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.