3651 HLNLNIYITQKH(15)[2.2067] AEAWTGFSASGV(13)[2.1833] ATLHSAHRSTHV(7)[2.2854] YPPFYMEGFLGE(1)[3.1044] SAREVMLLGDRT(1)[1.042] HPNLNIYITQKH(1)[1.4218] 6 Phage display (common panning) 4 PMID:34033860 Anti-Zearalenone (ZEN) monoclonal antibody 4D7 Zearalenone, ZEN Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance at 450 nm was determined by an automatic ELISA plate reader (Thermo, USA). If the ratio of the OD450 of test well to the OD450 of negative control well ≥2.1, the phage clone represented by this well was regarded as a positive clone recognized by the anti-ZEN antibody. Od450 was reproduced from Figure 2 and shown. After four rounds of panning, six mimotopes that could specifically bind to ZEN mAb were obtained. In order to explore the immunogenicity of these mimotopes, Balb/c mice were immunized with phages Z8 (displaying HLNLNIYITQKH), Z21 (displaying ATLHSAHRSTHV), Z35 (displaying AEAWTGFSASGV), Z8:Z21:Z35(1:1:1) and the conjugate of ZEN-bovine serum albumin (ZEN-BSA), respectively. The titers of antibodies in the mice immunized with mimotopes were 1:3200 (Z8), 1:3200 (Z21), 1:6400 (Z35), 1:6400 (1:1:1 mixture of Z8, Z21 and Z35), and the binding between serum antibodies and ZEN-OVA could be blocked by ZEN standards. These results demonstrated that the mimotopes of ZEN could induce specific antibodies against ZEN, suggesting that these displayed peptides were immunogenic. 3652 HASTGQ IAKYAT FASSSG 8 Phage display (common panning) 5 PMID:33692375 Coagulation Factor XIa (FXIa) Not determined. Not determined. Not determined. Not determined. Peptide rapidly and selectively inhibits FXIa 3653 QQGWFPG[8.7759±2.1469] QQGWVLP[7.9564±1.3562] QGQAYVL[5.1631±1.3274] GQQGWGA[4.8457±1.1427] GGQFQWV[4.055±1.1485] QGWHFGP[3.8415±0.8945] HQYGFGL[3.5587±1.3909] QWHLFAG[2.8777±0.9984] QQGWF 8 Phage display (common panning) 5 PMID:33545815 CD44/FC chimera Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Binding assay The amount of phage in the solution was determined by titer count analysis. Binding amounts are relative to that of the original phage library. The relative binding amount of bound phages was reproduced from Figure 1a and shown. These clones contained the same 4 amino acid sequence, QQGW. We concluded that this 4 amino acid sequence is important for CD44 binding. 3654 IRYDTGSYHIH 1 Phage display (common panning) 3 PMID:32449861 Anti-amyloid protein monoclonal antibody 4G8 Beta-amyloid protein 42 Not determined. Not determined. Ph.D.-12 phage display library (X12) The immune-selection of random sequences from a phage display library and sequencing to obtain the random 12 amino acids peptide library for each antibody, and then we analysed these peptides for unique and common sequences, relation to Abeta42 sequence and shape and pattern of the amino acid reaction to the antibody to predict the epitopes. Data obtained for 4G8 showed that, the sequence segment related to the putative epitope of 4G8 was LVFFAED. Nine of the ten top sequences contain the sequence RHD corresponding to the Abeta sequence from residues 5-7. Peptide 7 has the sequence IRYDTGSYHIH, which has a RYD. 3655 RVVELMDWTVLH(5)[0.7287±0.098] TMVATGLMPVLI(1)[1.5298±0.0466] SYNIIATGIHPV(1)[0.8022±0.071] KPDSCRGCLPVL(1)[0.2828±0.0392] SQDLASGLEPYF(1)[0.3538±0.0392] GSAMTWGMLAAE(1)[0.7532±0.1274] EMAVTRWDVLAI(1)[0.2191±0.0147] MIPISVGPTTKR(1)[0.2975±0.0245] 8 Phage display (competitive panning) 3 PMID:33855488 Anti-Amb a 1 IgG Amb a 1 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance was measured at 450 nm with the microtiter plate reader (Tecan Safire, Tecan Group AG, Männedorf, Switzerland). A(450 nm) were reproduced from Figure 1A and shown. Bound phages were eluted from the target antibodies either with 0.1 M glycine-HCl (pH 2.2) for 10 min followed by immediate neutralization with 1 M Tris (pH 8.0) or competitively with Amb a 1 at the final concentration of 12 μg/ml. The peptides weakly aligned with the Amb a 1 primary sequence, thus suggesting that the epitopes are conformational. When the peptides were mapped onto the surface of Amb a 1 homology model, the EpiSearch analysis predicted the location of four potential epitopic sites on surface patches centred at residues K104, S110, H214, and W312. The peptides matching to the predicted epitopes bound selectively to the IgE from pool of ragweed-allergic patients’ sera and therefore represent mimetics of Amb a 1 IgE epitopes. 3656 CLFSQGNRC(9)[0.5621±0.0956] CIMSLVGTC(7)[0.1774±0.0245] CALTQGNRC(1)[0.2338±0.049] CLVTQGNKC(1)[0.3538±0.0882] CSQGNRNWC(1)[0.3073±0.0392] CVLGLVGPC(1)[0.2338±0.0318] 6 Phage display (competitive panning) 3 PMID:33855488 Anti-Amb a 1 IgG Amb a 1 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance was measured at 450 nm with the microtiter plate reader (Tecan Safire, Tecan Group AG, Männedorf, Switzerland). A(450 nm) were reproduced from Figure 1A and shown. Bound phages were eluted from the target antibodies either with 0.1 M glycine-HCl (pH 2.2) for 10 min followed by immediate neutralization with 1 M Tris (pH 8.0) or competitively with Amb a 1 at the final concentration of 12 μg/ml. The peptides weakly aligned with the Amb a 1 primary sequence, thus suggesting that the epitopes are conformational. When the peptides were mapped onto the surface of Amb a 1 homology model, the EpiSearch analysis predicted the location of four potential epitopic sites on surface patches centred at residues K104, S110, H214, and W312. The peptides matching to the predicted epitopes bound selectively to the IgE from pool of ragweed-allergic patients’ sera and therefore represent mimetics of Amb a 1 IgE epitopes. 3657 MRTDMVI(8)[0.2828±0.0245] QDFDDIL(1)[0.5057±0.071] MRLDMEI(1)[0.2338±0.0392] 3 Phage display (competitive panning) 3 PMID:33855488 Anti-Amb a 1 IgG Amb a 1 Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Absorbance was measured at 450 nm with the microtiter plate reader (Tecan Safire, Tecan Group AG, Männedorf, Switzerland). A(450 nm) were reproduced from Figure 1A and shown. Bound phages were eluted from the target antibodies either with 0.1 M glycine-HCl (pH 2.2) for 10 min followed by immediate neutralization with 1 M Tris (pH 8.0) or competitively with Amb a 1 at the final concentration of 12 μg/ml. The peptides weakly aligned with the Amb a 1 primary sequence, thus suggesting that the epitopes are conformational. When the peptides were mapped onto the surface of Amb a 1 homology model, the EpiSearch analysis predicted the location of four potential epitopic sites on surface patches centred at residues K104, S110, H214, and W312. The peptides matching to the predicted epitopes bound selectively to the IgE from pool of ragweed-allergic patients’ sera and therefore represent mimetics of Amb a 1 IgE epitopes. 3658 SPSELGPQWSRA[1.5535±0.0806] MPSELGKQNTNV[1.0796±0.0907] FALGPSELGRWM[1.4224±0.0504] RMGPSELGPVIG[1.9214±0.1311] NNTGPFPGPSEL[1.367±0.0403] NVFGHFYGPSEL[1.5686±0.0353] GPNELGLNRPVP[1.5938±0.0756] QPNELGRIQNSH[1.3972±0.0655] SPPNALGRFLPD[1.8811±0.0655] TGSPDWRGRFIA[1.5535±0.0655] QTPDWQGRFTHT[3.5495±0.0806] VGGPDELGNNRS[0.6058±0.0655] YPDALGRLRPGP[0.5151±0.0504] DVVPMTPVERVH[0.4596±0.0958] 14 Phage display (competitive panning) 3 PMID:30315830 Anti-Api m 1 IgG Phospholipase A2, bvPLA2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbance was measured at 450 nm with a microtiter plate reader. A(450 nm) were reproduced from Figure E1A and shown. Bound phages were eluted from the target antibodies either with 0.1 mol/L glycine-HCl (pH 2.2) for 10 minutes followed by immediate neutralization with 1 mol Tris (pH 8.0) or competitively with Api m 1 at the final concentration of 12 mg/mL. 3659 VSGPSEL[1.251±0.0806] LMGPSEL[1.8559±0.1059] NTGPSEL[1.619±0.0655] LTGPSEL[1.4224±0.0504] FLGPSEL[1.2661±0.0655] GLGPSEL[1.9365±0.0907] VVGPSEL[1.5938±0.0504] GWGPSEL[1.7652±0.0353] HKGPWEL[1.1301±0.0655] YPSELGP[1.498±0.0403] YPSELGR[1.1603±0.0907] NSGPSEL[1.0645±0.0554] MWGPNEL[0.8276±0.1059] GASELGK[0.999±0.0806] QASELGN[0.4748±0.0403] 15 Phage display (competitive panning) 3 PMID:30315830 Anti-Api m 1 IgG Phospholipase A2, bvPLA2 Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Absorbance was measured at 450 nm with a microtiter plate reader. A(450 nm) were reproduced from Figure E1A and shown. Bound phages were eluted from the target antibodies either with 0.1 mol/L glycine-HCl (pH 2.2) for 10 minutes followed by immediate neutralization with 1 mol Tris (pH 8.0) or competitively with Api m 1 at the final concentration of 12 mg/mL. 3660 CWTDLGRKC[2.239±0.0655] CWDSLGRAC[1.4224±0.0756] CWDAMGRAC[1.6442±0.1059] CVPDWQGRC[2.0374±0.121] CIPNNLGRC[1.251±0.0907] CIPNPIGRC[1.4325±0.1865] CYNTIGRSC[0.9335±0.0807] CNDKSKPYC[1.4325±0.121] CLDKSKPVC[1.9214±0.1159] CLDKSKPQC[1.6694±0.1462] CVDKSKPHC[2.1432±0.1059] CVDRSKPTC[1.7248±0.0504] CKDRSKPDC[2.1987±0.1159] CPDRSKPHC[1.8811±0.0907] CYDRSKPYC[1.871±0.0756] CDHSKPHTC[0.631±0.0554] CDRSKPLHC[0.2933±0.0907] 17 Phage display (competitive panning) 3 PMID:30315830 Anti-Api m 1 IgG Phospholipase A2, bvPLA2 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Absorbance was measured at 450 nm with a microtiter plate reader. A(450 nm) were reproduced from Figure E1A and shown. Bound phages were eluted from the target antibodies either with 0.1 mol/L glycine-HCl (pH 2.2) for 10 minutes followed by immediate neutralization with 1 mol Tris (pH 8.0) or competitively with Api m 1 at the final concentration of 12 mg/mL. 3661 VVRNDLSLFFIA[0.8273] ENISYVDALLTA[0.7558] 2 Phage display (competitive panning) 3 PMID:26908079 Anti-Fel d 1 IgG Major allergen I polypeptide chain 1 and 2 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured at 450 nm with a microtiter plate reader. The ratio of the ELISA signal of the test phage wells to the wild-type phage (control phage) wells was calculated. The ratio value was reproduced from Figure 1 and shown. Bound phages were eluted with either rFel d 1 or with 0.1 M glycine buffer with pH 2.2. The mimotopes were computationally mapped to the allergen surface, and their IgE reactivity was confirmed using sera from cat-allergic patients. Importantly, the mimotopes showed nobasophil activation of the corresponding cat-allergic patients, which makes them good candidates forthe development of hypoallergenic vaccine. As bacteriophage particles are becoming increasingly recognized as immunogenic carriers, we constructed bacteriophage particles displaying multiple copies of each selected mimotope on major phage coat protein. These constructed phages elicited T cell-mediated immune response, which was predominated by the type 1 T cell response. Mimotopes alone contributed to the type 1 T cell response by promoting IL-2 production. Fel d 1 mimotopes, as well as their filamentous phage immunogenic carriers, represent promising candidates in the development of hypoallergenicvaccine against cat allergy. 3662 CLDLNPYYC[1.2211] CNDYFPKLC[0.8144] CGDFWPRLC[1.2191] 3 Phage display (competitive panning) 3 PMID:26908079 Anti-Fel d 1 IgG Major allergen I polypeptide chain 1 and 2 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The absorbance was measured at 450 nm with a microtiter plate reader. The ratio of the ELISA signal of the test phage wells to the wild-type phage (control phage) wells was calculated. The ratio value was reproduced from Figure 1 and shown. Bound phages were eluted with either rFel d 1 or with 0.1 M glycine buffer with pH 2.2. The mimotopes were computationally mapped to the allergen surface, and their IgE reactivity was confirmed using sera from cat-allergic patients. Importantly, the mimotopes showed nobasophil activation of the corresponding cat-allergic patients, which makes them good candidates forthe development of hypoallergenic vaccine. As bacteriophage particles are becoming increasingly recognized as immunogenic carriers, we constructed bacteriophage particles displaying multiple copies of each selected mimotope on major phage coat protein. These constructed phages elicited T cell-mediated immune response, which was predominated by the type 1 T cell response. Mimotopes alone contributed to the type 1 T cell response by promoting IL-2 production. Fel d 1 mimotopes, as well as their filamentous phage immunogenic carriers, represent promising candidates in the development of hypoallergenicvaccine against cat allergy. 3663 DHPRFNDSYNSP[1.2744±0.1438] DHPRFNRDNDVA[1.3154±0.0913] DHPRFNYVSQPW[1.1443±0.089] DHPR 3 Phage display (competitive panning) 3 PMID:37038893 Anti-Ara h 2 IgG Conglutin-7 Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured at 450 nm with a microtiter plate reader. The value of OD450 was reproduced from Figure S1A and shown. Bound phages were eluted either specifically with Ara h 2 or non-specifically with 0.1 M glycine buffer with pH 2.2. We identified and selected three linear peptides (DHPRFNRDNDVA, DHPRYGP and DHPRFST), and immunoblot analyses revealed binding to lgE from peanut-allergic individuals. The peptide sequences were aligned to the disordered region corresponding to the loop between helices 2 and 3 of Ara h 2, and conformational mapping showed that the peptides match the surface of Ara h 2 and h 6 but not other peanut allergens. In ImmunoCAP inhibition experiments, the peptides significantly inhibit the binding of IgE to Ara h 2 (p < .001). In basophil and mast cell activation tests, the peptides significantly suppressed Ara h 2-induced effector cell activation (p < .05) and increased the half-maximal Ara h 2 effective concentration (p < .05). Binding of the peptides to specific IgEs did not induce activation of basophils or mast cells. 3664 DHPRFAP[1.1192±0.1438] DHPRYGP[1.1078±0.2145] DHPRFST[1.085±0.0936] DHPRFAE[1.2698±0.1825] DHPRFPL[0.875±0.0479] DHPRFSF[1.2561±0.1734] NHPRFNL[0.8887±0.0228] DHPR 7 Phage display (competitive panning) 3 PMID:37038893 Anti-Ara h 2 IgG Conglutin-7 Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The absorbance was measured at 450 nm with a microtiter plate reader. The value of OD450 was reproduced from Figure S1A and shown. Bound phages were eluted either specifically with Ara h 2 or non-specifically with 0.1 M glycine buffer with pH 2.2. We identified and selected three linear peptides (DHPRFNRDNDVA, DHPRYGP and DHPRFST), and immunoblot analyses revealed binding to lgE from peanut-allergic individuals. The peptide sequences were aligned to the disordered region corresponding to the loop between helices 2 and 3 of Ara h 2, and conformational mapping showed that the peptides match the surface of Ara h 2 and h 6 but not other peanut allergens. In ImmunoCAP inhibition experiments, the peptides significantly inhibit the binding of IgE to Ara h 2 (p < .001). In basophil and mast cell activation tests, the peptides significantly suppressed Ara h 2-induced effector cell activation (p < .05) and increased the half-maximal Ara h 2 effective concentration (p < .05). Binding of the peptides to specific IgEs did not induce activation of basophils or mast cells. 3665 ETNTAGHTSLES(1) FSPHNLTYNMDA(1) GVTDPFFDQHAE(1) LSPLSPPMRPLK(1) LVAPLDSTAPVL(1) WVNNSLATPYMS(1) YAPHLSTMLQYH(1) YDTPNNYFINYY(1) YSSPLMNDAKFP(1) 9 Phage display (common panning) 2 PMID:36746976 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3666 DTYSHQMKIRVP(1) HHGLYRMPVTIE(1) HLSYDRSVLLPT(1) LPPHAARTPSEF(1) MHPSTSWLDSTP(1) STVGPMSTLNRS(1) TSSAQLRHGPLL(1) WPDLVHTSDSRT(1) YPVRAVPNQSGQ(1) 9 Phage display (common panning) 3 PMID:36746976 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3667 AIPIWTISEVSL(1) ESWQPVHGLIPL(1) FEDSDAFRKFTM(1) FHSRMLPGRLVP(1) FNSISDAGTGCI(1) SNPFALPISTQD(1) TSLHGDPFHRMH(1) WSTERYSATRYI(1) YLDPVPKANIWL(1) 9 Phage display (common panning) 2 PMID:36746976 Vitamin D binding protein (VDBP) complex, VDBP-Complex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3668 ATFPPINSRTPA(1) FETTYMYIKSNP(1) FKTPDDSLWPHA(1) FPLSLGSVSPLN(1) GIYPFAQSSTYP(1) TNPLDARFHEPT(1) TNQSSQHVLIKE(1) TSLPFPLASRHA(1) YPDPLIESPKLG(1) 9 Phage display (common panning) 3 PMID:36746976 Vitamin D binding protein (VDBP) complex, VDBP-Complex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3669 APLPSDRSMNPS(1) ATFNSQFFSKKG(1) DPHWASLLDSVS(1) FHEIHTMPLRYA(1) HHSLIPPSPVAW(1) TNYIYRYSVDNQ(1) VLAKQHSSVPLQ(1) WPNAAPSGADSP(1) WTPDCTLSWISS(1) 9 Phage display (common panning) 2 PMID:36746976 Vitamin D binding protein, VDBP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3670 AHASDRPSQHRV(1) AQPLSVYEMDPK(1) DDIRPQLSYHGR(1) HLTATELANSYH(1) HNSGILRTMGAY(1) TSGTIFYGNSDV(1) TTSRVPDNIRLT(1) TYTLMNPSAMPQ(1) YPSSVHVQWKLL(1) 9 Phage display (common panning) 3 PMID:36746976 Vitamin D binding protein, VDBP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. 3671 ASSAYLKSMDPA(4) SGVYKVAYDWQH(4) ATDFLPYYHGLL(1) DSQFNKYSIATV(1) GDGNSVLKPGNW(1) GLHTSATNLYLH(1) GSAPLLTVDTSK(1) SGALHKSWYAGP(1) 8 Phage display (subtractive panning) 3 PMID:36746976 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. One point five mL of phage pool diluted with TBST (TBS + 0.1%.v/v Tween-20) to 1.0e9 Plaque Forming Units per mL (pfu/mL) from the phage library was attached to the petri dish (polystyrene). Then, the solution was rocked at room temperature (RT) for 45 min to remove phages attached to the petri dish (polystyrene) first. 3672 SGVYKVAYDWQH(7) GLHTSATNLYLH(3) TGAPPRLDARPA(1) ASSAYLKSMDPA(1) HTAHVQADRPTQ(1) 5 Phage display (subtractive panning) 4 PMID:36746976 Bovine serum albumin (BSA) Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. One point five mL of phage pool diluted with TBST (TBS + 0.1%.v/v Tween-20) to 1.0e9 Plaque Forming Units per mL (pfu/mL) from the phage library was attached to the petri dish (polystyrene). Then, the solution was rocked at room temperature (RT) for 45 min to remove phages attached to the petri dish (polystyrene) first. 3673 SGVYKVAYDWQH(9) GLHTSATNLYLH(3) AFHPR*METQMY(1) GLHTSIPFVVPFYCH(1) GSAPLLTVDTSK(1) QWNWPVRSVANV(1) SLDGSGAALRTS(1) SNVPQVPVMGHY(1) SPFPGVMVHKNN(1) 9 Phage display (subtractive panning) 3 PMID:36746976 Vitamin D binding protein (VDBP) complex, VDBP-Complex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. One point five mL of phage pool diluted with TBST (TBS + 0.1%.v/v Tween-20) to 1.0e9 Plaque Forming Units per mL (pfu/mL) from the phage library was attached to the petri dish (polystyrene). Then, the solution was rocked at room temperature (RT) for 45 min to remove phages attached to the petri dish (polystyrene) first. The BSA-coated plate was washed 5 times with TBST. Then, the supernatant of the phage pool reacted with a petri dish (polystyrene) and was recovered and put into the BSA-coated plate. Also, it was bound by rocking at room temperature (RT) for 45 min. After that, unbound phages were collected and used for main biopanning. 3674 SGVYKVAYDWQH(7) GLHTSATNLYLH(4) GLHTPIPFVVPFYCH(1) SGVYTIPLVVPFYSH(1) SLDGAGAALRTS(1) T*TVSTENSKWW(1) 6 Phage display (subtractive panning) 4 PMID:36746976 Vitamin D binding protein (VDBP) complex, VDBP-Complex Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. One point five mL of phage pool diluted with TBST (TBS + 0.1%.v/v Tween-20) to 1.0e9 Plaque Forming Units per mL (pfu/mL) from the phage library was attached to the petri dish (polystyrene). Then, the solution was rocked at room temperature (RT) for 45 min to remove phages attached to the petri dish (polystyrene) first. The BSA-coated plate was washed 5 times with TBST. Then, the supernatant of the phage pool reacted with a petri dish (polystyrene) and was recovered and put into the BSA-coated plate. Also, it was bound by rocking at room temperature (RT) for 45 min. After that, unbound phages were collected and used for main biopanning. 3675 GLHTSATNLYLH(6) SGVYKVAYDWQH(4) DSQFNKYSIATV(3) GQSEHHMRVASF(2) ASSAYLKSMDPA(1) GIATMPPTFSKQ(1) RTPEMTSLMAWG(1) VVSPDMNLLLTN(1) VVSRLPYDRVEA(1) 9 Phage display (subtractive panning) 3 PMID:36746976 Vitamin D binding protein, VDBP Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Data are expressed as the means of the absorbance value at (410 ± SD) nm. One point five mL of phage pool diluted with TBST (TBS + 0.1%.v/v Tween-20) to 1.0e9 Plaque Forming Units per mL (pfu/mL) from the phage library was attached to the petri dish (polystyrene). Then, the solution was rocked at room temperature (RT) for 45 min to remove phages attached to the petri dish (polystyrene) first. The BSA-coated plate was washed 5 times with TBST. Then, the supernatant of the phage pool reacted with a petri dish (polystyrene) and was recovered and put into the BSA-coated plate. Also, it was bound by rocking at room temperature (RT) for 45 min. After that, unbound phages were collected and used for main biopanning.