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.