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.