1601 KPWMTWQWVVEKSSTEGFRT(2) GDELGWNWVTIWPKTLSSRV(3) 2 Phage display (subtractive panning) 10 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 4°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. Clearing with autologous EBV-immortalized cell line was applied in rounds 6-10. 1602 KPWMTWQWVVEKSSTEGFRT(5) 1 Phage display (subtractive panning) 10 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 4°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. Clearing with autologous EBV-immortalized cell line was applied in rounds 6-10. 1603 ASSWKSFWHPNPVGMTPASS(4) 1 Phage display (subtractive panning) 10 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 37°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. Clearing with autologous EBV-immortalized cell line was applied in rounds 6-10. 1604 LTSPPKEARPSTTVGKSGRE(1) MARTVTANVPGMGEGMVVVP(3) 2 Phage display (subtractive panning) 10 PMID:14500347 CD19+ cells from patient #25 Not determined. Not determined. Not determined. ON543 phage display library (X20) The cell were incubated for 1 h with the phage at 37°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. Clearing with autologous EBV-immortalized cell line was applied in rounds 6-10. 1605 LAVTRPSPNKYATVNKSAQA(1) RTSTGPEARDAWMWYWKPPA(3) FGWTWEDSSSNSLYDMSFPH(1) GARSPIRESFGNTVPFDTRF(1) TGQGEREHREGRYTNSEGSD(1) VQTREAGGMYGGPDWWWEWS(1) ARAFSYWSVTDTERVKFFVP(1) LQIGEGVELVMARPIDSDSG(1) 8 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 16; R2, patient 25; R3, patient 42; R4, patient 19; R5, patient 49. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 4°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1606 MGSRSAVGDFESAEGSRRP(3) MARMSTSEVPPVRIATSHSR(1) ISWDIWRWWYTSEDRDAGSA(3) EFGHGSPYETRQHSQARRWL(1) MAQKWGGGQALQGYHIGANA(1) 5 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 16; R2, patient 25; R3, patient 42; R4, patient 19; R5, patient 49. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 25°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1607 VWGMTTSDHQRKTERLDSPE(1) GTFLHSRSQTSAEERAGGLG(1) GWWQFWDWSARRQDRGEALL(4) MAMGGKPERPADSDNVQVRG(1) QTIWRGHAASVNDSTTVSRT(1) MTSAQTSEKLKAETDRHTAE(1) 6 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 16; R2, patient 25; R3, patient 42; R4, patient 19; R5, patient 49. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 37°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1608 GAGNHPDAVSYPDILVKPRL(4) TAYTTGIRWSSQLQRLQHAS(1) 2 Phage display (subtractive panning) 4 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-4. The cell were incubated for 1 h with the phage at 4°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1609 GAGNHPDAVSYPDILVKPRL(3) TAYTTGIRWSSQLQRLQHAS(1) TDAMLGATVREHLEAMALVG(1) 3 Phage display (subtractive panning) 4 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-4. The cell were incubated for 1 h with the phage at 4°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1610 GAGNHPDAVSYPDILVKPRL(2) TAYTTGIRWSSQLQRLQHAS(1) MPHLDRVLWKPSGIRDLVSE(1) 3 Phage display (subtractive panning) 4 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-4. The cell were incubated for 1 h with the phage at 37°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1611 GAGNHPDAVSYPDILVKPRL(1) TAYTTGIRWSSQLQRLQHAS(2) VTGAVWMVLEESSHVLL(1) EYRVENRIQPIRAYSGSLNR(1) 4 Phage display (subtractive panning) 4 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-4. The cell were incubated for 1 h with the phage at 37°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1612 KPWMTWQWVVEKSSTEGFRT(6) GAGNHPDAVSYPDILVKPRL(1) TQGRMAFYQTSVVLSSADST(1) 3 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 38; R5, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 4°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1613 TAYTTGIRWSSQLQRLQHAS(1) 1 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 38; R5, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 4°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1614 GAGNHPDAVSYPDILVKPRL(2) TAYTTGIRWSSQLQRLQHAS(1) TQGRMAFYQTSVVLSSADST(1) WGTKPTTQWRKPQLQEEVRP(3) 4 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 38; R5, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 37°C. Cell surface phage were recovered by two acid elutions. The acid fraction was neutralized. 1615 TAYTTGIRWSSQLQRLQHAS(2) WGTKPTTQWRKPQLQEEVRP(3) MGRTVQSGDGTPAQTQPSVN(3) 3 Phage display (subtractive panning) 5 PMID:14500347 CD19+ cells Not determined. Not determined. Not determined. ON543 phage display library (X20) Cells from different patients were used at the indicated rounds of selection (e.g., R1 = round 1). Selection target referring to the patient: R1, patient 25; R2, patient 16; R3, patient 18; R4, patient 38; R5, patient 43. Clearing with autologous EBV-immortalized cell line was applied in rounds 2-5. The cell were incubated for 1 h with the phage at 37°C. The tightly associated phage or internalized phage were recovered by hypotonic lysis of the cells in 30 mM Tris (pH 8) on ice for 30 min combined with brief vortexing. 1616 RGDKSRPPVWYVEGE(7)[1.085 ± 0.011, 37.181 ± 5.147, 1.069 ± 0.051, 0.026 ± 0.005] VQRGAWRWPVVTNE(4)[0.242 ± 0.017, 10.291 ± 1.133, 1.083 ± 0.031, 0.012] PAGRQARRPATVPEG(1)[0.074, 12.289, 1.013 ± 0.016, 0.025] PEWWLEEPPFISSGK(1)[0.199 ± 0.014, 4.433 ± 0.346, 1.083 ± 0.041, 0.005 ± 0.002] 4 Phage display (common panning) 2 PMID:20708963 Anti-CPS monoclonal antibody 13D9 Capsular polysaccharide, CPS Not determined. Not determined. X15 PC89 phage display library ELISA The affinities of selected phages to anti-CPS monoclonal antibody 13D9 were measured by phage ELISA, phage UltramicroELISA (UMELISA), phage sandwich ELISA and inhibition ELISA. In phage ELISA, the absorbance at 492 nm was determined. The absorbance of the control phage PC89 was 0.086 ± 0.011. In UMELISA, fluorescence units (FU) were read in a photometer-fluorimeter plate reader PR-521. The fluorescence units of the control phage PC89 were 2.436. In sandwich ELISA, the absorbance at 492 nm was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.087 ± 0.020 and 0.077 ± 0.020, respectively. In inhibition ELISA, the absorbance at 492 was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.119 ± 0.008 and 0.188 ± 0.006, respectively. All data were expressed as means ± SD and were reproduced from the graph. Immunization of mice with the phage-displayed peptides elicited anti-peptide IgG antibodies. 1617 RGDKSRPPVWYVEGE(20)[0.380 ± 0.033, 1.069 ± 0.051, 0.026 ± 0.005] 1 Phage display (common panning) 3 PMID:20708963 Anti-CPS monoclonal antibody 13D9 Capsular polysaccharide, CPS Not determined. Not determined. X15 PC89 phage display library ELISA The affinities of selected phages to anti-CPS monoclonal antibody 13D9 were measured by phage ELISA, phage UltramicroELISA (UMELISA), phage sandwich ELISA and inhibition ELISA. In phage ELISA, the absorbance at 492 nm was determined. The absorbance of the control phage PC89 was 0.086 ± 0.011. In UMELISA, fluorescence units (FU) were read in a photometer-fluorimeter plate reader PR-521. The fluorescence units of the control phage PC89 were 2.436. In sandwich ELISA, the absorbance at 492 nm was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.087 ± 0.020 and 0.077 ± 0.020, respectively. In inhibition ELISA, the absorbance at 492 was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.119 ± 0.008 and 0.188 ± 0.006, respectively. All data were expressed as means ± SD and were reproduced from the graph. Immunization of mice with the phage-displayed peptides elicited anti-peptide IgG antibodies. 1618 RGDKSRPPVWYVEGE(25)[0.380 ± 0.033, 1.069 ± 0.051, 0.026 ± 0.005] 1 Phage display (common panning) 4 PMID:20708963 Anti-CPS monoclonal antibody 13D9 Capsular polysaccharide, CPS Not determined. Not determined. X15 PC89 phage display library ELISA The affinities of selected phages to anti-CPS monoclonal antibody 13D9 were measured by phage ELISA, phage UltramicroELISA (UMELISA), phage sandwich ELISA and inhibition ELISA. In phage ELISA, the absorbance at 492 nm was determined. The absorbance of the control phage PC89 was 0.086 ± 0.011. In UMELISA, fluorescence units (FU) were read in a photometer-fluorimeter plate reader PR-521. The fluorescence units of the control phage PC89 were 2.436. In sandwich ELISA, the absorbance at 492 nm was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.087 ± 0.020 and 0.077 ± 0.020, respectively. In inhibition ELISA, the absorbance at 492 was determined. The absorbances of the phage 2L-18, behaving as the control phage PC89, and PC89 were 0.119 ± 0.008 and 0.188 ± 0.006, respectively. All data were expressed as means ± SD and were reproduced from the graph. Immunization of mice with the phage-displayed peptides elicited anti-peptide IgG antibodies. 1619 ATNAEPS(1) GGKRLPR(1) GRKLKPR(1) GTGKRPR(1) KVSRPVR(1) LDEEPPR(1) LGGKKKR(1) NKVRLPR(1) PSRRKLR(1) REGRVRR(1) RGGRGPR(1) RGGRPAR(1) RKGRPSR(1) RVMRGPR(1) RWEKVPR(1) TVKKNKR(1) VLTRGLR(1) VMHKPAR(1) VMVRPPR(1) VRSKAPR(1) VSERRPR(1) [RK]-x(2)-R 21 Phage display (common panning) 3 PMID:21486998 Neuropilin-1 Not determined. Not determined. Not determined. X7 T7 phage display library PPC-1 cells were selected as a model because they express high levels of NRP-1. Selection was performed against PPC-1 cells by the microfluidic phage selection (MiPS) platform. The ability of this system to directly target membrane-bound proteins on cell surfaces is significant because most of these proteins are notoriously difficult to produce using recombinant technology. Authors show that this system offers significant advantages compared to conventional biopanning methods. Authors found that the MiPS system enables more efficient enrichment of phage displaying this higher-affinity motif: 90% of the phage selected using the MiPS contained (R/K)XX(R/K) motifs, while 5% displayed lower affinity XXXR motifs. 1620 AQVRGPR(2) RTPR(1) STVRRRR(1) MKIRGPR(1) GRVRGVR(1) LKIERSPR(1) RRKR(1) GKPRAKR(1) AMKRVAR(1) QRKGRR(1) VRKFVIR(1) TNRVRRR(1) RGSQRAR(1) KRLNSGR(1) KMRSSLR(1) IGQDSHR(1) NETRSGN(1) RKRRSVL(1) EVEGCEL(1) VGKKLP(1) [RK]-x(2)-[RK] 20 Phage display (common panning) 3 PMID:21486998 Neuropilin-1 Not determined. Not determined. Not determined. X7 T7 phage display library PPC-1 cells were selected as a model because they express high levels of NRP-1. Authors found that conventional cell suspension-based panning yielded at best 52% (R/K)XX(R/K) phage and 29% XXXR phage. 1621 RIGRPLR(1)[5567, 0.78] GGKRPAR(1)[4645, 1.51] FNRKERR(1)[2650, NT] GKGRAQR(1)[1045, NT] SRAKKPR(1)[912, NT] NGARKPR(1)[817, 3.29] LRKRAAR(1)[581, NT] KKVRVVR(1)[474, NT] GKVGSRR(1)[377, NT] VYKARTR(1)[293, 10.9] LSSAQDR(1)[14, NT] VGSGKRS(1)[1.4, NT] SMTARSV(1)[0.8, NT] GGRGARA(1)[0.6, NT] RVRAGHH(1)[0.8, NT] [RK]-x(2)-R 15 Phage display (common panning) 3 PMID:21486998 Neuropilin-1 Not determined. Not determined. Not determined. X7 T7 phage display library Competition experiment Fifteen phage clones were randomly picked from the R3 pool and their binding to PPC-1 relative to M21 melanoma cells was measured. Of note, in contrast to PPC-1 cells, M21 cells express only minimal amounts of NRP-1 protein. The binding fold was shown. Besides, the equilibrium dissociation constants (Kd, μM) of the peptide sequences for binding to the NRP-1 protein were measured and shown. Three rounds of selection in MiPS device revealed peptides P7 (RIGRPLR) and P4 (GGKRPAR) exhibiting higher affinity and specificity in binding to cells expressing NRP-1 compared with the well known CR (RPARPAR) with a Kd of 2.79 μM. NT denotes not tested. PPC-1 cells were selected as a model because they express high levels of NRP-1. Selection was performed against PPC-1 cells by the microfluidic phage selection (MiPS) platform. The ability of this system to directly target membrane-bound proteins on cell surfaces is significant because most of these proteins are notoriously difficult to produce using recombinant technology. Authors show that this system offers significant advantages compared to conventional biopanning methods. 1622 AKAFSYWTPFDGKSLYSST(1)[0.581 ± 0.016] IVTARARHDWNWVQRSSRSL(1)[0.047] TYNHDYKTGMVIYSPFMTYP(1)[1.791 ± 0.010] 3 Phage display (common panning) 3 PMID:21697352 Anti-NSP2 monoclonal antibody Non-structural protein 2, NSP2 Not determined. Not determined. X20 fUSE5 phage display library ELISA In phage ELISA, the absorbance at 490 nm was determined. Data shown were reproduced from the graph and expressed as means ± SD. BLAST and ClustalW2 analysis of the phage clone TYNHDYKTGMVIYSPFMTYP peptide and NSP2 protein sequences located a motif (244)T-(Y/F)-Φ-Φ-Φ-X-K-Φ-G(252), in the C-terminal domain of the protein, where Φ is a hydrophilic residue and amino acids at position X249 did not display conserved/similar properties. This motif was found to be conserved from analyzing an alignment of Australian and GenBank human NSP2 sequences, which also suggests that antibodies against NSP2 may provide heterotypic protection. 1623 TYNHDYKTGMVIYSPFMTYP 1 Phage display (common panning) 4 PMID:21697352 Anti-NSP2 monoclonal antibody Non-structural protein 2, NSP2 Not determined. Not determined. X20 fUSE5 phage display library BLAST and ClustalW2 analysis of the phage clone TYNHDYKTGMVIYSPFMTYP peptide and NSP2 protein sequences located a motif (244)T-(Y/F)-Φ-Φ-Φ-X-K-Φ-G(252), in the C-terminal domain of the protein, where Φ is a hydrophilic residue and amino acids at position X249 did not display conserved/similar properties. This motif was found to be conserved from analyzing an alignment of Australian and GenBank human NSP2 sequences, which also suggests that antibodies against NSP2 may provide heterotypic protection. 1624 TYNHDYKTGMVIYSPFMTYP 1 Phage display (common panning) 5 PMID:21697352 Anti-NSP2 monoclonal antibody Non-structural protein 2, NSP2 Not determined. Not determined. X20 fUSE5 phage display library BLAST and ClustalW2 analysis of the phage clone TYNHDYKTGMVIYSPFMTYP peptide and NSP2 protein sequences located a motif (244)T-(Y/F)-Φ-Φ-Φ-X-K-Φ-G(252), in the C-terminal domain of the protein, where Φ is a hydrophilic residue and amino acids at position X249 did not display conserved/similar properties. This motif was found to be conserved from analyzing an alignment of Australian and GenBank human NSP2 sequences, which also suggests that antibodies against NSP2 may provide heterotypic protection. 1625 TYNHDYKTGMVIYSPFMTYP 1 Phage display (common panning) 6 PMID:21697352 Anti-NSP2 monoclonal antibody Non-structural protein 2, NSP2 Not determined. Not determined. X20 fUSE5 phage display library BLAST and ClustalW2 analysis of the phage clone TYNHDYKTGMVIYSPFMTYP peptide and NSP2 protein sequences located a motif (244)T-(Y/F)-Φ-Φ-Φ-X-K-Φ-G(252), in the C-terminal domain of the protein, where Φ is a hydrophilic residue and amino acids at position X249 did not display conserved/similar properties. This motif was found to be conserved from analyzing an alignment of Australian and GenBank human NSP2 sequences, which also suggests that antibodies against NSP2 may provide heterotypic protection.