2 macrophages. Similarly, the library was pre-incubated with peripheral blood mononuclear cells (PBMCs) prior to affinity selection on blocked M2 macrophages. 3591 NWYLPWLGTNDW[++++] QWELPWLMQPPL[++++] TWALPWLLEKPF[++++] SPILWLNAPPWA[++++] WHDLWSSNWDTV[++++] GENLMSVGLLRT[++] 6 Phage display (subtractive panning) 3 PMID:37046671 M2 macrophages Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Flow Cytometry Binding of the phage clones to M2 macrophages was analyzed by flow cytometry. +, binding < 25%; ++, binding < 50%; ++++, binding > 80–100%. The 12-mer peptide phage library (Ph.D.™-12) was purchased from New England BioLabs (Ipswich, MA, USA). The phage library was amplified and titered according to the manufacturer’s instructions. The M1 and M2 macrophages used for biopanning were grown in T25 flasks until they reached 80–90% confluency on the day of the experiment. Prior to biopanning, the cells were washed in PBS buffer supplemented with 5% bovine serum albumin (BSA) (washing buffer) to remove dead cells. Thereafter, the phage library (1011 transduction units, TU), diluted in 3 mL washing buffer, was added to the cells and incubated for 1 h at room temperature with gentle agitation. Subsequently, the supernatant was removed and 10 mL washing buffer was added to the cells. The cells were then harvested by gentle scraping and transferred to a 15 mL Falcon tube. The cells were pelleted by centrifugation at 300× g for 3 min and then washed 10 times with the washing buffer to remove unbound phages. Cell-binding phages were eluted in 200 μL elution buffer (0.1 M glycine-HCl, pH2.2, 0.1% BSA) for 10 min at RT with constant rotation followed by centrifugation for 5 min at 12,000× g. The supernatant containing the eluted phages was collected and neutralized with 28 μL neutralization buffer (Tris-HCl, pH 9.2). The eluted phages were amplified in the E. coli ER2738 strain, titered and then used in subsequent rounds of biopanning. In addition, the biopanning was performed with subtraction steps, in which the phage library was pre-incubated with peripheral blood mononuclear cells and/or M1 macrophages prior to affinity selection on M2 macrophages. This pre-incubation step was carried out from round 1. To explore the therapeutic potential of the selected peptides, the M13 phage-displayed peptides were conjugated to the photosensitizer IR700, which has been used for cancer photoimmunotherapy. The phage displaying a dominant peptide (SPILWLNAPPWA) killed both M1 and M2 macrophages, while those displaying the M2-specific peptides killed M2 macrophages only upon near-infrared light exposure. A significant fraction of the M2 macrophages were also killed with the untargeted M13 phage-IR700 conjugates. Hence, M2 macrophages can also be selectively targeted by the wild type M13 phage, which displayed a significant tropism to these cells. The benefits of this photoimmunotherapy include an automatic self-targeting ability of the wild type M13 phage, and the option of genetic manipulation of the phage genome to include tumor targeting peptides, allowing the killing of both M2 macrophages and cancer cells. 3592 LTPHKHHKHLHA(11)[0.4363±0.0133] STPHKHHKHLHA(5)[0.378±0.012] FFADVTKHSQKT(2)[0.2969±0.0243] HPASGWHMRHHR(1)[0.2005±0.0083] 4 Phage display (competitive panning) 4 PMID:35700851 Immunomodulatory protein Ling Zhi-8, LZ-8 mesenchymal stem cells (MSCs) Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorption was measured at 450 nm. Data shown were reproduced from Figure 1A. A specific elution was performed for the fourth round of biopanning, in which the rLZ-8-bound phages were eluted with rLZ-8 polyclonal antibody after washing with TBS-T 0.5% (v/v). rLZ-8 is short for recombinant Ganoderma lucidum immunomodulatory protein-8. The binding mode between of recombinant Ganoderma lucidum immunomodulatory protein-8 (rLZ-8) and the peptide ligand (LTPHKHHKHLHA) was simulated and revealed by molecular docking. Standard addition and repetitive testing were carried out to evaluate the accuracy, reproducibility and feasibility of the developed ELISA detection method. The method based on this peptide ligand was then successfully applied in the quantitative determination of rLZ-8 concentrations in fermentation broth. In summary, the peptide–antigen–antibody sandwich ELISA method developed here could be conveniently applied in the detection of rLZ-8 during fermentation and might provide new insights for the detection of other specific proteins. 3593 TSATKFMMNLSP(24)[0.5012±0.1015] 1 Phage display (subtractive panning) 6 PMID:35323423 Klebsiella pneumoniae KCTC 2208 cell Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The microtiter plates were coated with bacterial cells (1 × 107 colony-forming unit (CFU)/well) overnight at 4 °C and incubated again with 3% BSA in TBS for 1 h at room temperature to block non-specific binding. After washing three times with TBST, one hundred microliters of amplified phages (1 × 109 plaque-forming unit) were added and incubated for 1 h. The wells were washed three times with TBST; subsequently, horseradish peroxidase (HRP)-conjugated anti-M13 monoclonal antibody (1:5000, GE Healthcare, Chicago, IL, USA) was added and incubated for 1 h. The wells were rewashed with TBST, and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) peroxidase substrate (Sigma-Aldrich, St. Louis, MO, USA) was added to the wells to detect phage binding. The color development at 415 nm was recorded using a microplate reader (Synergy, BioTeK, Santa Clara, CA, USA). Data shown were reproduced from Figure 1. To ensure specificity, the phage library was depleted of clones binding to bovine serum albumin (BSA)-coated wells before selecting Klebsiella pneumoniae binding clones. The biopanning-derived peptide TSATKFMMNLSP, KP peptide, displayed a high selectivity for the K. pneumoniae with low cross-reactivity to related Gram-negative bacteria. The specific interaction between KP peptide and K. pneumoniae lipopolysaccharide resulted in the peptide’s selectivity against K. pneumoniae. Quantitative analysis of this interaction by enzyme-linked immunosorbent assay revealed that the KP peptide possessed higher specificity and sensitivity toward K. pneumoniae than commercially available anti-Klebsiella spp. antibodies and could detect K. pneumoniae at a detection limit of e4 CFU/mL. These results suggest that KP peptide can be a promising alternative to antibodies in developing a biosensor system for K. pneumoniae detection. 3594 KWHWKDKNALRM(5/15) GPVNKSSTILRM(3/15) GSLRPGTTNALV(2/15) GLHTSATNLYLH(2/15) AHGNAALVARLK(1/15) FGDLTRGQQRGP(1/15) QGTVARLPIFWP(1/15) 7 Phage display (common panning) 4 PMID:36979401 Pseudomonas aeruginosa ATCC 27853 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The color development in the wells was recorded by FLUOstar Omega microplate reader (BMG LabTech) at the wavelength of 405 nm. Data were not shown. Peptides that bind to P. aeruginosa were selected via biopanning of a 12-mer random phage-displayed peptide library against the bacterial whole cells according to the protocol as described by the manufacturer (New England Biolabs, Ipswich, MA, USA) [28]. For the first round of biopanning, P. aeruginosa ATCC 27853 resuspended in 2 mL of PBS (pH 7.4) was added with 10 µL of the phage library diluted in 100 µL of TBST, followed by incubation at room temperature for 60 min with gentle agitation. Unbound phages were removed by a series of washing with TBST followed by centrifugation. Bound phages were eluted with 200 µL of elution buffer [0.2 M Glycine-HCl (pH 2.2), 1 mg/mL BSA] for 10 min at room temperature, followed by neutralization of the eluted phages with 30 µL of 1 M Tris-HCl (pH 9.1). A titer of the eluted phages was determined by plating an aliquot of the phages on LB IPTG/Xgal agar, while the rest of the phages were amplified in Escherichia coli ER2738. Amplified phages were purified with polyethylene-glycol precipitation, followed by resuspension of the phage pellet in TBS. The titer of the amplified phages was determined before proceeding to the subsequent round of biopanning. A total of four rounds of biopanning were carried out to enrich the clones of phage-displayed peptides binding to P. aeruginosa, with the titers of input phages (phages in TBS after amplification) and output phages (phages in elution buffer) being determined each round. After the final round of biopanning, individual eluted phages on the titer plate were randomly selected for amplification in E. coli ER2738 to be used in phage-ELISA and phage genomic DNA extraction. The affinity-selected peptide (KWHWKDKNALRM) with the highest selection frequency was modified to PAM-5 (KWKWRPLKRKLVLRM) with enhanced antibacterial features by using an online peptide database. Using in vitro microbroth dilution assay, PAM-5 was shown to be active against a panel of Gramnegative bacteria and selected Gram-positive bacteria. Interestingly, the peptide was stable in human plasma by exhibiting a similar bactericidal effect via ex vivo assay. Scanning electron microscopy and SYTOX Green uptake assay revealed that PAM-5 was able to cause membrane disruption and permeabilization of the bacteria. Additionally, the peptide was also able to bind to bacterial DNA as demonstrated by gel retardation assay. In the time-kill assay, PAM-5 was shown to kill the bacteria rapidly in 10 min. More importantly, PAM-5 was non-cytotoxic to Vero cells and non-haemolytic to human erythrocytes at all concentrations tested for the antibacterial assays. 3595 ACEGLYAHWC[0.2756±0.0275] 1 Phage display (common panning) 3 PMID:34259504 Anti-mycophenolic acid (MPA) antibody fragment (Fab), anti-MPA Fab Mycophenolic acid, MPA Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance at 405 nm was measured in a Varioskan plate reader (Thermo Scientific). Data were reproduced from Figure 1A. A commercial phage-displayed peptide library was used to select cyclic peptides that bind to the anti-MPA. The selection rounds were carried out with an automatic magnetic bead processor (KingFisher Thermo Fisher Scientific). See the Supporting Information for antibody coupling to magnetic beads. Briefly, the phage-displayed peptide library (∼2.0 × 1011 phages) was incubated for 2 h with the anti-MPA conjugated beads (50 μg) in a total volume of 505 μL of PBST [PBS, pH 7.4 with 0.05% (v/v) Tween-20]. The beads were subsequently washed twice with PBST for 30 s, and then the bound phages were eluted with 100 μL of 0.1 M triethylamine (pH 11.2) for 30 min. The resulting solution containing the eluted phages was immediately neutralized with 70 μL of 1 mol L–1 Tris-HCl (pH 6.8). Amplification of the eluted phages was carried out by adding 70 μL of the eluate to a 40 mL early-log phase ER2738 culture in LB and incubating at +37 °C for 4.5 h. The cells were harvested by centrifugation (10 min, 12,000g, +4 °C), and the supernatant was collected. The amplified phages were precipitated overnight at +4 °C after adding to the supernatant 1/6 volume of 20% poly(ethylene glycol) (PEG)/2.5 mol L–1 NaCl. Then, the precipitated phages were collected by centrifugation (15 min, 12,000g, +4 °C) and resuspended in 3 mL of PBS. The precipitation was repeated with 20% PEG/2.5 mol L–1 NaCl on ice for 1 h, followed by centrifugation (10 min, 12,000g, +4 °C). Finally, the pellet containing the phages was resuspended in 500 μL of PBS. The amplified phage solution was utilized for the consequent selection round. After identifying the best mycophenolic acid (MPA) mimetic (ACEGLYAHWC with a disulfide constrained loop), several immunoassay approaches were tested, and a recombinant fusion protein containing the peptide sequence with a bioluminescent enzyme, NanoLuc, was developed. The recombinant fusion enabled its direct use as the tracer in competitive immunoassays without the need for secondary antibodies or further labeling. A bioluminescent sensor, using streptavidin-coupled magnetic beads for the immobilization of the biotinylated Fab antibody, enabled the detection of MPA with a detection limit of 0.26 ng mL−1 and an IC50 of 2.9 ± 0.5 ng mL−1. The biosensor showed good selectivity toward MPA and was applied to the analysis of the immunosuppressive drug in clinical samples, of both healthy and MPA-treated patients, followed by validation by liquid chromatography coupled to diode array detection. 3596 AAHRVGGFNYHM(7/20)[1097.7413±82.1182] SVPLNSWSIFPR(5/20)[1526.2958±108.2934] STVYHTTPYHNR(3/20)[NT] 3 Phage display (common panning) PMID:37098917 Staphylococcus aureus, S.aureus Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA An ELISA plate was coated with an S. aureus suspension using the methods described in “Two panning methods for phage selection,” above. Serial dilutions of the phage in 100 μL of TBST were prepared, added to each well, and then incubated at RT for 1 h with agitation. The plate was washed six times with TBST; then, 200 μL of the diluted HRP-conjugated anti-M13 monoclonal antibody (GE Healthcare) was added to each well and incubated at RT for 1 h with agitation. The HRP substrate solution was prepared according to the manufacturer’s instructions. Finally, 200 μL of substrate solution was added to each well and incubated for 60 min at room temperature (RT) with gentle agitation, and the plate was read at 405 to 415 nm. Staphylococcus aureus cells were cultured in Luria-Bertani (LB) medium until the optical density at 600 nm (OD600) reached 0.6 to 0.8. The cells were then collected by centrifugation, washed with phosphate-buffered saline (PBS), and mixed with ethanol before being dried on a 96-well plate (100 μL/well) at room temperature (RT) overnight. For the first panning round, the cells fixed on the 96-well plate were exposed to e11 PFU/mL of phage library in TBST buffer (Tris-buffered saline with 0.05% Tween 20, pH 7.5) (100 μL/well). Unbound phages were eliminated by washing five times with TBST, and the bound phages were eluted with 100 μL elution buffer (0.2 M Glycine-HCl) for 20 min at room temperature. The eluted phage was neutralized with 1 M Tris-HCl (pH 9.1) and amplified in E. coli ER2738, followed by purification with polyethylene glycol precipitation. A phage clone displaying a peptide capable of specific binding to a whole S. aureus cell was selected from a 12-mer phage peptide library. The peptide sequence was SVPLNSWSIFPR. The selected phage’s ability to bind specifically with S. aureus was confirmed using an enzyme-linked immunosorbent assay, and the chosen peptide was then synthesized. The results showed that the synthesized peptides displayed high affinity with S. aureus but low binding ability with other strains, including Gram-negative and Gram-positive bacteria such as Salmonella sp., Shigella spp., Escherichia coli, and Corynebacterium glutamicum. In addition, yeast vacuoles were used as a drug carrier by encapsulating daptomycin, a lipopeptide antibiotic used to treat Gram-positive bacterial infections. The expression of specific peptides at the encapsulated vacuole membrane created an efficient system that can specifically recognize and kill S. aureus bacteria.  3597 VGQFGTSQMILP(6/20)[1168.5683±108.2934] SWPTFTVLKNHA(3/20)[NT] NFTLQAHPHKYP(3/20)[NT] 3 Phage display (common panning) PMID:37098917 Staphylococcus aureus, S.aureus Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA An ELISA plate was coated with an S. aureus suspension using the methods described in “Two panning methods for phage selection,” above. Serial dilutions of the phage in 100 μL of TBST were prepared, added to each well, and then incubated at RT for 1 h with agitation. The plate was washed six times with TBST; then, 200 μL of the diluted HRP-conjugated anti-M13 monoclonal antibody (GE Healthcare) was added to each well and incubated at RT for 1 h with agitation. The HRP substrate solution was prepared according to the manufacturer’s instructions. Finally, 200 μL of substrate solution was added to each well and incubated for 60 min at room temperature (RT) with gentle agitation, and the plate was read at 405 to 415 nm. The e11 PFU/mL of Ph.D.-12 phages was exposed to S. aureus cells (OD600, 0.5) in PBS for 60 min at RT with gentle agitation. The bacteria-phage mixture was centrifuged for 5 min at 16,000 × g, and then the unbound phages were removed by washing 10 times (centrifugation at 16,000 × g for 5 min with 1 mL TBST). The pellet that contained the bound phages was eluted with 250 μL of 0.2 M glycine-HCl (pH 2.2) with slight shaking at RT for 20 min and was then sonicated for 10 min with 50% amplitude, 30 s on and 30 s off. Subsequently, the solutions were neutralized with 25 μL of 1 M Tris-HCl (pH 9). The eluate (10 μL) would be used for phage titering, and 500 μL was used to infect E. coli ER2738 for amplification. A phage clone displaying a peptide capable of specific binding to a whole S. aureus cell was selected from a 12-mer phage peptide library. The peptide sequence was SVPLNSWSIFPR. The selected phage’s ability to bind specifically with S. aureus was confirmed using an enzyme-linked immunosorbent assay, and the chosen peptide was then synthesized. The results showed that the synthesized peptides displayed high affinity with S. aureus but low binding ability with other strains, including Gram-negative and Gram-positive bacteria such as Salmonella sp., Shigella spp., Escherichia coli, and Corynebacterium glutamicum. In addition, yeast vacuoles were used as a drug carrier by encapsulating daptomycin, a lipopeptide antibiotic used to treat Gram-positive bacterial infections. The expression of specific peptides at the encapsulated vacuole membrane created an efficient system that can specifically recognize and kill S. aureus bacteria.  3598 LPSYNLHPHVPP(3/12) IPLLNPGSMQLS(4/12) 2 Phage display (subtractive panning) 2 PMID:36303134 Human lens capsules from patients with exfoliation syndrome (XFS) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Human lens capsules collected from patients without exfoliation syndrome (XFS) were used for subtractive screening. peptides differentiate between exfoliative and non-affected regions of the human lens capsule 3599 LPSYNLHPHVPP(11/52) IPLLNPGSMQLS(37/52) 2 Phage display (subtractive panning) 3 PMID:36303134 Human lens capsules from patients with exfoliation syndrome (XFS) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Human lens capsules collected from patients without exfoliation syndrome (XFS) were used for subtractive screening. peptides differentiate between exfoliative and non-affected regions of the human lens capsule 3600 HDYLYYTFTGNP(4)[1.1555±0.0301] TKFSPPSFWYLH(2)[1.3806±0.0245] GPFWPTQGAHLR(2)[NT] WTRKYMPYGPTP(2)[NT] GPGLMLRPAFSN(1)[NT] ETDYHWMNYLFS(1)[NT] HAHVIYSPHLPP(1)[NT] AVTLTQLSPQIH(1)[NT] 8 Phage display (common panning) 4 PMID:33340637 Aminopeptidase Ey Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured at 490 nm after using OPD color reagent for 10 min. Biopanning of the phage was performed using the Ph.D™-12 Phage Display Peptide Library Kit (New England Biolabs, Beverly, MA, USA) according to the manufacturer's instructions with minor modifications. In brief, ELISA plates were coated with rgAPN protein (2 μg/ml) per well overnight at 4 °C. The original phage library (1.5 × 1011 pfu/well) was incubated. Phages in 1st round biopanning were obtained by eluting (0.2 mol/L Glycine-HCl), and then neutralized (1 mol/L Tris-HCl). The treatments were replicated 4 times. Titers of phages for each round were determined on LB/IPTG/Xgal (Low LB + 0.1% [v/v] IPTG/Xgal) plates. High-affinity ligands of pET-gAPN protein, H (HDYLYYTFTGNP) and T (TKFSPPSFWYLH), screened by the phage display peptide library, reduced IBV infection in vivo and in vitro via blocking the key antigenic motifs, and might provide a novel countermeasure for IB prevention or treatment. In addition, the results indirectly demonstrated that gAPN acts as a receptor for IBV. 3601 TWWNPRLVYFDY(32)[248.506±48.1295] GVWWTWGNYGQM(5)[99.1517±25.2107] YTPNWHFRWMPA(2)[82.7265±48.3205] APTTWFNSDSIT(2)[13.5881±14.3243] 4 Phage display (common panning) 3 PMID:34962367 Graphene paper, GP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) To assess the binding affinity of GP-binding peptides, four selected GP-binding phages (with wild-type M13 phage as a control group) were amplified by infecting E. coli. After GP containers were UV treated and washed by TBS buffer, 200 μL of the corresponding phage solution (1010 pfu/ mL) was added into five containers separately and incubated for 60 min. The unbound phages were washed away with TBST. To get the GP-bound phages out of the GP substrates, 240 μL of the elution buffer was allowed to interact with the substrates for 8 min. Finally, 40 μL of neutralizer was added, and 10 μL of resultant was titrated to quantify the phages. First, GP was folded into a container and exposed to ultraviolet irradiation for an hour. Then, the container was washed using TBS. The phage library (2.0e11 pfu) was added to the GP container for incubation. After 1 h of incubation, the unbound phages were washed away with TBST. To elute bound phages, 240 μL of glycine-HCl (0.2 M, pH 2.2) was added and incubated for 8 min. Then the eluate was neutralized with 40 μL of Tris-HCl (1 M, pH 9.1). Then, 10 μL of the eluate was titrated. The rest of the eluate was amplified in ER2738 E. coli culture and treated as a sublibrary for the next round of biopanning. Output phages in round 3 were sequenced to identify the sequences of foreign peptides displayed at their protein 3 (p3) tips. We first screened a high-affinity graphene paper (GP) binding peptide (TWWNPRLVYFDY) by the phage display technique. Then we chemically conjugated the GP-binding peptide to the synthetic hydroxyapatite (HA) nanorods. The GP-binding peptide on the resultant HA nanorods enabled them to be bound and assembled onto the GP substrate with high affinity, forming a GP-peptide-HA composite with significantly improved hydrophilicity of GP. The composite promoted the attachment and proliferation of mesenchymal stem cells (MSCs), demonstrating its outstanding biocompatibility. Due to the unique compositions of the composite, it was also found to induce osteogenic differentiation of MSCs in vitro in the absence of other inducers in the medium, by verifying the expression of the osteogenic markers including collagen-1, bone morphogenetic proteins 2, runx-related transcription factor 2, osteocalcin, and alkaline phosphatase. Our work suggests that the GP-binding peptide can be used to link inorganic nanoparticles onto GP to facilitate the biomedical applications of GP. 3602 ACGTKPTKFC(6/26)[1.3898±0.0246] ACPTTSTQYC(5/26)[0.8509±0.0099] ACTDKASSSC(5/26)[0.7775±0.0246] ACTPRSANYC(3/26)[1.0217±0.0915] ACLKTYWYNC(2/26)[1.2293±0.0772] 5 Phage display (common panning) 4 PMID:37230729 Nucleoprotein, N Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA SARS-CoV-2 NP (1 μg/mL) was fixed in the wells of the microplates overnight. peptides are powerful biomolecular tools for SARS-CoV-2 detection, providing a new and inexpensive method of rapidly screening infections as well as rapidly diagnosing coronavirus disease 2019 patients 3603 SFYDFEMQGFFI(4)[2.6799±0.2464] SFYDEMGFI(4)[3.0158±0.2156] SYFMQFF(3)[3.1003±0.1193] 3 Phage display (common panning) 3 PMID:34170090 Anti-infectious bronchitis virus (IBV) M41 strain S1 protein monoclonal antibody 3D9 Spike protein S1 of Spike glycoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance at 450 nm was measured using a Bio-Rad Microplate Reader. A standard biopanning procedure was carried out according to the manufacturer's instructions. Briefly, one well of a 96-well microtiter plate was coated with 100 µL of mAb 3D9 with a final concentration of 100 µg/mL. After blocking, 100 µL of the phages (about 2.0e11 pfu/mL) from the Ph.D.-12 phage library was added to the wells and incubated for 1 h at room temperature. The unbound phages were removed, and the wells were washed with 0.1%TBST. An elution buffer was added to each well with gentle shaking for 1 h at room temperature. The eluent containing the phages was mixed with a neutralization buffer to determine the titer of the phages and for amplification. The coated mAb 3D9 concentration (100 µg/mL, 75 µg/mL, and 50 µg/mL) was reduced and the concentration of Tween-20 in TBST buffer (0.1%, 0.2%, and 0.3%) was increased according to the number of biopanning steps. Sequence analysis showed that the dominant sequence was SFYDFEMQGFFI. Indirect competitive enzyme-linked immunosorbent assay showed that SFYDFEMQGFFI is a mimotope of the S1 protein that was predicted by PepSurf. The mimotope may provide information for further structural and functional analyses of the S1 protein. 3604 NFWISPKLAFAL(6/24)[1.2595±0.0186] AEAWTGFSASGV(2/24)[0.1887±0.0186] NHHYTYHQYTVG(2/24)[0.4545±0.0165] HVNYGMHGYETT(4/24)[0.359±0.0304] NTACDMSGRHCH(3/24)[0.4685±0.0469] SSFGMSSQLISH(1/24)[0.1211±0.0068] WDMWPSMDWKAE(3/24)[0.1748±0.0068] QINPEFTPTMRW(1/24)[0.2306±0.0072] 8 4 PMID:35948070 Spike protein S1 of spike glycoprotein Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The optical density of each well at 452 nm (OD452nm) was measured. The bovine serum albumin (BSA) negative selection was added to rounds 2, 3, and 4 biopanning. That is, the phage was incubated with BSA for 4 h before incubating the phage with SARS-CoV-2 S1. After four rounds of biopanning from the pIII phage display library, a phage monoclone expressing the NFWISPKLAFAL peptide was identified, which had the best affinity and selectivity for SARS-CoV-2 S1 with a dissociation constant of 3.45 ± 0.58 nM.  3605 DGFIRPSGVRVA(19/78)[0.6748±0.4406] WDMWPSMDWKAE(8/78)[1.5887±0.1874] DGSMLNRMRGFS(3/78)[0.3231±0.0985] 3 Phage display (common panning) 4 PMID:36944950 Cathepsin B Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured at 405 nm using a Multiskan FC microplate photometer (Thermo Scientific, Waltham, MA, USA). Data shown were reproduced from Figure 2A. Biotinylated cathepsin B (99 μL, 500 nM) was pre-reacted with the Ph.D.-12 random phage library (1 μL, 1.0 × 1013 PFU/mL) at 100 rpm for 1 h. Subsequently, 100 µL of the complex mixture was added onto a pre-washed streptavidin-coated microplate, and allowed to react at a shaking speed of 100 rpm for 10 min to facilitate specific binding between avidin and biotin. Then, 0.1 mM biotin was added as a blocking agent and allowed to react for 5 min. After removing the unbound phages and residual biotin, the plate was washed 10 times with 0.1% PBST (0.1 M phosphate-buffered saline (PBS) with 0.1% Tween 20), and the bound phages were eluted using 100 µL of 0.2 M glycine–HCl (pH 2.2) with 1 mg/mL of bovine serum albumin (BSA) solution. Finally, the eluent was neutralized with 15 µL of Tris–HCl (pH 9.1) to prevent deactivation or destruction of the desired phages. Throughout this process, the desired phages were amplified using Escherichia coli ER2738 making sufficient copies for the next rounds. The amplified phages were then harvested by polyethylene glycol (PEG)/NaCl precipitation (20% (v/v) PEG-8000 with 2.5 M NaCl). After every biopanning round, the phages were tittered, and the displayed peptide sequences were analyzed using the − 96 gIII sequencing primer (5ʹ-HOCCC TCA TAG TTA GCG TAA CG-3ʹ). A phage-display library was biopanned against biotinylated cathepsin B to identify a high-affinity peptide with the sequence WDMWPSMDWKAE. The identified peptide-displaying phage clones and phage-free synthetic peptides were characterized using enzyme-linked immunosorbent assays (ELISAs) and electrochemical analyses (impedance spectroscopy, cyclic voltammetry, and square wave voltammetry). Feasibilities of phage-on-a-sensor, peptide-on-a-sensor, and peptide-on-a-AuNPs/MXene sensor were evaluated. 3606 AHNHTPIKQKYL(2/55)[512.103±48.2444] ANTELALANRKH(2/55)[352.5796±17.2553] NWGVMPWIGATT(2/55)[389.5553±13.3817] 3 Phage display (common panning) 4 PMID:32971483 Tyrosine-protein kinase receptor UFO Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) EIS was carried out with a DC potential of 0.2 V using an alternating voltage of 10 mV in a frequency range of 10 Hz to 10 kHz. The AXL protein was dissolved in PBS buffer and subsequently transferred to a microwell plate. Following overnight incubation at 4 °C, the wells were washed with PBS buffer to remove the unbound AXL protein and then incubated with blocking buffer at 4 °C for 1 h. After removing the residual blocking solution, the wells were washed six times with PBST buffer (0.1 M PBS buffer with 0.1% Tween 20). The Ph.D.-12 peptide library (1 × 1011 plaque forming units [PFUs]) was added to the wells containing immobilized AXL protein and binding was performed by incubating the plate at 25 °C with shaking. Following successful shaking, the plate was washed 10 times with PBST to remove the unbound phage particles or residual protein and the AXL-bound phage particles were finally eluted using an elution buffer (0.2 M glycine-HCl, pH 2.2). To minimize non-specific binding during the entire biopanning process, the Tween 20 concentration used in the washing step of round 1, 2, and 3–5 was increased to 0.1, 0.3, and 0.5%, respectively. The elution fraction containing the most AXL-bound phage particles was immediately neutralized using Tris-HCl (pH 9.1) to prevent their activity. The neutralized eluted phage fraction was used to infect E. coli ER2738 to amplify sufficient copies of the peptides beyond preparation of DNA sequencing and next round of panning. The phage titer was determined using Luria-Bertani broth containing isopropyl β-D-thiogalactopyranoside and X-gal, and PFU was calculated by counting single blue plaques after overnight incubation at 37 °C. Biopanning of M13 phage library successfully identified a high affinity peptide, with the sequence AHNHTPIKQKYL. To study the feasibility of using free peptides for molecular recognition, we synthesized a series of amino acid-substituted peptides and examined their binding affinity for a tyrosine kinase receptor (AXL) using electrochemical impedance spectroscopy and square wave voltammetry. Most synthetic peptides had non-identical random coil structures based on circular dichroism spectroscopy. Of the peptides tested, AXL BP1 (AHNHTPIKQKYL) exhibited nanomolar binding affinity for AXL. To verify whether AXL BP1 could be used as a peptide inhibitor at the cellular level, two functional tests were carried out: a WST assay for cell viability and qRT-PCR for quantification of RNA levels in Zika virus-infected Huh7 cells. The results showed that AXL BP1 had low cytotoxicity and could block Zika virus entry. These results indicate that newly identified affinity peptides could potentially be used for the development of Zika virus entry inhibitors. 3607 WSLGYTG(4/20) GTIYWNS(1/20) SPLSPRY(1/20) ETGITRQ(1/20) WIFTPLG(1/20) RNSWPVW(1/20) GSSGKPG(1/20) RGTGHYW(1/20) WWSTHDR(1/20) RNMRGYG(1/20) WTARPTG(1/20) GSWTTGQ(1/20) 12 3 PMID:33481966 T-cell surface glycoprotein CD3 zeta chain Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Flow Cytometry All candidate phage clones harvested from the biopanning experiments and two random clones from the stock library were amplified, tittered, and plated for the determination of each concentration (PFU/ml). Streptavidin-coated iron-oxide microbeads (Dynabeads M280, Invitrogen) were decorated with biotinylated human CD3e (Avitag) at a saturation density (>10 μg of biotinylated protein per 1 mg of Dynabeads). CD3e-coated beads were washed with PBS buffer once and incubated with candidate phage clones (10^12 virions per 1 mg beads) in 200 μl TBST buffer (tris-buffered saline with 0.1% tween) for 30 minutes at room temperature on a shaker. After washing the beads with FACS buffer once, the beads were incubated with anti-M13 Major Coat Protein antibody (1 : 100 dilution) for 30 minutes at room temperature on a shaker. The bead samples were finally washed and resuspended in FACS buffer to be examined using flow cytometry. Screening of the PhD-7 phage display peptide library on human recombinant CD3ε protein as the target protein was carried out according to the manufacturer's protocol with some modifications. Briefly, 50 μl of a 50% aqueous suspension of Pierce protein A/G magnetic beads was washed with 1 ml of TBST washing buffer (tris-buffered saline with 0.1% tween) on a magnetic separator and blocked in blocking buffer (0.1 M NaHCO3 (pH 8.6) with 0.5% (w/v) BSA) for 1 h at 4 °C on a rotator. 2 picomoles of target protein was mixed with 1011 pfu (plaque-forming unit) of the phage library in 200 μl of TBST buffer and incubated for 15 minutes at room temperature on a rotator and then incubated with pre-washed and pre-blocked magnetic beads for 15 minutes at room temperature on a rotator. After the microbeads were washed vigorously 15 times with 1 ml TBST, the bound phage was eluted by 1 ml of glycine buffer (0.2 M glycine–HCl (pH 2.2) + 0.1% (w/v) BSA) and neutralized by the addition of 150 μl of 1 M Tris–HCl (pH 9.1). The harvested phages were amplified by infecting E. coli (ER2738) cells and precipitated using PEG8000/NaCl solution. After each round, a negative selection was performed by incubating the harvested phage clones from the previous round with the empty protein A/G magnetic beads in TBST buffer for 20 minutes at room temperature on a rotator. Only the unbound phages in this negative selection were used for the next rounds of panning. After the third and the fifth round of biopanning, the resulting phage clones tittered on LB/IPTG/Xgal plates, and 20 out of 100 blue plaques were randomly selected for sequencing. WSLGYTG demonstrated a superior binding behavior to other clones in the binding assays against recombinant T-cell surface glycoprotein CD3 zeta chain (CD3 zeta) on microbeads or Jurkat cells. The synthesized peptide also showed specific binding to Jurkat cells in a dose-dependent manner but not to B cell lymphoma line, 2PK3 cells. Molecular modeling and docking simulation confirmed that the selected peptide ligand in an energetically stable conformation binds to a pocket of CD3 zeta that is not hidden by either CD3 gamma (T-cell surface glycoprotein CD3 gamma chain) or CD3 delta (T-cell surface glycoprotein CD3 delta chain). Lastly, magnetic microbeads conjugated with the synthesized peptide ligands showed a weak but specific association with Jurkat cells and induced the calcium flux, a hallmark indication of proximal T cell receptor signaling, which gave rise to an enhancement of IL-2 section and cell proliferation. The novel peptide ligand and its various multivalent forms have a great potential in applications related to T cell biology and T cell immunotherapy. 3608 WSLGYTG(18/20) GSWTTGQ(1/20) YNHTMMY(1/20) 3 5 PMID:33481966 T-cell surface glycoprotein CD3 zeta chain Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Flow Cytometry All candidate phage clones harvested from the biopanning experiments and two random clones from the stock library were amplified, tittered, and plated for the determination of each concentration (PFU/ml). Streptavidin-coated iron-oxide microbeads (Dynabeads M280, Invitrogen) were decorated with biotinylated human CD3e (Avitag) at a saturation density (>10 μg of biotinylated protein per 1 mg of Dynabeads). CD3e-coated beads were washed with PBS buffer once and incubated with candidate phage clones (10^12 virions per 1 mg beads) in 200 μl TBST buffer (tris-buffered saline with 0.1% tween) for 30 minutes at room temperature on a shaker. After washing the beads with FACS buffer once, the beads were incubated with anti-M13 Major Coat Protein antibody (1 : 100 dilution) for 30 minutes at room temperature on a shaker. The bead samples were finally washed and resuspended in FACS buffer to be examined using flow cytometry. Screening of the PhD-7 phage display peptide library on human recombinant CD3ε protein as the target protein was carried out according to the manufacturer's protocol with some modifications. Briefly, 50 μl of a 50% aqueous suspension of Pierce protein A/G magnetic beads was washed with 1 ml of TBST washing buffer (tris-buffered saline with 0.1% tween) on a magnetic separator and blocked in blocking buffer (0.1 M NaHCO3 (pH 8.6) with 0.5% (w/v) BSA) for 1 h at 4 °C on a rotator. 2 picomoles of target protein was mixed with 1011 pfu (plaque-forming unit) of the phage library in 200 μl of TBST buffer and incubated for 15 minutes at room temperature on a rotator and then incubated with pre-washed and pre-blocked magnetic beads for 15 minutes at room temperature on a rotator. After the microbeads were washed vigorously 15 times with 1 ml TBST, the bound phage was eluted by 1 ml of glycine buffer (0.2 M glycine–HCl (pH 2.2) + 0.1% (w/v) BSA) and neutralized by the addition of 150 μl of 1 M Tris–HCl (pH 9.1). The harvested phages were amplified by infecting E. coli (ER2738) cells and precipitated using PEG8000/NaCl solution. After each round, a negative selection was performed by incubating the harvested phage clones from the previous round with the empty protein A/G magnetic beads in TBST buffer for 20 minutes at room temperature on a rotator. Only the unbound phages in this negative selection were used for the next rounds of panning. After the third and the fifth round of biopanning, the resulting phage clones tittered on LB/IPTG/Xgal plates, and 20 out of 100 blue plaques were randomly selected for sequencing. WSLGYTG demonstrated a superior binding behavior to other clones in the binding assays against recombinant T-cell surface glycoprotein CD3 zeta chain (CD3 zeta) on microbeads or Jurkat cells. The synthesized peptide also showed specific binding to Jurkat cells in a dose-dependent manner but not to B cell lymphoma line, 2PK3 cells. Molecular modeling and docking simulation confirmed that the selected peptide ligand in an energetically stable conformation binds to a pocket of CD3 zeta that is not hidden by either CD3 gamma (T-cell surface glycoprotein CD3 gamma chain) or CD3 delta (T-cell surface glycoprotein CD3 delta chain). Lastly, magnetic microbeads conjugated with the synthesized peptide ligands showed a weak but specific association with Jurkat cells and induced the calcium flux, a hallmark indication of proximal T cell receptor signaling, which gave rise to an enhancement of IL-2 section and cell proliferation. The novel peptide ligand and its various multivalent forms have a great potential in applications related to T cell biology and T cell immunotherapy. 3609 LTPHKHHKHLHA(35)[+] RLATYPQLSVTQ(NA)[+] 2 Phage display (common panning) 3 PMID:34111485 Anti-bovine viral diarrhea virus (BVDV) serum Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Western blot BSA-conjugated peptides were separated by SDS-PAGE, transferred onto polyvinylidene fluoride (PVDF) membranes (Bio-Rad, Hercules, CA, USA). After blocking with 5% non-fat milk in PBST for 1 h at room temperature, the membranes were incubated with goat anti-BVDV specific antibody overnight at 4 ◦C, and donkey anti-goat IgG-HRP for 1 h at room temperature. After extensive washing with PBST, the reactive protein bands were developed using Clarity Western ECL substrate (BioRad, cat# 1705061). The phage library panning was performed following the protocol specified by the vendor with modification. In brief, 60 mm dishes were coated with commercial goat anti-BVDV serum (1:100 dilution) in 0.1 M NaHCO3 (pH 8.6) overnight at 4 °C. The coated dishes were blocked with 5 mg/ml BSA in 0.1 M NaHCO3 (pH 8.6) for 2 h at 4 °C. After rapid washing 6 times with TBST (TBS + 0.1% [v/v] Tween-20), the dishes were incubated with 1 × 1011 clones of the phage from Ph.D.-12 phage display library in 1 ml of TBST for 30 min at room temperature with gentle rocking. Unbound phages were washed away by 10 times of washing with TBST at room temperature. Bound phages were eluted with 1 ml of 0.2 M Glycine-HCl (pH 2.2) containing 1 mg/ml BSA with gentle rocking for 15 min, then neutralized with 150 μl of 1 M Tris-HCl, pH 9.1. Recovered phages were amplified by incubation with 20 ml of E. coli ER2738 (OD600 0.01–0.05) with vigorous shaking for 4.5 h at 37 °C. The amplified phages, were purified by precipitation with 1/6 volume of 20% PEG/2.5 M NaCl overnight at 4 °C, and centrifugation for 30 min at 10,000g. The purified phages were taken over additional binding/amplification/purification cycles to enrich the pool in favor of binding sequences. After 3 rounds of panning, 100 individual clones are characterized by DNA sequencing with primer 28gIII primer (5′-GTA TGG GAT TTT GCT AAA CAA C-3′) as described in the manufacturer's protocol.  We screened a 12-mer phage display peptide library using commercial goat anti-bovine viral diarrhea virus (BVDV) serum, and identified a mimotope “LTPHKHHKHLHA” referred to as P3. With sequence alignment, a putative B-cell epitope “77ESRKKLEKALLA88” termed as P3-BVDV1/2 residing in BVDV core protein was identified. The synthesized peptides of both P3 and P3-BVDV1/2 show strong reactivity with BVDV serum in immune blot assay. Immunization of mice with these individual peptides leads to the production of antibody that cannot neutralize virus infectivity. 3610 CLNTPLKSC(6)[1.038±0.1546] CQPGSLGSC(2)[0.7684±0.0491] CGPRAATSC(1)[0.6455±0.0903] CYKPNQTWC(1)[0.9324±0.0903] CQDPPAAPC(1)[1.0148±0.1886] CFNGIAAEC(1)[0.842±0.1728] CTKPAVKFC(1)[0.6375±0.0729] 7 Phage display (subtractive panning) 3 PMID:35408679 Insulinoma cell line MIN6 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The 96-well plates were then measured at 450 nm using an ELISA reader (Bio-Tek ELX 800, Vermont, NE, USA). After the first round of phage screening on mice insulinoma cell line MIN6, the eluted phage was added to the blocked murine pancreatic cancer cell line KPC cells to absorb the phages, which can combine with KPC cells. The remaining phages (not absorbed by KPC cells) were collected for amplification. Three rounds of phage screening were performed on MIN6 (positive selection) and KPC cells (negative selection). We used phage display libraries to screen a beta-cell-targeted peptide, LNTPLKS, which was tagged with fluorescein isothiocyanate (FITC). This peptide was validated for targeting beta-cell with in vitro and in vivo studies. Immunocytochemistry (ICC) and fluorescence-activated cell sorting (FACS) analysis were used to validate the target specificity of the peptide. FITC-LNTPLKS displayed much higher fluorescence in beta cells vs. control cells in ICC. This discrimination was consistently observed using primary rodent islet. FACS analysis showed right shift of peak point in beta cells compared to control cells. The specific bind to in situ islet was verified by in vitro experiments using rodent and human pancreatic slices. The peptide also showed high affinity of islet grafts under the renal capsule. In the insulinoma animal model, we could find FITC-LNTPLKS accumulated specifically to the tumor, thus indicating a potential clinical application of molecular imaging of insulinoma. 3611 DHAQRYGAGHSG(20/30) 1 Phage display (common panning) 2 PMID:36177466 Recombinant receptor-binding domain (RBD) of glycoprotein protein Angiotensin-converting enzyme 2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Optical absorbance was measured at 450 nm in a microplate reader (Synergy H1, BioTek Instruments, Inc.,Winooski, VT, United States). Data shown were preproduced from Figure 1. In the competitive S-trimer-ACE2-binding experiments, synthetic C2 (DHAQRYGAGHSG) peptide inhibited S-trimer binding onto 293T-ACE2hR cells at high concentrations (50 mM) but not at lower concentrations (10 mM and below), neither for the settings of S-trimer binding onto recombinant ACE2 proteins. 3612 DHAQRYGAGHSG(16/25) 1 Phage display (common panning) 3 PMID:36177466 Recombinant receptor-binding domain (RBD) of glycoprotein protein Angiotensin-converting enzyme 2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Optical absorbance was measured at 450 nm in a microplate reader (Synergy H1, BioTek Instruments, Inc.,Winooski, VT, United States). Data shown were preproduced from Figure 1. In the competitive S-trimer-ACE2-binding experiments, synthetic C2 (DHAQRYGAGHSG) peptide inhibited S-trimer binding onto 293T-ACE2hR cells at high concentrations (50 mM) but not at lower concentrations (10 mM and below), neither for the settings of S-trimer binding onto recombinant ACE2 proteins. 3613 DHAQRYGAGHSG(7/30) HWKAVNWLKPWT(6/30) 2 Phage display (common panning) 2 PMID:36177466 Spike glycoprotein trimer, S-trimer Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Optical absorbance was measured at 450 nm in a microplate reader (Synergy H1, BioTek Instruments, Inc.,Winooski, VT, United States). Data shown were preproduced from Figure 1. In the competitive S-trimer-ACE2-binding experiments, synthetic C2 (DHAQRYGAGHSG) and C6 (HWKAVNWLKPWT) peptides inhibited S-trimer binding onto 293T-ACE2hR cells at high concentrations (50 mM) but not at lower concentrations (10 mM and below), neither for the settings of S-trimer binding onto recombinant ACE2 proteins. 3614 DHAQRYGAGHSG(10/25) HWKAVNWLKPWT(10/25) 2 Phage display (common panning) 3 PMID:36177466 Spike glycoprotein trimer, S-trimer Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Optical absorbance was measured at 450 nm in a microplate reader (Synergy H1, BioTek Instruments, Inc.,Winooski, VT, United States). Data shown were preproduced from Figure 1. In the competitive S-trimer-ACE2-binding experiments, synthetic C2 (DHAQRYGAGHSG) and C6 (HWKAVNWLKPWT) peptides inhibited S-trimer binding onto 293T-ACE2hR cells at high concentrations (50 mM) but not at lower concentrations (10 mM and below), neither for the settings of S-trimer binding onto recombinant ACE2 proteins. 3615 LTRIKW(20%)[0.86] LTRMRY(20%)[0.46] FAPRWR(6.66%)[1.22] FMLRRL(6.66%)[1.13] MYIRPR(6.66%)[0.85] FARVRQ(6.66%)[0.71] LSRMPK(3.33%)[1.15] FSRHPR(3.33%)[1.23] MMRWKS(3.33%)[1.17] MILRKT(3.33%)[0.71] FTRLRA(3.33%)[0.97] LTLRRL(3.33%)[0.75] FGRVPR(3.33%)[0.56] FAFRPK(3.33%)[0.3] MSRMTR(3.33%)[0.56] LTWRTY(3.33%)[0.4] 16 Phage display (common panning) 3 PMID:35306102 Avian influenza virus (H9N2) particles Not determined. Not determined. Not determined. X6 fdMED1 phage display library ELISA Absorbance was measured at 450 nm after subtraction of background and shown. Affinity selection of H9N2-binding peptides was performed by direct biopanning the virus with a phage library displaying linear hexapeptides (Houimel et al., 1999). Purified H9N2 avian influenza virus (15µg/ml in PBS pH 7.4) was coated overnight at 4°C on a NUNC Maxisorbant microplate (NUNC, Danemark), then wells were washed three times with PBS pH 7.4 and saturated 2h at room temperature with 200  μl of PBS-BSA 1%. Phage-peptides (1012 tu/ml in 200  μl PBS-BSA 1%) were added to each well and then incubated for 2h at room temperature. Unbound phages were removed by washing the wells ten times with PBS containing 0.05% Tween-20 (PBST), ten further times with PBS alone, and specifically bound phage was eluted with 100 µl of 100  mM triethylamine, pH 12, immediately buffered with 1 ml of 1 M Tris-HCl, pH 7.4. Eluated phages were propagated in exponentially growing E. coli TG1 and grown overnight at 30°C selectively with 15 µg/ml tetracycline. The produced phage particles were precipitated from the bacterial supernatant with 20% PEG 6000/2.5 M NaCl on ice for 2 h and pelleted by centrifugation for 20 min at 4000 x g. The number of recovered phage was calculated for each of three rounds of panning with decreasing numbers of H9N2 virus after each round. Sixteen different phage-peptides were able to bind specifically the H9N2 virus, among them, 13 phage-peptides interacted with the hemagglutinin H9. Two selected peptides, P1 (LSRMPK) and P2 (FAPRWR) have shown antiviral activity in ovo and P1 was more protective in vivo then P2 when co-administered with the H9N2 virus. Mechanistically, these peptides prevent infection by inhibiting the attachment of the H9N2 virus to the cellular receptor. Molecular docking revealed that the peptides LSRMPK and FAPRWR bind to hemagglutinin protein H9, but interact differently with the receptor binding site (RBS). The present study demonstrated that the peptide P1 (LSRMPK) could be used as a new inhibitory molecule directed against the H9N2 virus. 3616 NQILSLLGI[no binding, no binding] ACHAWAPTR[5.14, 6] AKKIILRLS[no binding, no binding] 3 Phage display (common panning) 3-4 PMID:37191335 Tumor necrosis factor-α(TNF-α) Not determined. Not determined. Not determined. X9 phage display library The values of the dissociation constant KD (μM) were fitted using Nanotemper Analysis software v.2.3 and shown. A lead peptide, pep2 (ACHAWAPTR, KD = 5.14 μM), could directly bind to TNF-α and block TNF-α-triggered signaling activation. Peptide pep2 inhibits TNF-α-induced cytotoxicity and attenuates the inflammation by decreasing NF-κB and MAPK signaling activities in a variety of cells. Furthermore, pep2 attenuated colitis induced by dextran sodium sulfate in mice in both prophylactic and therapeutic settings. Moreover, pep2 reduced the phosphorylation of p38, ERK1/2, JNK1/2, p65, and IκBα in colonic tissues as well as downregulated inflammatory genes. And HIS3, TRP5, and ARG9 may be the key amino acids in pep2 to bind TNF-α by molecular docking. Collectively, targeting TNF-α with pep2 can attenuate the inflammation in vivo and vitro by inhibiting NF-κB and MAPK signaling pathways. 3617 TLWPFDLWLKTR(9/16)[0.2638±0.1027] SIWPFFNSFMTM(3/16)[0.2347±0.0682] TFWNWLPISRAI(2/16)[0.2767±0.0998] TFSWWPIPLPWG(1/16)[0.2593±0.058] TFLPFHWNLSWW(1/16)[0.2825±0.0521] 5 Phage display (subtractive panning) 4 PMID:35066280 Synthetic standard tyrosine-nitrated peptide (sTNP, sequence: H2N-GGGGY*GGG-COOH) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding was quantified by OD450 measurement. OD450 was reproduced from Figure 2B and shown. The whole screening procedure is illustrated in Fig. 1. The phage-displayed 12-mer randomized peptide library was resuspended with TBS supplemented with 1% BSA (w/v) to make a 100 μL phage solution with a tilter of 1 × 1011 pfu/mL, which was incubated with the sepharose 4B immobilized non-nitrated peptides GGGGYGGG at 37 °C for 1 h. The supernatant was collected and further incubated with sepharose 4B immobilized standard TNP (sTNP, sequence: H2N-GGGGY*GGG-COOH) at 37 °C for 1 h. Then, the collected beads were washed five times with 1 mL TBST (TBS supplemented with 0.2% Tween-20) to clean the unbound phages and the bound phages were eluted with 1 mL elution buffer (0.2 M Gly-HCl supplemented with 1% BSA, pH = 2.2). The eluent was immediately neutralized by adding 0.15 mL neutralization buffer (1 M Tris-HCl, pH = 9.1). Collected phages were amplified and titer determination was performed following the manufacturer's instructions. A total of four rounds of the aforementioned “screening-amplification-titer determination-screening” cycle were conducted. Phages from the last biopanning were randomly selected and amplified for ELISA analysis to confirm their binding with the target peptide. Note a 10 or 100 folds decreased amount of target peptide was used for every next round of biopanning to increase the specificity of the screening. SPR analysis suggested that NT-1 (H2N-TLWPFDLWLKTR-COOH) should bind specifically to the nitrated tyrosine. Most importantly, immobilized NT-1 efficiently captured various types of endogenous TNPs in the presence of 100–1000 folds excessive amount of trypsinized BSA fragments. Compared with the commercially used anti-nitrotyrosine antibody, NT-1 enriched more TNPs with less sequence preference in the same experimental conditions. In summary, this study provided NT-1 as a useful start point to design and develop high-affinity binding ligand for TNPs, which could be useful in identifying nitrated tyrosine in proteins in physiological and pathophysiological conditions. 3618 GAQTCMN(15) 1 Phage display (common panning) 4 PMID:36353454 Mycolic acid Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Absorbance was measured at 450 nm and reproduced from Figure 1D and shown. The efficacy of APTX4870 (GAQTCMN) against mycolic acid was demonstrated by evaluating clinical samples and conducting in vitro and Vivo. APTX4870 inhibited apoptosis, increased autophagy to decrease inflammation, and reduced Mycobacterium-derived mycolic acid (M.tb-MA)-induced lung damage. These findings suggest that this heptapeptide, which selectively targets M.tb-MA, might be exploited as a potential novel M.tb therapeutic treatment. 3619 CRGAMWMYKRC(5/6) CEGLAIQVKQC(1/6) 2 Phage display (competitive panning) 3 PMID:36070465 Hyaluronan-binding protein 2 Not determined. Not determined. Not determined. CX9C phage display library Tubes and streptavidin-coated Dynabeads (Thermo Fischer Scientific, Oslo, Norway) were blocked with PBS with 4% (w/v) skim milk powder. Approximately 1 × 10^13 virions of library and 1−10 μg (R1, 10 μg; R2, 3:1 μg) of biotinylated pro-FSAP were incubated with the appropriate amount of Dynabeads for 60 min at room temperature (RT). The beads/ protein/phage complexes were separated from the supernatant using a magnetic rack and washed with PBS-T20 (0.1% (v/v) Tween-20 in phosphate-buffered saline (PBS)). The complexes were then incubated with an excess amount of unbiotinylated pro-FSAP for 60 min at RT to remove phages bound with low-affinity and retain only high-affinity binding clones. The beads/protein/phage complexes were then separated from the supernatant containing the soluble FSAP using a magnet rack and washed with PBS-T20 (0.1% (v/v) Tween-20 in PBS) and with PBS pH 7.4. This was applied to both libraries and in all rounds. The remaining phages were then eluted from the beads with 500 μL of 100 mM triethylamine (pH 11) and neutralized in Tris−HCl pH 7.4. Using a phage display approach, we have identified a Cys-constrained 11 amino acid peptide (CRGAMWMYKRC), NNKC9/41, that activates pro-factor VII Activating protease (FSAP) in plasma. The synthetic linear peptide has a propensity to cyclize through the terminal Cys groups, of which the antiparallel cyclic dimer, but not the monocyclic peptide, is the active component. Other commonly found zymogens in the plasma, related to the hemostasis system, were not activated. Binding studies with FSAP domain deletion mutants indicate that the N-terminus of FSAP is the key interaction site of this peptide. In a monoclonal antibody screen, we identified MA-FSAP-38C7 that prevented the activation of pro-FSAP by the peptide. This antibody bound to the LESLDP sequence (amino acids 30−35) in an intrinsically disordered stretch in the N-terminus of FSAP. The plasma clotting time was shortened by NNKC9/41, and this was reversed by MA-FSAP-38C7, demonstrating the utility of this peptide. Peptide NNKC9/41 will be useful as a tool to delineate the molecular mechanism of activation of pro-FSAP, elucidate its biological role, and provide a starting point for the pharmacological manipulation of FSAP activity. 3620 IDCLMQNAGSA(10/11) DLPWSMPRPCR(1/11) 2 Phage display (competitive panning) 3 PMID:36070465 Hyaluronan-binding protein 2 Not determined. Not determined. Not determined. X11 phage display library Tubes and streptavidin-coated Dynabeads (Thermo Fischer Scientific, Oslo, Norway) were blocked with PBS with 4% (w/v) skim milk powder. Approximately 1 × 10^13 virions of library and 1−10 μg (R1, 10 μg; R2, 3:1 μg) of biotinylated pro-FSAP were incubated with the appropriate amount of Dynabeads for 60 min at room temperature (RT). The beads/ protein/phage complexes were separated from the supernatant using a magnetic rack and washed with PBS-T20 (0.1% (v/v) Tween-20 in phosphate-buffered saline (PBS)). The complexes were then incubated with an excess amount of unbiotinylated pro-FSAP for 60 min at RT to remove phages bound with low-affinity and retain only high-affinity binding clones. The beads/protein/phage complexes were then separated from the supernatant containing the soluble FSAP using a magnet rack and washed with PBS-T20 (0.1% (v/v) Tween-20 in PBS) and with PBS pH 7.4. This was applied to both libraries and in all rounds. The remaining phages were then eluted from the beads with 500 μL of 100 mM triethylamine (pH 11) and neutralized in Tris−HCl pH 7.4. 3621 IMPIQMRRMQML(24)[0.6163±0.0085] IKRIMRPIRQSI(2)[0.4193±0.0125] MRISRPIMRQIT(2)[0.4088±0.0146] RRSHSPMRMPRK(1)[0.4172±0.0105] 4 Phage display (common panning) 3 PMID:33410516 Anti-hemagglutinin (HA) monoclonal antibody PR8-23 Hemagglutinin, HA Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance at 450 nm was measured. Data shown were reproduced from Figure 3B. Biopanning was performed according to the instructions of the manufacturer of the 12-mer peptide phage display library (New England Biolabs), with modifications. Three rounds of biopanning were conducted for antibody PR8-23. The biopanning in the three rounds was identical, except for the increased concentration of Tween-20 (0.5% [vol/vol]) used in the buffer solution in the second and third rounds and the reduction of the concentration of antibody PR8-23 by half in the third round. Ninety-six-well plates were coated with antibody PR8-23 (100 μg/ml) overnight at 4°C. The next day, the plates were blocked with 5% bovine serum albumin (BSA; Sigma-Aldrich) for 1 h at 4°C and then subjected to six washes with buffer solution (50 mM Tris-HCl, 150 mM NaCl, pH 7.5; containing 0.1% [vol/vol] Tween-20). Then, peptide phage library was added (1.62 × 10^11 in 100 μl/well) and incubated at room temperature for 30 min, which was followed by 10 washes with the same buffer solution. Subsequently, the elution buffer (0.2M glycine-HCl, pH 2.2) was used to wash the bound phages at room temperature for 30 min, and then the solution was neutralized to pH 7.8 with Tris-HCl (1M, pH 9.1). Finally, the Escherichia coli strain ER2738 was used to amplify the bound phages, and polyethylene glycol/NaCl (20% [wt/vol] polyethylene glycol-8000, 2.5M NaCl) was used for extraction. Clone 3 showed the highest reactivity among the four phages compared with the control antibody in phage ELISA. And Clone 3 displayed peptides with IMPIQMRRMQML, which interacted with 63‐IAPLQLGKCNIA‐74 (63aa‐74aa) on the HA protein of the A/PuertoRico/8/34(H1N1). 3622 CNLNTIDTC(1/70)[253 ± 79] CNEWQLKSC(1/70)[85 ± 18] CAQRPMKRC(1/70)[NT] CMDVNTFSC(1/70)[NT] CHELSHESC(1/70)[NT] CNDTTFTMC(1/70)[NT] CSKLTKSKC(1/70)[NT] CYNAYMNTC(1/70)[NT] CRGVKTNRC(1/70)[NT] CLRNVTQKC(1/70)[NT] CKLMMPADC(1/70)[NT] CVNMGGIPC(1/70)[NT] CEEHCINVC(1/70)[NT] CSDMMPIDC(1/70)[NT] CIQKSITKC(1/70)[NT] CIAPRKSKC(1/70)[NT] CKLSKKTTC(1/70)[NT] 17 Phage display (subtractive panning) 3-5 PMID:33391537 CD44 variant 6 (CD44v6) overexpressing HEK 293 cells Not determined. Not determined. Not determined. Byungheon Lee (Department of Biochemistry and Cell Biology, School Medicine, Kyungpook National University, Republic of Korea) Surface plasmon resonance (SPR) The binding affinity (KD) of CD44v6-binding peptides to the CD44v6 protein was measured using an SPR instrument (Reichert Technologies, Depew, NY). KD values (nM) were calculated by analyzing the steady-state binding data by fitting the curve of binding level against concentration with a 1:1 binding model (GraphPad Prism 7.0 software). The T7 415-1b phage vector was purchased from Novagen (Madison, WI) and used to construct a phage library displaying CX7C (C, cysteine; X7, seven random amino acids in which hydrophobic amino acids were enriched to occur at a frequency of at least one per every seven residues). The library had a diversity of approximately 1 × 10^9 plaque-forming units (pfu)/mL. Phages (1 × 10^9 pfu) were then incubated with CD44v6 expression vector-transfected HEK 293 cells at 4 °C for 1 h. Phages bound to the cells were eluted by incubation with 500 µL of a suspension of BL21 host bacteria for 10 min at room temperature. To remove phages that had bound non-specifically to cells, the eluted phages were then incubated with non-transfected parental HEK 293T cells at 4 °C for 30 min. Unbound phages in the supernatant were collected, diluted in 10 mL of LB broth, and amplified by culture with BL21 host bacteria. The amplified phages were used in the next round of the screening cycle. A total of 60 phage clones were selected after the third, fourth, and fifth rounds of screening with the transfected cells, and 10 phage clones were selected after the fifth round of screening with non-transfected cells. DNA inserts in the selected phage clones were subjected to a sequencing analysis by Macrogen Inc. (Seoul, Korea). The corresponding amino acid sequences were aligned and analyzed using the Clustal W program to identify shared amino acid sequences or consensus motifs. CNLNTIDTC (NLN) and CNEWQLKSC (NEW) peptides bound preferentially to CD44v6-high cells than to CD44v6-low cells. The binding affinities of NLN and NEW to CD44v6 protein were 253 ± 79 and 85 ± 18 nM, respectively. Peptide binding to CD44v6-high cells was inhibited by the knockdown of CD44v6 gene expression and competition with an anti-CD44v6 antibody. A pull-down assay with biotin-labeled peptides enriched CD44v6 from cell lysates. NLN and NEW induced CD44v6 internalization and inhibited hepatocyte growth factor-induced c-Met internalization, c-Met and Erk phosphorylation, and cell migration and invasion. In mice harboring tumors, intravenously administered NLN and NEW homed to the tumors and inhibited metastasis to the lungs. When combined with crizotinib, a c-Met inhibitor, treatment with each peptide inhibited metastatic growth more efficiently than each peptide or crizotinib alone. In addition, KLAKLAKKLAKLAK pro-apoptotic peptide guided by NLN (NLN-KLA) or NEW (NEW-KLA) killed tumor cells and inhibited tumor growth and metastasis. No significant systemic side effects were observed after treatments. 3623 CWTMVPWGW(4) CSEWWNWWTC(4) CEWYFGWWDC(2) CGPIWGLAG(2) CWWHWNSLW(2) CVWSFWGMY(2) CEIWWGVWLC(2) CWMQWWSPWC(2) CSGWFWAPW(2) CDYWWSNWYC(1) CYWWLPWSAC(1) CWPIMYWWEC(1) CSIYWWEWWC(1) CTYWQYFVWC(1) CKLFFRWIDC(1) CFPFFSQLVC(1) CEWFFGPLVC(1) CAGFFGWVVC(1) CSDFWRWLWC(1) CVHSAFGPWC(1) CWGSPFGWWC(1) CRWAWRWMWC(1) CAWYWGWVEC(1) CVRTWISMVC(1) CEWSWVWDWC(1) CRGVIWSWVC(1) CRGFWPSWVC(1) CGMWWPWVVC(1) CRCFGLRFC(1) CPWCFFGGRC(1) CPWWLFGYPC(1) CAFWIGGLWC(1) CNHAWAAGSC(1) CAWWLTVGLC(1) CHAWWLGGDC(1) CHSWWLGMWC(1) CLWWWEMGGC(1) CSWPVLWWYC(1) CQWLLFSSGC(1) CIDGWFMSAC(1) CLVLWMSAGC(1) CGLFLWRHSC(1) CWKLWRGSRC(1) CVRATSLMGC(1) CVKGRSLFPC(1) CLGLSWFGNC(1) CLSGIAVFLC(1) CGVDVFLWGC(1) CSLLGDVFFC(1) CGWWQWPYLC(1) CHWSSLLRAC(1) CGVGIMWWL(1) CVYWPDWPW(1) CYFWPWEDE(1) CATWGWPWWC(1) CLWPWWFNDC(1) CVWLWWEPYC(1) CMWTWWGSFC(1) CFWSWWGADC(1) CMQQWMGFPC(1) CDLIWWWSDC(1) CGRLAGGLTC(1) CWAIGYGPWC(1) CMLGFLMPGC(1) CWMEWFRWDC(1) CWWWDAWFLC(1) CSGLYKWWLC(1) CLFATGWFLC(1) CAWPWAGWWC(1) CGWFWGGWWC(1) CWLAGWKVLC(1) CNYGWSILC(1) CRVWPGLLMC(1) CPGWLPRLC(1) CEWWYWALGC(1) 75 Phage display (subtractive panning) 4 PMID:34643765 Chemically synthesized crotamine, SCRO Not determined. Not determined. Not determined. CX7C and CX8C phage display library pool ELISA For peptide binding validation, SCRO-coated plates were blocked with 3% casein in PBS and then incubated with phage diluted in Tris-buffered saline (TBS). After washing, rabbit anti-Fd bacteriophage antibody (Sigma, St. Louis, MO, USA Cat# B7786) was added, followed by anti-rabbit IgG conjugated to horseradish peroxidase (Santa Cruz, Dallas, TX, USA). The ELISA was developed with 3,3′,5,5′-tetramethylbenzidine.  The screen for crotamine-binding peptides was performed using a 50:50 mixture of phage cyclic peptide libraries CX7C and CX8C (C: cysteine; X: any amino acid residue) provided by Erkki Koivunen. Maxisorp plates (Nunc, Thermo, Rockford, IL, USA) coated with SCRO were exposed to the phage library that had previously been depleted of non-specific phages by exposure to bovine serum albumin (BSA)–treated plates. After incubation, plates were washed and selected phages were amplified through K91 E. coli infection and quantified by colony counting. The viruses purified from these bacteria were named “enriched round 1.” Phage panning was repeated with the enriched library for another three rounds. Round 4 bacterial colonies were used for sequencing. One of the peptides (CVWSFWGMYC), synthesized chemically, was shown to bind both synthetic and natural crotamine and to block crotamine-DNA binding. 3624 VLGREEWSTSYW[10.6105] LEKGNTLSTSTV[NT] NPIVRSAEDGQL[NT] NESGITRIALQD[NT] YDADMFYMKTNM[NT] 5 Phage display (common panning) 3 PMID:36456600 Extracellular domain of epidermal growth factor tyrosine kinase receptor mutation variant III (EGFRvIII) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Flow Cytometry Ph.D.™-12 Phage Display Peptide Library Kit was purchased from New England Biolabs Inc. (Beverly, MA, USA). Biopanning procedures were done according to the manufacturer’s instruction with certain modifications. Briefly, a 96-well plate was coated with 150 µL hEGFRvIII (Sino Biological #29662-H08B) (in the first two rounds at 100 μg/mL, the third round at 10 μg/mL) overnight at 4 °C. Wells were washed with PBS, blocked with blocking buffer (3% BSA in PBS), washed six times with cold PBST (PBS + 0.1% [v/v] Tween-20), then incubated with 1010 pfu phage peptide library Ph.D.™-12 for 2 h at RT. Unbound phages were removed by washing 10 times with cold PBST (PBS + 0.1% [v/v] Tween-20 in the first round and 0.5% [v/v] in the other two rounds). Bound phages were eluted with 100 μL of 0.2 M Glycine–HCl (pH 2.2), 1 mg/mL BSA and neutralized with 15 μL of 1 M Tris–HCl, pH 9.1. The elution procedure was repeated three times and the final eluate was used for amplification in Escherichia coli ER2738 culture. Recovered phages were subjected for two more rounds of biopanning with hEGFRvIII proteins (Sino Biological #29662-H08B). The eluates of each round were titrated by qPCR to enumerate phages and eluate from the third round of screening for NGS sequencing. Streptavidin was used as a biopanning control with the same condition as the target except for elution. The unbound phages were eluted with 100 μL of 0.1 M Biotin in PBS for 30 min. The enriched peptides were characterized and their binding capacity towards stable cell lines expressing cancer-specific epidermal growth factor tyrosine kinase receptor mutation variant III (EGFRvIII), EGFR wild type (EGFR WT), or a low endogenous level of EGFR WT was confirmed by flow cytometry analysis. The best peptide candidate, VLGREEWSTSYW, was synthesized, and its binding specificity towards EGFRvIII was validated in vitro. Additionally, computational docking analysis suggested that the identified peptide binds selectively to EGFRvIII. The novel VLGREEWSTSYW peptide is thus a promising EGFRvIII-targeting agent for future applications in cancer diagnosis and therapy. 3625 CAINSLSRKC(21%) CAKSMGDIVC(11%) CGRKQVESSC(10%) CRGKSAEGTC(5%) 4 Phage display (in vivo) 4 PMID:33870243 Integrin alpha-3 Not determined. Not determined. Not determined. fUSE5-based CX8C phage display library Phage was 10^9 TU of the phage library per mouse administered via the intratracheal route with 50 μL of PBS with a MicroSprayer® Aerosolizer coupled to a high-pressure syringe (Penn-Century) and a small animal laryngoscope (Penn-Century). The devices were used to administer air-free liquid aerosol directly into the trachea of animals deeply anesthetized with 1% isoflurane. The four rounds of selection were performed as described. In round one (R1), animals received 109 TU of the CX8C library via aerosol (see scheme in Figure 1B). After 60 min, phage particles were recovered from the bloodstream, amplified and pooled. In round two (R2), R1-pooled phage particles (i.e., inhaled phage sub-library 1) were administered and recovered 30 min post-administration. The subsequent R2-pooled phage particles (i.e., inhaled phage sub-library 2) were amplified for administration in round three (R3). After 10 min, the R3-pooled phage particles (i.e., inhaled phage sub-library 3) were recovered and processed for aerosol administration in the final round four (R4). After 5 min, phage particles were recovered from the bloodstream, amplified, and the corresponding peptide-encoding genomes were analyzed by DNA sequencing. We screened a phage display random peptide library in vivo to select, identify, and validate a ligand (CAKSMGDIVC) that specifically targets and is internalized through its receptor, α3β1 integrin, on the surface of cells lining the lung airways and alveoli and mediates CAKSMGDIVC-displaying phage binding and systemic delivery without compromising lung homeostasis. As a proof-of-concept, we show that the pulmonary delivery of targeted CAKSMGDIVC-displaying phage particles in mice and non-human primates elicit a systemic and specific humoral response. 3626 TDFLRMMLQEER(8/16) 1 Phage display (common panning) 3 PMID:35534543 Anti-interferon gamma (IFN-gamma) monoclonal antibody B27 Interferon gamma, IFN-gamma Not determined. Not determined. SUT12 M13 phage display library ELISA Their binding activity against B27 mAb was detected by phage ELISA. Data were not shown. The specific peptide sequence recognized by B27 mAb, TDFLRMMLQEER, was retrieved from a phage display random peptide library. Sequence alignment and homology modeling demonstrated that the queried phage peptide sequence and structure were similar to amino acids at position 27–40 (TLFLGILKNWKEES) of the human IFN-γ. 3627 DYHDPSLPTLRK(4)[0.7929±0.1464, NT] GNNPLHVHHDKR(2)[0.6135±0.0746, NT] STVRPLLMMDKY(1)[1.2004±0.2109, NT] HHLRIPYALDQT(1)[0.5489±0.0904, NT] DSAPSYNYRPSY(1)[1.6395±0.1421, 520±7] DYHDPSPPTLRK(1)[0.6436±0.2095, NT] KVYFSIPWRVPM(1)[1.6281±0.198, 74±3] QVNGLGERSQQM(1)[1.1559±0.1119, NT] HSNDPRLITMRK(1)[0.3179±0.2239, n.d.] TCFAHTHNNFGH(1)[0.8001±0.3975, NT] RDYHPRDHTATW(1)[0.3394±0.1579, NT] IPGTAPPLARTG(1)[1.044±0.023, NT] KDFLPSPQTATW(1)[1.5606±0.1277, 488±33] VRAFSGEHSFVS(1)[1.1746±0.023, NT] 14 Phage display (common panning) 3 PMID:35059169 9-fluorenylmethoxycarbonyl-phenylalanine-tyrosinephosphate (Fmoc-Fργ ) micellar aggregates Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance at 405 nm was monitored in a SpectraMax i3 (Molecular Devices). Wild-type M13 phage has been used as a positive control of ELISA assay and as a negative control of Fmoc-FpY binding. In addition, NMR experiments were acquired in a Bruker Avance III 600 spectrometer equipped with a triple-resonance cryoprobe (TCI). Spectra were processed using software TOPSPIN (Bruker Biospin, Karlsruhe, Germany) and peaks assigned with Sparky (TD Goddard and DG Kneller, Sparky 3, University of California, San Francisco, USA). The dissociation constants KD (μM) were determined in the OriginPro9. The first column of affinity values is the absorbance405 value reproduced from Figure 1C, the second column is the KD (μM) determined by NMR experiments. The lead peptide, KVYFSIPWRVPM-NH2 (P7) was found to bind to the Fmoc-FpY ligand exclusively in its self-assembled state with KD =74 ± 3 μM. Circular dichroism, NMR and molecular dynamics simulations revealed that the peptide interacts with Fmoc-FpY through the KVYF terminus and this binding event disrupts the assembled structure. In absence of the target micellar aggregate, P7 was further found to dynamically alternate between multiple conformations, with a preferred hairpin-like conformation that was shown to contribute to supramolecular ligand binding. Three identified phages presented appreciable binding, and two showed to catalyze the hydrolysis of a model para-nitro phenol phosphate substrate, with P7 demonstrating conformation-dependent activity with a modest kcat/KM = 4 ± 0.3 * 10^(-4) M^(-1)s^(-1). 3628 ACVWWDHTYC(8/19)[2.0156±0.0932] ACNWWDLTLC(6/19)[1.8097±0.0919] ACTWWDMAFC(4/19)[1.8452±0.1287] ACEWWDVTYC(1/19)[2.5513±0.0711] 4 Phage display (subtractive panning) 3 PMID:37126866 Anti-CPA monoclonal antibody MAB-1418 Cyclopiazonic acid, CPA Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The analytical signal was obtained by measuring the absorbance at 450 nm with a CLARIOstar microplate reader from BMG (Ortenberg, Germany). Data shown were reproduced from Figure 2. A pre-selection step against BSA was carried out to eliminate non-specific phages prior to the actual selection with mAb-1418 in each round. The mimotope sequence, ACNWWDLTLC, performed best in phage-based immunoassays, surface plasmon resonance binding analyses, and UC-RET-based immunoassays. To develop a homogeneous assay, upconversion nanoparticles (UCNP, type NaYF4:Yb3+, Er3+) were used as energy donors and coated with streptavidin to anchor the synthetic biotinylated mimotope. Alexa Fluor 555, used as an energy acceptor, was conjugated to the anti-CPA antibody fragment. The homogeneous single-step immunoassay could detect cyclopiazonic acid (CPA) in just 5 min and enabled a limit of detection (LOD) of 30 pg/mL (1.5 μg/kg) and an IC50 value of 0.36 ng/mL. No significant cross-reactivity was observed with other co-produced mycotoxins. Finally, we applied the novel method for the detection of CPA in spiked maize samples using high-performance liquid chromatography coupled to a diode array detector (HPLC-DAD) as a reference method. 3629 AGKCCFATGGS(5) ARKCCYGRGGS(4) ASRKCCYVGGS(2) AGGKCCYSGGS(1) AGKCCYSKGGS(1) AGKCCYSTGGS(1) AKCCYAGRGGS(1) AKCCYAHIGGS(1) ANRKCCFAGGS(1) ARAKCCYYGGS(1) ASKCCYHVGGS(1) AYKCCYWTGGS(1) ASSFLEQDGGS(1) AILIPFWTGGS(1) AIPYCMGVGGS(1) AWLRSMKVGGS(1) ANTGWQSRGGS(1) AVFFGVGWGGS(1) AKKVWIIMGGS(1) AMMTSSGGGGS(1) AYFVVRGLGGS(1) KCC[FY] 21 Phage display (common panning) 3-4 PMID:35665749 Zc RNase P ribozyme (RPR) Not determined. Not determined. Not determined. AXXXXXXXGGS phage display library We report that a hydrophobic-cationic RNA binding peptide selected by phage display (P43: AKKVWIIMGGS) forms insoluble amyloid-containing aggregates, which reversibly accrete RNA on their surfaces in an RNA-length and Mg2+- concentration dependent manner. The aggregates formed by P43 or its sequence-simplified version (K2V6: KKVVVVVV) inhibited RNA polymerase ribozyme (RPR) activity at 25mM MgCl2, while enhancing it significantly at 400mM MgCl2. Our work shows that such hydrophobic-cationic peptide aggregates can reversibly concentrate RNA and enhance the RPR activity, and suggests that they could have aided the emergence and evolution of longer and functional RNAs in the fluctuating environments of the prebiotic earth. 3630 RRPPR(5/24) 1 Phage display (common panning) 6 PMID:34111267 Rat heart microvascular endothelial cell, RHMVEC Not determined. Not determined. Not determined. Novagen T7 X7 phage display library Rat heart microvascular endothelial cell (RHMVEC) (80% confluent; approximately 2.0e7 cells / 100 mm dish) were washed with PBS and preincubated in serum-free medium at 37°C for 30 minutes and inoculated with an extract (5.0e9 pfu) of the T7 phage library to reach a multiplicity of infection (MOI) of 250. After incubation for 1 hour at 37°C, cells were washed with ice-cold PBS and acid washed with 0.1N HCl pH 2.2 for 15 seconds to remove unbounded and weakly associated phages from the cell surface. Cells were then trypsinized, centrifuged, and lysed with sterile deionized water on ice and internalized phages were amplified. After completion of 6 rounds of selection/amplification, Escherichia Coli BL21 was infected with the resulting phages and plated, individual plaques were picked, amplified, and sequenced. Phage display technology was used to identify a small peptide (RRPPR) that was internalized into endothelial cells. This peptide was potent in blocking nitric oxide (NO) release. Fusing RRPPR with a minimal Cav inhibitory domain (CVX51401) dose-dependently blocked NO release, VEGF induced permeability, and retinal damage in a model of uveitis. 3631 AYPGLPRSQS[155] AYPGPRTGHW[inactive] AYPGPHTFRG[inactive] AYPGWTTKVA[53] AYPGWPLRNA[61] AYPGWLVKNG[4.8] 6 Phage display (competitive panning) 3 PMID:35343875 Membranes of HEK293 cells expressing protease-activated receptor 4 (PAR4), HEK293-PAR4 Not determined. Not determined. Not determined. AYPGX6 phage display library Changes in fluorescence intensity were measured using FLIPR Tetra Systems. EC50 (μM) from the individual fits were calculated and shown. To obtain binding peptides by phage display, we opted to pan with membranes derived from HEK293 cells transiently expressing PAR4 receptor (HEK293-PAR4). The first panning cycle was carried out by incubating an approximately 10^10 library phage in a well coated with HEK293-PAR4 membranes. After 5 rinses and 1 x 5-minute wash with PBST (phosphate buffer saline containing 0.1% Tween 20), binding phage were then recovered and amplified by infection with log phage E. coli strain ER2738 (New England Biolabs). The second round of panning was carried out similarly to the first cycle, except that: (1) steps were carried out in duplicate wells so that binding phage could be recovered from the first well by infection and from the second well by elution with 200 μM AYPGKF peptide; (2) the panning step was preceded by 3 successive 30-minute pre-adsorptions in wells coated with wild-type HEK293 membranes; (3) amplified output phages from the first round were mixed with 0.2 μg of αVβ3 integrin (Chemicon, Burlington, MA) so that a competitive depletion of integrin-binding phage was possible in the panning step; and (4) wash steps included 5 fast rinses, 2 x 5-minute washes, and a further fast rinse. The third panning cycle was as cycle 2 except that: (1) 5 x 15-minute pre-adsorptions with HEK293 membranes were carried out; and (2) elution with ER2738 was replaced by a 10-minute elution with 0.1 M glycine buffer, pH 2.2, followed by neutralization with 0.1 vol of 2M Tris, pH 8.0. Throughout the steps, membranes were immobilized for 2 hours in wells of 96-well plates coated with wheat-germ agglutinin (Perkin Elmer, Waltham, MA) and blocked for 1.5 hours with PBS plus milk (5% fat free), followed by 4 rinses with PBS. All phage incubations were performed for 2 x 2 hours in PBS plus milk. All steps were carried out at 4℃ in the presence of a protease inhibitor cocktail. Using an AYPG-based biased phage-display peptide library approach followed by chemical peptide optimization, A-Phe(4-F)-PGWLVKNG was identified. This peptide demonstrated an EC50 value of 3.4 μM in a platelet-aggregation assay, which is 16-fold more potent than AYPGKF. Using this new protease-activated receptor 4 (PAR4) agonist peptide (AP), a platelet-rich plasma-aggregation assay using light-transmission aggregometry was developed and validated in a series of precision and reproducibility tests. PAR4 antagonist responses to PAR4 AP A-Phe(4-F)-PGWLVKNG (12.5 μM to 100 μM) were subsequently evaluated in this assay in vitro and ex vivo in a human study using BMS-986120, a PAR4 antagonist that entered clinical studies. 3632 GSYWYNVWF 0 Phage display (common panning) PMID:34079028 IgG Fc region human MDA-MB-231 cells and mouse EF43.fgf4 cells Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA peptide ligand binding to human IgGs 3633 NGFFEPWQVVYV 1 Phage display (competitive panning) 3 PMID:35007993 Maltose-binding protein (MBP) (MBP-VHH2,4-D) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance at 450 nm were measured by SpectraMax M5. The bound phages were competitively eluted by a 2,4-D standard solution and followed added to the wells only MBP coated to further remove undesired phages. A variable domain (VHH) to 2,4-dichlorophenoxyacetic acid (2,4-D) was used to isolated the peptidomimetic (NGFFEPWQVVYV) from phage display libraries using six panning conditions. Then the peptidomimetic and VHH were fused with the larger (LgN) and smaller piece (SmN) of split fragments of Nluc, respectively. In order to optimize the signal and sensitivity of the immunosensor, we explored the effects of the spacer between the peptidomimetic and LgN, the copy number of peptidomimetics, and the spacer between SmN and VHH, generating 24 combinations that allowed to conclude on their respective roles. Eventually, the developed “ready-to-use” immunosensor performed excellent signal-to-noise ratio and sensitivity, and could be applied to the detection of 2,4-D in real samples. 3634 SRHGQRALQALP(8/15)[1.1428±0.0442] 1 Phage display (common panning) 4 PMID:33567652 Anti-canine adenovirus 2 (CAdV-2) monoclonal antibody 2C1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The analytical signal was obtained by measuring the absorbance at 450 nm. Data shown were reproduced from Figure 3a. The peptide, SRHGQRALQALP, is similar to the sequence "RIKQRETPAL" observed in the CAdV-2 Hex. 3635 SAPHSASDPHYS(9/15) SESAGGNHYSHP(2/15) FYQHSNPWAKYS(1/15) PLSRYKTRAYSC(1/15) VPESRYTVPSAT(1/15) PESXXDXXYS 5 Phage display (common panning) 4 PMID:33567652 Anti-canine adenovirus 2 (CAdV-2) monoclonal antibody 2C1 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The analytical signal was obtained by measuring the absorbance at 450 nm. Data shown were reproduced from Figure 3b. CAdV-2-specific neutralizing monoclonal antibodies The high affinity phage clones with mAb 7D7 displayed a consensus motif PESXXDXXYS (X is any aa), similar to the sequence "PESYKDRMYS" observed in the CAdV-2 Hex. 3636 CIHSPTALC(5124) CNAGHLSQC(4414) CNIKSSHVC(4296) CNMHTPMVC(4232) CISSSINHC(3804) CTLKNLALC(3787) CLTAKHMQC(3715) CLAYAHHTC(3457) CNWMINKEC(3267) CFATRADHC(3089) CDGLAKNSC(3028) CPKGDENTC(2982) CSAQQPASC(2966) CNDTKQGNC(2904) CGPTAKYIC(2867) CPSTVPWSC(2856) CVPSKPGLC(2826) CNSHTQGKC(2522) CSVGYDRNC(2487) CSENSPLLC(2478) CSKEATPFC(2445) CLDHSSKLC(2182) CNTSTMLHC(2181) CEQAHKLHC(2172) CTTKLPNSC(2066) CNANPWRLC(2054) CLNRYVADC(2027) 27 Phage display (common panning) PMID:35871689 Escherichia coli MSI001 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) A representative heptapeptide CVTKLGSLC (VTK) was selected for fusion with the antibacterial peptide LL-37 to construct the specific-targeting antibacterial peptide VTK-LL37. We found that, in comparison with LL37, VTK-LL37 showed prominent bacteriostatic activity and an inhibitive effect on biofilm formation in vitro. In vivo experiments demonstrated that VTK-LL37 significantly inhibited bacterial growth, reduced HMGB1 expression, alleviated lesions of vital organs and improved the survival of mice subjected to CLP modeling. Furthermore, membrane DEGP and DEGQ were identified as VTK-binding proteins by proteomic methods. 3637 CVPKRRNNC[0.4316±0.0255] CLKRQRT[0.3551±0.0538] CRRRVS[0.3453±0.0353] CNRRRA[0.5789±0.0212] CRKRS[0.4203±0.027] CRKIR[0.3482±0.0324] 6 Phage display (subtractive panning) 4-5 PMID:35014500 Kidney injury molecule 1, KIM-1 Not determined. Not determined. Not determined. Byungheon Lee (Department of Biochemistry and Cell Biology, School Medicine, Kyungpook National University, Republic of Korea) ELISA Optical density data (450 nm) for each clone is expressed as the mean ± standard deviation (SD) of three independent experiments. Data shown were reproduced from Figure 1. Each biopanning round consisted of subtraction of phages nonspecifically bound to M-270 epoxy beads only and subsequent selection of phages bound to KIM-1 conjugated epoxy beads with 3 M ammonium sulfate, followed by amplification of the eluted phages. This peptide, with the sequence CNRRRA, not only showed a high imaging potential in vitro, allowing a strong detection of kidney injury molecule-1 (KIM-1) expressing cells by microscopy and flow cytometry but also generated a strong kidney-specific signal in live-imaging in vivo experiments in the context of a drug-induced kidney-injury mouse model. 3638 CTKTDVHFC[2.1512±0.0436] CIHSSTRAC[1.9349±0.0576] CMQTQRAHC[1.8843±0.0297] CTNANHYFC[1.1291±0.0279] CTYENHRTC[1.546±0.0645] CDPRHSKFC[0.9914±0.0436] CLAQSHPLC[1.3227±0.0297] 7 Phage display (competitive panning) 4 PMID:34635758 GTTYGVCSK-biotin and Domain III of the envelope (E) glycoprotein (DIII) Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Optical density data (450 nm) for each clone is expressed as the mean ± standard deviation (SD). Data shown were reproduced from Figure 1B. The first round of panning: target: GTTYGVCSK-biotin (1 μg/well), phage elution: competitive elution with GTTYGVCSK-biotin (9 μg) in PBS (pH 7.2). The second round of panning: target: GTTYGVCSK-biotin (1 μg/well), phage elution: competitive elution with GTTYGVCSK-biotin (2 μg) in PBS (pH 7.2). The third round of panning: target: GTTYGVCSK-biotin (1 μg/well), phage elution: non-related peptide in CovaBuffer (pH 7.2) + 1 M glycine; followed by (ii) competitive elution with GTTYGVCSK-biotin (2 μg) in PBS (pH 7.2). The 4th round of panning: target: rDIII (0.3 μg/well), phage elution: 0.1% TBST (pH7.2) + 250 mM imidazole. Four cyclic peptides (CTKTDVHFC, CIHSSTRAC, CTYENHRTC, and CLAQSHPLC) showed significant blocking of the interaction between DIII and hBMECs, and ability to neutralize infection in cultured cells. 3639 SGVYKVAYDWQH[1.9855±0.0209] HYSWSWIAYSPG[3.0371±0.0139] 2 Phage display (competitive panning) 5 PMID:34635758 GTTYGVCSK-biotin and Domain III of the envelope (E) glycoprotein (DIII) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Optical density data (450 nm) for each clone is expressed as the mean ± standard deviation (SD). Data shown were reproduced from Figure 1B. The first round of panning: target: GTTYGVCSK-biotin (1 μg/well), phage elution: competitive elution with GTTYGVCSK-biotin (9 μg) in PBS (pH 7.2). The second round of panning: target: GTTYGVCSK-biotin (1 μg/well), phage elution: competitive elution with GTTYGVCSK-biotin (2 μg) in PBS (pH 7.2). The third round of panning: target: GTTYGVCSK-biotin (1 μg/well), phage elution: non-related peptide in CovaBuffer (pH 7.2) + 1 M glycine; followed by (ii) competitive elution with GTTYGVCSK-biotin (2 μg) in PBS (pH 7.2). The 4th round of panning: target: rDIII (0.3 μg/well), phage elution: 0.1% TBST (pH7.2) + 250 mM imidazole. The 5th round of panning: target: rDIII (0.3 μg/well), phage elution: competitive elution with GTTYGVCSK-biotin (2 μg) in PBS (pH 7.2). peptides, 4 cyclic peptides showed significant blocking of the interaction between DIII and hBMECs, and ability to neutralize infection in cultured cells. 3640 CSSTRESAC CRYSAARSC CRGFVVGRC CQRALMIAC 4 Phage display (in vivo) 3 PMID:34060472 Mouse mammary EF43.fgf4 cell Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) Binding assay Binding of individual phage clones to EF43.fgf4 cells was quantified by the counting of transducing units (TU) after host bacterial infection. The values of the relative TU were reproduced from Figure 1B and shown. Combinatorial phage display selections in vivo in tumor-bearing mice were performed as follows: animals received 10^9 TU iv of an unselected phage display random peptide library (displaying the insert CX7C). Tumors and control organs were collected after 24 hr of systemic circulation. For homing of individual phage clones in vivo, tumor-bearing mice were deeply anesthetized with 1–2% isofluorane and received 10^9 TU of targeted phage or insertless control phage, both administered iv side-by-side. Phage particles were recovered from tissue samples by bacterial infection and processed. We identified a cyclic peptide (CSSTRESAC) that specifically binds to a vitamin D receptor, protein disulfide-isomerase A3 (PDIA3) expressed on the cell surface of tumor-associated macrophages (TAM), and targets breast cancer in syngeneic triple-negative breast cancer (TNBC), non-TNBC xenograft, and transgenic mouse models. Systemic administration of CSSTRESAC to TNBC-bearing mice shifted the cytokine profile toward an antitumor immune response and delayed tumor growth. Moreover, CSSTRESAC enabled ligand-directed theranostic delivery to tumors and a mathematical model confirmed our experimental findings. 3641 ACSDRFNCPADEALCG(17) KCCNVINCCK(10) KCCTKNCDSTAHCT(9) RCDHCLCLQCS(8) TCCAFCVSCI(7) SCSTCNDSVSDCCL(6) QCMRQPCENCCR(6) TCSRTHQCTRPCCP(5) GCVSCCQGTCF(5) KCCTESPLCCT(3) SCPCEGCVNFSCF(2) SCHDCNTCGW(1) GCCCGSSVCH(1) RCPFPCTPQRCCY(1) RSCWRCSSAPFCCA(1) CVVLHSSEICDCS(1) DCCWQGWCICN(1) MCANCSEDPCCS(1) NCHPCNNCTNCK(1) QCRTQPSCPNCKCK(1) 20 Phage display (common panning) 5 PMID:33976148 Ice Not determined. Not determined. Not determined. XCXmCXnCXoCX phage library pool Our ice-affinity selection protocol enables the selection of a cyclic ice-binding peptide containing just 14 amino acids (KCCTKNCDSTAHCT). Mutational analysis identifies three residues, Asp8, Thr10 and Thr14, which are found to be essential for ice binding. 3642 NRPDSAQFWLHH[3.6] SITNGWKTVIRS[10.2] YIPEVRHSRAWH[16.7] HSSPHFSRHGLL[13.2] QPPLQPIPDALT[1.2] HPDIYFHPGNQR[42.9] GISWQQSHHLVA[34] ATWSHHLSSAGL[31.2] 8 Phage display (common panning) 3 PMID:36701953 Mucin-5B, MUC5B Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Binding assay The amount of peptide in the supernatant of the control and test samples of each peptide was determined using capillary zone electrophoresis. The amount of peptide bound (shown as affinity value) was quantified by comparative analysis of the peak areas of control and test samples in % using BIOFOCUS Integrator software (Bio-Rad). The difference in the areas of peaks of control and test samples of each peptide corresponded to the amount of bound peptide. MBP-12 (HPDIYFHPGNQR) and MBP-14 (GISWQQSHHLVA) displayed the highest affinity to MUC5B. MBP-12 mildly stabilized the spinnbarkeit of serous saliva after overnight incubation and of mucous saliva at all timepoints tested. The addition of MBP-12 to a pellicle of unstimulated saliva on HA discs showed no additive protective effect against acid-induced demineralization. Epitope characterization suggested sulfo-Lewis(a) SO3–3Gal_1–3GlcNAc (galactose residue) as MBP-12 binding site on MUC5B. 3643 TLMVPRTGS(6) WCQVQSVCA(3) RWRRKALPW(2) GWRPKMVLR(2) RWRRKVLRH(2) LSSCFRFCS(1) WNMKFVWRS(1) RPRARVFRF(1) VWREPLHHH(1) ARGPRRGVR(1) RRVLSACVW(1) VTLRFNGWG(1) APAARNLEL(1) VLVPVGVVW(1) GAVSSHLDL(1) GISGVSGCS(1) LVYLVWDVW(1) TWRRRLLVH(1) TWRRRNLVH(1) TWRKRMRLL(1) GWRRKALMR(1) GWRRKVWLR(1) FWRRKVWHH(1) RWRRKVFLH(1) GWRRKVMRL(1) FWRRKVLHH(1) SWRRKIYHL(1) GWRRKLRVV(1) GWRRKSLWA(1) GWRKKSLRL(1) RWRKKAWHH(1) RWKRKVLEA(1) RWKRKVRVV(1) MWKRKVLVL(1) GWKRKTLSL(1) RWKRKSLGR(1) GWKRKYRQV(1) TWKRKWFHH(1) KYKRKRLFL(1) RWKKKSWLW(1) GRYRKNIMH(1) GWFRKMLFS(1) GWRPKTWIG(1) GWKPKWLLD(1) RWRAKVLPV(1) GWRVKSIPV(1) RWRVKSRKW(1) GWRTKVGVL(1) RWRPRVLPH(1) RWRPRNWRH(1) GWRARTLRV(1) RWRARVRRL(1) GWRARVLSL(1) GWRTRVLPL(1) RWRSRVLRI(1) RWRMRALFH(1) GWRPKRGYL(1) SWRPKRLPW(1) GWRAKRVIW(1) GWRAKRMMV(1) GWKAKRLSV(1) RWRVKRYSH(1) GWRVKRLVV(1) GWRLKRLVD(1) RWRVKRAML(1) GWRSKRLRV(1) GFRMKRLVV(1) GWRFKRLEL(1) GWRAKRRII(1) AWRPRRLVV(1) GWRARRLVI(1) GWRVRRLFT(1) GWKARKLVF(1) GWLAKRAHH(1) GWASKRLQV(1) RWFSKRLLH(1) VPRLKILVM(1) ASRLKVHHH(1) VVWGKMLSH(1) WGWLRFEHH(1) PGGWRRKALSV and PGGWRAKVLSV 80 Phage display (common panning) 7 PMID:34919980 Cathepsin G-like Not determined. Not determined. Not determined. X9 T7 phage display library A preference for a lysine in the P1 position of a substrate, arginines in positions P2 and P3, and the aromatic amino acid tryptophane in the P4 position was observed. Based on the sequence alignment we could identify a consensus sequence for this protease as being PGGWRRK↓ALSV. Mass spectrometry analysis of a peptide with the consensus sequence obtained by phage display showed that cleavage of this peptide occurred after the conserved Lys (K) residue. 3644 WDLPPIGRLSGN(50.0%) SWMPPILRSPAV(26.3%) THLPPIMRNLQF(5.7%) SWLPNIQRHWLS(2.6%) HLPPILRMLDLV(2.6%) HLPPIQRTPTYA(2.6%) LPPIVRLPGLLH(2.6%) FPFGPINRDMTA(2.6%) NGVWLPPIARVL(2.6%) LPPIXRX 9 Phage display (common panning) 3 PMID:36312416 Good Vibes phage, GV Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured at 450 nm using an Infinite 200 Rx plate reader. The heat map of Figure 3C shows the cross-reactivity of GV monoclonal phages against GV (target), MAT (same family as GV), cowpea mosaic virus (CPMV, unrelated plant virus), and bovine serum albumin (BSA). We screened a phage display library for peptides that bind to the Good Vibes phage (GV), which lyses the bacterial pathogen Pseudomonas aeruginosa. Isolated monoclonal library phages featured a highly conserved consensus motif, LPPIXRX. The corresponding peptide WDLPPIGRLSGN was synthesized with a GGGSK linker and conjugated to cyanine 5 or biotin. The specific binding of the LPPIXRX motif to GV in vitro was confirmed using an enzyme-linked immunosorbent assay. We demonstrated imaging and tracking of GV in bacterial populations using the fluorescent targeting peptide and flow cytometry. 3645 RWKDTAYALTNN(4)[NB] SWHWHTHVRHQM(3)[0.04 ± 0.01] WGHSHFSHWKGR(3)[6.86 ± 2.63] HHLRIPYALDQT(2)[NB] SHRWQVWSRDRA(2)[NB] ASANDNRLRYTY(2)[NB] LDRPSSLAHLAS(2)[2.14 ± 0.69] RHHSSNPRDTAP(2)[8.82 ± 0.01] 8 Phage display (common panning) 5 PMID:36195696 Cytotoxic T-lymphocyte protein 4 T-lymphocyte activation antigen CD80 Not determined. Not determined. Ph.D.-12 phage display library (X12) The human CTLA4 binding peptides to hCTLA-4 protein affinity was measured by microscale thermophoresis (MST). The dissociation constant (Kd, μM) was determined and shown. The LC4 peptide (WGHSHFSHWKGR) was modified to improve its tumour-targeting ability and reduce peripheral immune system activation, which could block the CTLA-4/CD80 interaction. The LC4 peptide as a result, like other immune checkpoint inhibitors (ICIs), exerts anti-tumour effects by refreshing T cell function, and also activates the peripheral immune system. We used the PLGLAG peptide as a linker at the C-terminal of LC4 to connect with a tumourtargeting peptide RGD to increase the tumour tissue targeting ability, and obtain LC4-PLG-RGD. Further experiments demonstrated that the anti-tumour LC4-PLG-RGD activity was better than LC4 in vivo, and the ability to activate the peripheral immune system was weakened. 3646 ITNAPIKDLTP NSNDFRPGGPET SNKNLDTRILTK 3 Phage display (common panning) 3 PMID:34260404 APTYSW peptide CD81 antigen Not determined. Not determined. Ph.D.-12 phage display library (X12) 3647 LPLSTQH 1 Phage display (common panning) 5 PMID:34673862 Fe3O4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Special M13 with Fe3O4 affinity pIII-peptide (FAP-M13) was biopanned for strongly binding towards bare Fe3O4 with the “hook”-like pIII-peptide (N-LPLSTQH-C). TEM observation confirmed the direct grasp of FAP-M13 on bare Fe3O4, forming the magnetic (FAP-M13)-Fe3O4 virus hydrogel. 3648 CRTLPFHEC(7/80)[5.9 ± 0.2] CEKMVATHC(4/80)[not determined] CRTLPWNQC(3/80)[2.3 ± 0.6] CRTIPFTHC(2/80)[not tested] CRTMEYTSC(2/80)[not tested] CRTLPYHLC(2/80)[2.7 ± 0.2] CRTQPYNQC(1/80)[not tested] 7 Phage display (subtractive panning) 5 PMID:33668971 WD repeat-containing protein 5 Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The fluorescence polarization was measured in Synergy™ Neo2 Multi-Mode Microplate Reader using an excitation filter at 485 nm and an emission filter at 535 nm. The EC50 (μM) value was determined and shown. To remove false-positive target-unrelated peptides (TUPs) which directly bind to Ni-NTA magnetic beads, the library was depleted 2 times by incubating the phage library directly with Ni-NTA magnetic beads before the biopanning against the target in the second and third rounds. The detailed binding interactions were revealed by solving a WDR5-peptide cocrystal structure. 3649 WSLGYTG(8.9%) 1 Phage display (common panning) 3 PMID:35328728 T-cell surface glycoprotein CD4 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) We report the isolation of a fast-propagating white clone (displaying WSLGYTG peptide) identified through screening against a recombinant protein. Sanger sequencing demonstrated that white plaques are not contamination from environmental M13-like phages, but derive from the library itself. Whole genome sequencing revealed that the white color of the plaques results from a large 827-nucleotide genomic deletion. The phenotypic characterization of propagation capacity through plaque count- and NGS-based competitive propagation assay supported the higher propagation rate of Ph-WSLGYTG clone compared with the library. 3650 GLHTSATNLYLH[0.2704±0.0069] DSQFNKYSIATV[0.2676±0.0194] MHPNAGHGSLMR[0.2055±0.0076] DEVPPGMTLPRP[0.2338±0.0262] SGVYKVAYDWQH[0.2262±0.0111] 5 Phage display (common panning) 3 PMID:33121397 Envelope protein 2, E2 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured at 450 nm using an ELISA plate reader. ELISA values were reproduced from Figure 3 and shown. Five peptides were identified as potential binders based on their robust reactivity to the bait protein. The selected peptides appeared to interact with the crucial residues that were notably exposed on the surface of E1-E2 trimeric structure. 3651 HLNLNIYITQKH(15)[2.2067] AEAWTGFSASGV(13)[2.1833] ATLHSAHRSTHV(7)[2.2854] YPPFYMEGFLGE(1)[3.1044] SAREVMLLGDRT(1)[1.042] HPNLNIYITQKH(1)[1.4218] 6 Phage display (common panning) 4 PMID:34033860 Anti-Zearalenone (ZEN) monoclonal antibody 4D7 Zearalenone, ZEN Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance at 450 nm was determined by an automatic ELISA plate reader (Thermo, USA). If the ratio of the OD450 of test well to the OD450 of negative control well ≥2.1, the phage clone represented by this well was regarded as a positive clone recognized by the anti-ZEN antibody. Od450 was reproduced from Figure 2 and shown. After four rounds of panning, six mimotopes that could specifically bind to ZEN mAb were obtained. In order to explore the immunogenicity of these mimotopes, Balb/c mice were immunized with phages Z8 (displaying HLNLNIYITQKH), Z21 (displaying ATLHSAHRSTHV), Z35 (displaying AEAWTGFSASGV), Z8:Z21:Z35(1:1:1) and the conjugate of ZEN-bovine serum albumin (ZEN-BSA), respectively. The titers of antibodies in the mice immunized with mimotopes were 1:3200 (Z8), 1:3200 (Z21), 1:6400 (Z35), 1:6400 (1:1:1 mixture of Z8, Z21 and Z35), and the binding between serum antibodies and ZEN-OVA could be blocked by ZEN standards. These results demonstrated that the mimotopes of ZEN could induce specific antibodies against ZEN, suggesting that these displayed peptides were immunogenic. 3652 HASTGQ IAKYAT FASSSG 8 Phage display (common panning) 5 PMID:33692375 Coagulation Factor XIa (FXIa) Not determined. Not determined. Not determined. Not determined. Peptide rapidly and selectively inhibits FXIa 3653 QQGWFPG[8.7759±2.1469] QQGWVLP[7.9564±1.3562] QGQAYVL[5.1631±1.3274] GQQGWGA[4.8457±1.1427] GGQFQWV[4.055±1.1485] QGWHFGP[3.8415±0.8945] HQYGFGL[3.5587±1.3909] QWHLFAG[2.8777±0.9984] QQGWF 8 Phage display (common panning) 5 PMID:33545815 CD44/FC chimera Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Binding assay The amount of phage in the solution was determined by titer count analysis. Binding amounts are relative to that of the original phage library. The relative binding amount of bound phages was reproduced from Figure 1a and shown. These clones contained the same 4 amino acid sequence, QQGW. We concluded that this 4 amino acid sequence is important for CD44 binding. 3654 IRYDTGSYHIH 1 Phage display (common panning) 3 PMID:32449861 Anti-amyloid protein monoclonal antibody 4G8 Beta-amyloid protein 42 Not determined. Not determined. Ph.D.-12 phage display library (X12) The immune-selection of random sequences from a phage display library and sequencing to obtain the random 12 amino acids peptide library for each antibody, and then we analysed these peptides for unique and common sequences, relation to Abeta42 sequence and shape and pattern of the amino acid reaction to the antibody to predict the epitopes. Data obtained for 4G8 showed that, the sequence segment related to the putative epitope of 4G8 was LVFFAED. Nine of the ten top sequences contain the sequence RHD corresponding to the Abeta sequence from residues 5-7. Peptide 7 has the sequence IRYDTGSYHIH, which has a RYD. 3655 RVVELMDWTVLH(5)[0.7287±0.098] TMVATGLMPVLI(1)[1.5298±0.0466] SYNIIATGIHPV(1)[0.8022±0.071] KPDSCRGCLPVL(1)[0.2828±0.0392] SQDLASGLEPYF(1)[0.3538±0.0392] GSAMTWGMLAAE(1)[0.7532±0.1274] EMAVTRWDVLAI(1)[0.2191±0.0147] MIPISVGPTTKR(1)[0.2975±0.0245] 8 Phage display (competitive panning) 3 PMID:33855488 Anti-Amb a 1 IgG Amb a 1 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance was measured at 450 nm with the microtiter plate reader (Tecan Safire, Tecan Group AG, Männedorf, Switzerland). A(450 nm) were reproduced from Figure 1A and shown. Bound phages were eluted from the target antibodies either with 0.1 M glycine-HCl (pH 2.2) for 10 min followed by immediate neutralization with 1 M Tris (pH 8.0) or competitively with Amb a 1 at the final concentration of 12 μg/ml. The peptides weakly aligned with the Amb a 1 primary sequence, thus suggesting that the epitopes are conformational. When the peptides were mapped onto the surface of Amb a 1 homology model, the EpiSearch analysis predicted the location of four potential epitopic sites on surface patches centred at residues K104, S110, H214, and W312. The peptides matching to the predicted epitopes bound selectively to the IgE from pool of ragweed-allergic patients’ sera and therefore represent mimetics of Amb a 1 IgE epitopes. 3656 CLFSQGNRC(9)[0.5621±0.0956] CIMSLVGTC(7)[0.1774±0.0245] CALTQGNRC(1)[0.2338±0.049] CLVTQGNKC(1)[0.3538±0.0882] CSQGNRNWC(1)[0.3073±0.0392] CVLGLVGPC(1)[0.2338±0.0318] 6 Phage display (competitive panning) 3 PMID:33855488 Anti-Amb a 1 IgG Amb a 1 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance was measured at 450 nm with the microtiter plate reader (Tecan Safire, Tecan Group AG, Männedorf, Switzerland). A(450 nm) were reproduced from Figure 1A and shown. Bound phages were eluted from the target antibodies either with 0.1 M glycine-HCl (pH 2.2) for 10 min followed by immediate neutralization with 1 M Tris (pH 8.0) or competitively with Amb a 1 at the final concentration of 12 μg/ml. The peptides weakly aligned with the Amb a 1 primary sequence, thus suggesting that the epitopes are conformational. When the peptides were mapped onto the surface of Amb a 1 homology model, the EpiSearch analysis predicted the location of four potential epitopic sites on surface patches centred at residues K104, S110, H214, and W312. The peptides matching to the predicted epitopes bound selectively to the IgE from pool of ragweed-allergic patients’ sera and therefore represent mimetics of Amb a 1 IgE epitopes. 3657 MRTDMVI(8)[0.2828±0.0245] QDFDDIL(1)[0.5057±0.071] MRLDMEI(1)[0.2338±0.0392] 3 Phage display (competitive panning) 3 PMID:33855488 Anti-Amb a 1 IgG Amb a 1 Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Absorbance was measured at 450 nm with the microtiter plate reader (Tecan Safire, Tecan Group AG, Männedorf, Switzerland). A(450 nm) were reproduced from Figure 1A and shown. Bound phages were eluted from the target antibodies either with 0.1 M glycine-HCl (pH 2.2) for 10 min followed by immediate neutralization with 1 M Tris (pH 8.0) or competitively with Amb a 1 at the final concentration of 12 μg/ml. The peptides weakly aligned with the Amb a 1 primary sequence, thus suggesting that the epitopes are conformational. When the peptides were mapped onto the surface of Amb a 1 homology model, the EpiSearch analysis predicted the location of four potential epitopic sites on surface patches centred at residues K104, S110, H214, and W312. The peptides matching to the predicted epitopes bound selectively to the IgE from pool of ragweed-allergic patients’ sera and therefore represent mimetics of Amb a 1 IgE epitopes. 3658 SPSELGPQWSRA[1.5535±0.0806] MPSELGKQNTNV[1.0796±0.0907] FALGPSELGRWM[1.4224±0.0504] RMGPSELGPVIG[1.9214±0.1311] NNTGPFPGPSEL[1.367±0.0403] NVFGHFYGPSEL[1.5686±0.0353] GPNELGLNRPVP[1.5938±0.0756] QPNELGRIQNSH[1.3972±0.0655] SPPNALGRFLPD[1.8811±0.0655] TGSPDWRGRFIA[1.5535±0.0655] QTPDWQGRFTHT[3.5495±0.0806] VGGPDELGNNRS[0.6058±0.0655] YPDALGRLRPGP[0.5151±0.0504] DVVPMTPVERVH[0.4596±0.0958] 14 Phage display (competitive panning) 3 PMID:30315830 Anti-Api m 1 IgG Phospholipase A2, bvPLA2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance was measured at 450 nm with a microtiter plate reader. A(450 nm) were reproduced from Figure E1A and shown. Bound phages were eluted from the target antibodies either with 0.1 mol/L glycine-HCl (pH 2.2) for 10 minutes followed by immediate neutralization with 1 mol Tris (pH 8.0) or competitively with Api m 1 at the final concentration of 12 mg/mL. 3659 VSGPSEL[1.251±0.0806] LMGPSEL[1.8559±0.1059] NTGPSEL[1.619±0.0655] LTGPSEL[1.4224±0.0504] FLGPSEL[1.2661±0.0655] GLGPSEL[1.9365±0.0907] VVGPSEL[1.5938±0.0504] GWGPSEL[1.7652±0.0353] HKGPWEL[1.1301±0.0655] YPSELGP[1.498±0.0403] YPSELGR[1.1603±0.0907] NSGPSEL[1.0645±0.0554] MWGPNEL[0.8276±0.1059] GASELGK[0.999±0.0806] QASELGN[0.4748±0.0403] 15 Phage display (competitive panning) 3 PMID:30315830 Anti-Api m 1 IgG Phospholipase A2, bvPLA2 Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Absorbance was measured at 450 nm with a microtiter plate reader. A(450 nm) were reproduced from Figure E1A and shown. Bound phages were eluted from the target antibodies either with 0.1 mol/L glycine-HCl (pH 2.2) for 10 minutes followed by immediate neutralization with 1 mol Tris (pH 8.0) or competitively with Api m 1 at the final concentration of 12 mg/mL. 3660 CWTDLGRKC[2.239±0.0655] CWDSLGRAC[1.4224±0.0756] CWDAMGRAC[1.6442±0.1059] CVPDWQGRC[2.0374±0.121] CIPNNLGRC[1.251±0.0907] CIPNPIGRC[1.4325±0.1865] CYNTIGRSC[0.9335±0.0807] CNDKSKPYC[1.4325±0.121] CLDKSKPVC[1.9214±0.1159] CLDKSKPQC[1.6694±0.1462] CVDKSKPHC[2.1432±0.1059] CVDRSKPTC[1.7248±0.0504] CKDRSKPDC[2.1987±0.1159] CPDRSKPHC[1.8811±0.0907] CYDRSKPYC[1.871±0.0756] CDHSKPHTC[0.631±0.0554] CDRSKPLHC[0.2933±0.0907] 17 Phage display (competitive panning) 3 PMID:30315830 Anti-Api m 1 IgG Phospholipase A2, bvPLA2 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance was measured at 450 nm with a microtiter plate reader. A(450 nm) were reproduced from Figure E1A and shown. Bound phages were eluted from the target antibodies either with 0.1 mol/L glycine-HCl (pH 2.2) for 10 minutes followed by immediate neutralization with 1 mol Tris (pH 8.0) or competitively with Api m 1 at the final concentration of 12 mg/mL. 3661 VVRNDLSLFFIA[0.8273] ENISYVDALLTA[0.7558] 2 Phage display (competitive panning) 3 PMID:26908079 Anti-Fel d 1 IgG Major allergen I polypeptide chain 1 and 2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured at 450 nm with a microtiter plate reader. The ratio of the ELISA signal of the test phage wells to the wild-type phage (control phage) wells was calculated. The ratio value was reproduced from Figure 1 and shown. Bound phages were eluted with either rFel d 1 or with 0.1 M glycine buffer with pH 2.2. The mimotopes were computationally mapped to the allergen surface, and their IgE reactivity was confirmed using sera from cat-allergic patients. Importantly, the mimotopes showed nobasophil activation of the corresponding cat-allergic patients, which makes them good candidates forthe development of hypoallergenic vaccine. As bacteriophage particles are becoming increasingly recognized as immunogenic carriers, we constructed bacteriophage particles displaying multiple copies of each selected mimotope on major phage coat protein. These constructed phages elicited T cell-mediated immune response, which was predominated by the type 1 T cell response. Mimotopes alone contributed to the type 1 T cell response by promoting IL-2 production. Fel d 1 mimotopes, as well as their filamentous phage immunogenic carriers, represent promising candidates in the development of hypoallergenicvaccine against cat allergy. 3662 CLDLNPYYC[1.2211] CNDYFPKLC[0.8144] CGDFWPRLC[1.2191] 3 Phage display (competitive panning) 3 PMID:26908079 Anti-Fel d 1 IgG Major allergen I polypeptide chain 1 and 2 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The absorbance was measured at 450 nm with a microtiter plate reader. The ratio of the ELISA signal of the test phage wells to the wild-type phage (control phage) wells was calculated. The ratio value was reproduced from Figure 1 and shown. Bound phages were eluted with either rFel d 1 or with 0.1 M glycine buffer with pH 2.2. The mimotopes were computationally mapped to the allergen surface, and their IgE reactivity was confirmed using sera from cat-allergic patients. Importantly, the mimotopes showed nobasophil activation of the corresponding cat-allergic patients, which makes them good candidates forthe development of hypoallergenic vaccine. As bacteriophage particles are becoming increasingly recognized as immunogenic carriers, we constructed bacteriophage particles displaying multiple copies of each selected mimotope on major phage coat protein. These constructed phages elicited T cell-mediated immune response, which was predominated by the type 1 T cell response. Mimotopes alone contributed to the type 1 T cell response by promoting IL-2 production. Fel d 1 mimotopes, as well as their filamentous phage immunogenic carriers, represent promising candidates in the development of hypoallergenicvaccine against cat allergy. 3663 DHPRFNDSYNSP[1.2744±0.1438] DHPRFNRDNDVA[1.3154±0.0913] DHPRFNYVSQPW[1.1443±0.089] DHPR 3 Phage display (competitive panning) 3 PMID:37038893 Anti-Ara h 2 IgG Conglutin-7 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured at 450 nm with a microtiter plate reader. The value of OD450 was reproduced from Figure S1A and shown. Bound phages were eluted either specifically with Ara h 2 or non-specifically with 0.1 M glycine buffer with pH 2.2. We identified and selected three linear peptides (DHPRFNRDNDVA, DHPRYGP and DHPRFST), and immunoblot analyses revealed binding to lgE from peanut-allergic individuals. The peptide sequences were aligned to the disordered region corresponding to the loop between helices 2 and 3 of Ara h 2, and conformational mapping showed that the peptides match the surface of Ara h 2 and h 6 but not other peanut allergens. In ImmunoCAP inhibition experiments, the peptides significantly inhibit the binding of IgE to Ara h 2 (p < .001). In basophil and mast cell activation tests, the peptides significantly suppressed Ara h 2-induced effector cell activation (p < .05) and increased the half-maximal Ara h 2 effective concentration (p < .05). Binding of the peptides to specific IgEs did not induce activation of basophils or mast cells. 3664 DHPRFAP[1.1192±0.1438] DHPRYGP[1.1078±0.2145] DHPRFST[1.085±0.0936] DHPRFAE[1.2698±0.1825] DHPRFPL[0.875±0.0479] DHPRFSF[1.2561±0.1734] NHPRFNL[0.8887±0.0228] DHPR 7 Phage display (competitive panning) 3 PMID:37038893 Anti-Ara h 2 IgG Conglutin-7 Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The absorbance was measured at 450 nm with a microtiter plate reader. The value of OD450 was reproduced from Figure S1A and shown. Bound phages were eluted either specifically with Ara h 2 or non-specifically with 0.1 M glycine buffer with pH 2.2. We identified and selected three linear peptides (DHPRFNRDNDVA, DHPRYGP and DHPRFST), and immunoblot analyses revealed binding to lgE from peanut-allergic individuals. The peptide sequences were aligned to the disordered region corresponding to the loop between helices 2 and 3 of Ara h 2, and conformational mapping showed that the peptides match the surface of Ara h 2 and h 6 but not other peanut allergens. In ImmunoCAP inhibition experiments, the peptides significantly inhibit the binding of IgE to Ara h 2 (p < .001). In basophil and mast cell activation tests, the peptides significantly suppressed Ara h 2-induced effector cell activation (p < .05) and increased the half-maximal Ara h 2 effective concentration (p < .05). Binding of the peptides to specific IgEs did not induce activation of basophils or mast cells. 3665 ETNTAGHTSLES(1) FSPHNLTYNMDA(1) GVTDPFFDQHAE(1) LSPLSPPMRPLK(1) LVAPLDSTAPVL(1) WVNNSLATPYMS(1) YAPHLSTMLQYH(1) YDTPNNYFINYY(1) YSSPLMNDAKFP(1) 9 Phage display (common panning) 2 PMID:36746976 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3666 DTYSHQMKIRVP(1) HHGLYRMPVTIE(1) HLSYDRSVLLPT(1) LPPHAARTPSEF(1) MHPSTSWLDSTP(1) STVGPMSTLNRS(1) TSSAQLRHGPLL(1) WPDLVHTSDSRT(1) YPVRAVPNQSGQ(1) 9 Phage display (common panning) 3 PMID:36746976 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3667 AIPIWTISEVSL(1) ESWQPVHGLIPL(1) FEDSDAFRKFTM(1) FHSRMLPGRLVP(1) FNSISDAGTGCI(1) SNPFALPISTQD(1) TSLHGDPFHRMH(1) WSTERYSATRYI(1) YLDPVPKANIWL(1) 9 Phage display (common panning) 2 PMID:36746976 Vitamin D binding protein (VDBP) complex, VDBP-Complex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3668 ATFPPINSRTPA(1) FETTYMYIKSNP(1) FKTPDDSLWPHA(1) FPLSLGSVSPLN(1) GIYPFAQSSTYP(1) TNPLDARFHEPT(1) TNQSSQHVLIKE(1) TSLPFPLASRHA(1) YPDPLIESPKLG(1) 9 Phage display (common panning) 3 PMID:36746976 Vitamin D binding protein (VDBP) complex, VDBP-Complex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3669 APLPSDRSMNPS(1) ATFNSQFFSKKG(1) DPHWASLLDSVS(1) FHEIHTMPLRYA(1) HHSLIPPSPVAW(1) TNYIYRYSVDNQ(1) VLAKQHSSVPLQ(1) WPNAAPSGADSP(1) WTPDCTLSWISS(1) 9 Phage display (common panning) 2 PMID:36746976 Vitamin D binding protein, VDBP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3670 AHASDRPSQHRV(1) AQPLSVYEMDPK(1) DDIRPQLSYHGR(1) HLTATELANSYH(1) HNSGILRTMGAY(1) TSGTIFYGNSDV(1) TTSRVPDNIRLT(1) TYTLMNPSAMPQ(1) YPSSVHVQWKLL(1) 9 Phage display (common panning) 3 PMID:36746976 Vitamin D binding protein, VDBP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3671 ASSAYLKSMDPA(4) SGVYKVAYDWQH(4) ATDFLPYYHGLL(1) DSQFNKYSIATV(1) GDGNSVLKPGNW(1) GLHTSATNLYLH(1) GSAPLLTVDTSK(1) SGALHKSWYAGP(1) 8 Phage display (subtractive panning) 3 PMID:36746976 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. One point five mL of phage pool diluted with TBST (TBS + 0.1%.v/v Tween-20) to 1.0e9 Plaque Forming Units per mL (pfu/mL) from the phage library was attached to the petri dish (polystyrene). Then, the solution was rocked at room temperature (RT) for 45 min to remove phages attached to the petri dish (polystyrene) first. 3672 SGVYKVAYDWQH(7) GLHTSATNLYLH(3) TGAPPRLDARPA(1) ASSAYLKSMDPA(1) HTAHVQADRPTQ(1) 5 Phage display (subtractive panning) 4 PMID:36746976 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. One point five mL of phage pool diluted with TBST (TBS + 0.1%.v/v Tween-20) to 1.0e9 Plaque Forming Units per mL (pfu/mL) from the phage library was attached to the petri dish (polystyrene). Then, the solution was rocked at room temperature (RT) for 45 min to remove phages attached to the petri dish (polystyrene) first. 3673 SGVYKVAYDWQH(9) GLHTSATNLYLH(3) AFHPR*METQMY(1) GLHTSIPFVVPFYCH(1) GSAPLLTVDTSK(1) QWNWPVRSVANV(1) SLDGSGAALRTS(1) SNVPQVPVMGHY(1) SPFPGVMVHKNN(1) 9 Phage display (subtractive panning) 3 PMID:36746976 Vitamin D binding protein (VDBP) complex, VDBP-Complex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. One point five mL of phage pool diluted with TBST (TBS + 0.1%.v/v Tween-20) to 1.0e9 Plaque Forming Units per mL (pfu/mL) from the phage library was attached to the petri dish (polystyrene). Then, the solution was rocked at room temperature (RT) for 45 min to remove phages attached to the petri dish (polystyrene) first. The BSA-coated plate was washed 5 times with TBST. Then, the supernatant of the phage pool reacted with a petri dish (polystyrene) and was recovered and put into the BSA-coated plate. Also, it was bound by rocking at room temperature (RT) for 45 min. After that, unbound phages were collected and used for main biopanning. 3674 SGVYKVAYDWQH(7) GLHTSATNLYLH(4) GLHTPIPFVVPFYCH(1) SGVYTIPLVVPFYSH(1) SLDGAGAALRTS(1) T*TVSTENSKWW(1) 6 Phage display (subtractive panning) 4 PMID:36746976 Vitamin D binding protein (VDBP) complex, VDBP-Complex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. One point five mL of phage pool diluted with TBST (TBS + 0.1%.v/v Tween-20) to 1.0e9 Plaque Forming Units per mL (pfu/mL) from the phage library was attached to the petri dish (polystyrene). Then, the solution was rocked at room temperature (RT) for 45 min to remove phages attached to the petri dish (polystyrene) first. The BSA-coated plate was washed 5 times with TBST. Then, the supernatant of the phage pool reacted with a petri dish (polystyrene) and was recovered and put into the BSA-coated plate. Also, it was bound by rocking at room temperature (RT) for 45 min. After that, unbound phages were collected and used for main biopanning. 3675 GLHTSATNLYLH(6) SGVYKVAYDWQH(4) DSQFNKYSIATV(3) GQSEHHMRVASF(2) ASSAYLKSMDPA(1) GIATMPPTFSKQ(1) RTPEMTSLMAWG(1) VVSPDMNLLLTN(1) VVSRLPYDRVEA(1) 9 Phage display (subtractive panning) 3 PMID:36746976 Vitamin D binding protein, VDBP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. One point five mL of phage pool diluted with TBST (TBS + 0.1%.v/v Tween-20) to 1.0e9 Plaque Forming Units per mL (pfu/mL) from the phage library was attached to the petri dish (polystyrene). Then, the solution was rocked at room temperature (RT) for 45 min to remove phages attached to the petri dish (polystyrene) first. The BSA-coated plate was washed 5 times with TBST. Then, the supernatant of the phage pool reacted with a petri dish (polystyrene) and was recovered and put into the BSA-coated plate. Also, it was bound by rocking at room temperature (RT) for 45 min. After that, unbound phages were collected and used for main biopanning. 3676 GLHTSATNLYLH(2) SLDGAGAALRTS(2) DRWVARDPASIF(1) GDGNSVLKPGNW(1) HTPMSSRLSTAS(1) SGVYKVAYDWQH(1) SKGDSLPFPFAT(1) SNSIDKVNRPIN(1) VVSPDMNLLLTN(1) 9 Phage display (subtractive panning) 4 PMID:36746976 Vitamin D binding protein, VDBP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. One point five mL of phage pool diluted with TBST (TBS + 0.1%.v/v Tween-20) to 1.0e9 Plaque Forming Units per mL (pfu/mL) from the phage library was attached to the petri dish (polystyrene). Then, the solution was rocked at room temperature (RT) for 45 min to remove phages attached to the petri dish (polystyrene) first. The BSA-coated plate was washed 5 times with TBST. Then, the supernatant of the phage pool reacted with a petri dish (polystyrene) and was recovered and put into the BSA-coated plate. Also, it was bound by rocking at room temperature (RT) for 45 min. After that, unbound phages were collected and used for main biopanning. 3677 YEFHPMGNPLHR(21) SYPSNALSLHKY(1) GTGGVHPATKLT(1) HDPRMEHSLPKS(1) ISAKPIPISMRN(1) SYPSNALSLHKY(1) TASSINLHAAHE(1) TLNVPPAKRSLS(1) TSLSTAHPMLYQ(1) 9 Phage display (subtractive panning) 6 PMID:36746976 Petri dish (polystyrene, PS) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. After positive biopanning against the target, all 1.15 mL of neutralized phages were treated in a petri dish (polystyrene), and the solution was rocked at room temperature (RT) for 30 min. Only the supernatant from which the phage strongly bound to polystyrene was removed was collected. 3678 YEFHPMGNPLHR(21) HDPRMEHSLPKS(5) NTTYPTVYADKS(1) SYWYEASSYTGV(1) TNENLMVRLTHA(1) 5 Phage display (subtractive panning) 7 PMID:36746976 Petri dish (polystyrene, PS) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. After positive biopanning against the target, all 1.15 mL of neutralized phages were treated in a petri dish (polystyrene), and the solution was rocked at room temperature (RT) for 30 min. Only the supernatant from which the phage strongly bound to polystyrene was removed was collected. 3679 SLFTKQYDYFDT(5)[0.6461±0.0288] SFTKTSTFTWRD(2)[0.7244±0.0426] AMPPTDLFLHSK(2)[0.3140±0.0110] ANGTAHSTPLLW(2)[0.2769±0.0124] DRPAHGILEASL(2)[NT] FSPQNHKPNPVT(1)[0.3071±0.0193] IPQRYAPVSNLP(1)[NT] NNSHYYRNIFYT(1)[NT] TSTLYTRAQLWN(1)[NT] 9 Phage display (subtractive panning) 6 PMID:36746976 Vitamin D binding protein (VDBP) complex, VDBP-Complex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. After positive biopanning against the target, all 1.15 mL of neutralized phages were treated in a petri dish (polystyrene), and the solution was rocked at room temperature (RT) for 30 min. Only the supernatant from which the phage strongly bound to polystyrene was removed was collected. 3680 SLFTKQYDYFDT(6)[0.6461±0.0288] VPTTSHRVAVLS(3)[0.3468±0.0331] FSPQNHKPNPVT(2)[0.3071±0.0193] AMPPTDLELHSK(1)[0.3140±0.0110] IPQRYAPVSNLP(1)[NT] NNSHYYRNIFYT(1)[NT] SSAPSMVMSTLF(1)[NT] SWNHAGQPLTVV(1)[NT] SYPSNALSLHKY(1)[0.3523±0.0207] 9 Phage display (subtractive panning) 7 PMID:36746976 Vitamin D binding protein (VDBP) complex, VDBP-Complex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. After positive biopanning against the target, all 1.15 mL of neutralized phages were treated in a petri dish (polystyrene), and the solution was rocked at room temperature (RT) for 30 min. Only the supernatant from which the phage strongly bound to polystyrene was removed was collected. 3681 TGSAKFLQRDTH(3)[0.3523±0.0097] AFADGYSARRNL(1)[NT] AFKPTSGLAKLS(1)[NT] AWRPFPSATSGP(1)[NT] FAPYNNLSDNYP(1)[NT] YHGQISANAHGW(1)[NT] YSSIAPSISNAL(1)[NT] YSSTSYRALTLG(1)[NT] YTSLPTEATDRT(1)[NT] 9 Phage display (subtractive panning) 6 PMID:36746976 Vitamin D binding protein, VDBP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. After positive biopanning against the target, all 1.15 mL of neutralized phages were treated in a petri dish (polystyrene), and the solution was rocked at room temperature (RT) for 30 min. Only the supernatant from which the phage strongly bound to polystyrene was removed was collected. 3682 TGSAKFLQRDTH(3)[0.3523±0.0097] ATWWQPDARGTP(1)[NT] ATYQNWTLPHRV(1)[NT] AWRPSSASTLWN(1)[NT] GSAARTISPSLL(1)[0.4401±0.0715] SPAKPHSFYTGS(1)[NT] SVPLNSWSIFPR(1)[NT] TADVFSSSRYTR(1)[NT] VQFTPRSYQPIY(1)[NT] 9 Phage display (subtractive panning) 7 PMID:36746976 Vitamin D binding protein, VDBP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. After positive biopanning against the target, all 1.15 mL of neutralized phages were treated in a petri dish (polystyrene), and the solution was rocked at room temperature (RT) for 30 min. Only the supernatant from which the phage strongly bound to polystyrene was removed was collected.