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.