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.