2801 TPSYDTYAAELR(16) SVSVGMKPSPRP(4) KSIALKNTNPHA(1) SPCVQCSSGLCP(1) PCLKMGIHTTKR(1) YGLHRQAACPLT(1) 6 Phage display (in vivo) 3 PMID:25408466 Cerebrospinal fluid Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) In the first round, the rats (n = 3) were injected intravenously (iv) with e12 pfu of the library suspended in 100 μl TBS. The phage was allowed to circulate for some time in vivo. The rats were then anesthetized using 5 % chloral hydrate (0.4 g/kg), and phage-containing supernatant was recovered from the cerebrospinal fluid, amplified by infecting competent bacteria (Escherichia coli ER2738), titered and pooled for the next round of screening. Subsequent screening rounds were conducted by intravenously injecting the newly amplified phage (1 × e12 pfu in 100 μl TBS) and repeating the procedures described above. After labeling with FITC, TPSYDTYAAELR, referred to as the TPS peptide, demonstrated significantly greater brain accumulation efficiency. The TPS peptide represents a previously unreported promising motif that can be used to design a drug delivery system that can cross the blood–cerebrospinalfluid barrier (BCSFB). 2802 SWPLYSDASGLG(11)[0.72 ± 0.17] VSLHDDASGTHR(2)[0.92/0.85] WLGESMISGWLY(1)[0.58] TKFTDAGDTSFN(1)[0.75] EHNDFPMYTWRP(1)[0.92] WPDIEMDLGPLYA(1)[0.85] 6 Phage display (common panning) 3 PMID:17658545 Anti-Lpp20 monoclonal antibody L001 LPP20 lipoprotein Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of positive phage clones to the mAb L001 were measured by ELISA. The absorbance at 492 nm was measured and data shown were reproduced from the Fig. 5 in the reference. The phage library was served as the control, which had the value of 0.32 when binding to the mAb L001. In ELISA experiment, the peptide SWPLYSDASGLG appeared 11 times and the affinity value was presented as mean ± standard deviation, while VSLHDDASGTHR appeared 2 times and the two corresponding affinity values were separated by a slash. One mimotope (SWPLYSDASGLG) showed a good match with the Lpp20 proteins at 114–117aa (DASG) and the serum of mice induced by the phage clone clearly recognized Lpp20 protein. This mimotope provided an alternative approach for the diagnosis and development of a vaccine for H. pylori. 2803 QHYNIVNTQSRV[1.8 ± 0.08] 1 Phage display (common panning) 5 PMID:25070131 Anti-EGFR monoclonal antibody ICR-62 Epidermal growth factor receptor Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of mimotope conjugated with BSA (QHYNIVNTQSRV-BSA) to the mAb ICR-62 were measured by ELISA. The OD values at 450 nm were measured and data shown were reproduced from the Figure 2 in the reference. BSA was served as the control, which had the value of 0.26 ± 0.02 when binding to the mAb ICR-62. The affinity values were presented as mean ± standard deviation. Sequence analysis of eight reactive phages demonstrated that the interacting sequence QHYNIVNTQSRV plays a significant role in binding to anti-EGFR monoclonal antibody. 2804 GFPWHHHLVHAH[3.63] VRSHFRGHWVRF[2.81] SCYYPQLTRHRF[2.95] SSYYPQWPTDRF[4.27] SSYYPQLTAHRF[3.95] 5 Phage display (common panning) 4 PMID:25100324 Tumor necrosis factor Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA Absorbances were measured at 405 nm. The value shown is the binding fold for each phage clone, that is the ratio of its specific signal and the one generated by wild-type M13 phage. The real targrt is hexahistidine-tagged recombinant human TNFα (6His-rhTNFα). G6 peptide with the sequence of SSYYPQWPTDRF was screened and its binding to rhTNFα was investigated by isothermal titration calorimetry (ITC) experiments and molecular docking (MD) simulations. After pegylation, G6 peptide was conjugated on magnetic MPs and the system was assessed in the detection of rhTNFa in human serum. 2805 ETCRASCINESA(13/20)[1.22 ± 0.15] 1 Phage display (subtractive panning) 3 PMID:25324041 IgG from patients with CHB with HBsAg seroclearance following peg-IFN-α therapy Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The binding activities of positive phage clones to sera from chronic hepatitis B patients,with hepatitis B surface antigen (HBsAg) clearance, following pegylated interferon-α therapy were measured by phage ELISA. Absorbances were measured at 450 nm and data shown were reproduced from the Figure 1A in the reference. The original phage library was used as negative control, which had the value of 0.27 when binding to the sera. In ELISA experiment, the peptide ETCRASCINESA appeared 13 times and the affinity value was presented as mean ± standard deviation. To increase the screening specificity, microtiter wells were coated with purified sera IgG from the patients with CHB without HBsAg seroclearance, in order to absorb non-specific phages from the random phage peptide library. The unbound phages were then incubated with the sera IgG from patients with CHB, with HBsAg seroclearance, to screen for phage containing potential peptide biomarkers. In the validation phase, phage-ELISA results showed that the positive reaction rate of the ETCRASCINESA (named IFNC1) peptide phage clone was 92.0% with the HBsAg seroclearance group (n=50), which was significantly higher, as compared with the randomly selected HBsAg non-clearance group (12.0%, n=50) and the healthy control group (8.0%, n=50). The newly identified mimic peptide IFNC1 showed a high predictive validity HBsAg seroclearance in patients with CHB, following peg-IFN-α therapy. Therefore IFNC1 may be a potential serum biomarker, which could be used to predict the treatment outcomes of peg-IFN-α therapy. 2806 EWLFEFPTPVDA(12) DWIATWPDAVRS(12) 2 Phage display (subtractive panning) 2 PMID:25410289 Embryonic progenitor cell line SM30 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The library was subjected to cycles of selection on SM30 cells. To subtract nonspecific binding phages from the library, the phage library was pre-adsorbed against adult dermal fibroblasts cells before each of selection cycle. 2807 LTPPGRLSSWPL(2/17) SLSLVAPVLSLL(2/17) LTARQLVTNTTH(2/17) LTLPLNTKTVQD(1/17) QPPSINLTLYPM(1/17) LTAPLNVTMNPV(1/17) SVTLSLRLPFPS(1/17) MSNQQMRALSLL(1/17) PQSLQLPRFDMR(1/17) GSFLQLLPQPSP(1/17) FHPIKLAPYPAV(1/17) TYLNYSMGIDLE(1/17) STWPAFRLFTNI(1/17) HGLIEGVSNLVF(1/17) 14 Phage display (common panning) 5 PMID:25661733 Ovarian cancer cell lines OC-3 Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) To test the ability of these 14 phage clones to bind to other clear cell ovarian adenocarcinoma cells, the binding activity of each individual phage clone to two other clear cell ovarian cancer cell lines(ES2, SKOV-3) was further analyzed by flow cytometry. Overall, two phage clones, LTPPGRLSSWPL and SVTLSLRLPFPS, exhibited prominent binding to the ES2 and SKOV-3 cell lines. 2808 ALRINTDPDFTE(15/19) DARMWGHMTTML(1/19) VWAMENSIRMTL(1/19) KLFDPWLVQSHF(1/19) MHPNAGHGSLMR(1/19) 5 Phage display (common panning) 3 PMID:25728640 Polyclonal rabbit anti-AK IgG Arginine kinase (AK) Not determined. Not determined. Ph.D.-12 phage display library (X12) In the second and third round of biopanning, 50 ug/mL and 25 ug/mL IgG were incubated with amplified phages selected in the previous biopanning round, to enrich those phage displaying high-affinity binding. 2809 SHSLLHH(2) SIPSLNM(1) SQTQLPG(1) SQPDRNE(1) SSLFYKY(1) SPPGSPH(1) YPHLPEH(1) LSVLGDQ(1) LIRPSPD(1) DSSPLSL(1) ASAPLLG(1) QTSPPHI(1) GTPTPAI(1) NAPYGLR(1) EHFQPPL(1) TNDTSPS(1) VIKSWYQ(1) VESTGIW(1) VLHPSRN(1) 19 Phage display (subtractive panning) 1 PMID:25754556 Cyclin-dependent kinase inhibitor 2A, isoforms 1/2/3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Prior to p16 selection, non-specific binding phages were removed from phage display library by adsorption of 2ml phage library on blank plates three times. 2810 SHSLLHH(11) YAWDTYR(3) SLHQPHL(1) SHGNWWR(1) YRAPWPP(1) FPPSVIR(1) HAIYPRH(1) IPTHIRP(1) 8 Phage display (subtractive panning) 2 PMID:25754556 Cyclin-dependent kinase inhibitor 2A, isoforms 1/2/3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Prior to p16 selection, non-specific binding phages were removed from phage display library by adsorption of 2ml phage library on blank plates three times. 2811 SHSLLHH(13)[0.54] SLHQPHL(2)[0.57] YRAPWPP(2)[0.35] YAWDTYR(1)[0.36] FPPSVIR(1)[0.57] FPPLVLR(1)[NT] 6 Phage display (subtractive panning) 3 PMID:25754556 Cyclin-dependent kinase inhibitor 2A, isoforms 1/2/3 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Optical densities (ODs) of the enzymatic reaction were measured by ELISA plate reader with 405 nm filter set. ODs of the reaction in p16-coated wells were subtracted with the non-specific background signals to obtain the actual p16 binding signal of each phage clone. When the amount of phage display particles (PFU) was 6.25e10, the OD values were recorded. Data were reproduced from the graph. The OD value of wild-type phage is 0.13. Prior to selection, non-specific binding phages were removed from phage display library by adsorption of 2ml phage library on blank plates three times. 2812 GGSQGAY(2.1%) 1 Phage display (in vivo) 1 PMID:25762070 Soft tissue of prostate cancer xenograft Not determined. Not determined. Not determined. CX7C fUSE5 phage display library For round 1, tumor-bearing mice received 2e9 transducing units (TU) of phage peptide library i.v. After 24 h of circulation, the mice were perfused with 10 mM PBS (pH 7.4), and tumors were collected. Soft tissue and bonelike matrix compartments of the tumor were separated physically, weighed, and homogenized for phage recovery. The recovered phage pool was amplified and subjected to another round of selection. 2813 RIDAGTT(8.4%) PKRGFQD(4.2%) SPSQRQY(2.1%) GQVGIWS(2.1%) SGPTRGM(2.1%) GSQQQGR(2.1%) PGDQPRG(1.1%) 7 Phage display (in vivo) 2 PMID:25762070 Soft tissue of prostate cancer xenograft Not determined. Not determined. Not determined. CX7C fUSE5 phage display library For round 1, tumor-bearing mice received 2e9 transducing units (TU) of phage peptide library i.v. After 24 h of circulation, the mice were perfused with 10 mM PBS (pH 7.4), and tumors were collected. Soft tissue and bonelike matrix compartments of the tumor were separated physically, weighed, and homogenized for phage recovery. The recovered phage pool was amplified and subjected to another round of selection. The two peptides, PKRGFQD and SNTRVAP, targeted the cell surface of androgen-independent prostate cancer cells in vitro, and homed to androgen receptor-null prostate cancer in vivo. Purification of tumor homogenates by affinity chromatography on these peptides and subsequent mass spectrometry revealed a receptor for the peptide PKRGFQD, α-2-macroglobulin, and for SNTRVAP, 78-kDa glucose-regulated protein (GRP78). 2814 RIDAGTT(37.6%) SGPTRGM(33.3%) PKRGFQD(18.3%) PGDQPRG(3.2%) LDGPRAS(2.2%) 5 Phage display (in vivo) 3 PMID:25762070 Soft tissue of prostate cancer xenograft Not determined. Not determined. Not determined. CX7C fUSE5 phage display library For round 1, tumor-bearing mice received 2e9 transducing units (TU) of phage peptide library i.v. After 24 h of circulation, the mice were perfused with 10 mM PBS (pH 7.4), and tumors were collected. Soft tissue and bonelike matrix compartments of the tumor were separated physically, weighed, and homogenized for phage recovery. The recovered phage pool was amplified and subjected to another round of selection. 2815 SNTRVAP(33.7%) RLGLAWG(14.5%) SNNFVAP(3.6%) 3 Phage display (in vivo) 2 PMID:25762070 Bonelike matrix tissues of prostate cancer xenograft Not determined. Not determined. Not determined. CX7C fUSE5 phage display library For round 1, tumor-bearing mice received 2e9 transducing units (TU) of phage peptide library i.v. After 24 h of circulation, the mice were perfused with 10 mM PBS (pH 7.4), and tumors were collected. Soft tissue and bonelike matrix compartments of the tumor were separated physically, weighed, and homogenized for phage recovery. The recovered phage pool was amplified and subjected to another round of selection. The two peptides, PKRGFQD and SNTRVAP, targeted the cell surface of androgen-independent prostate cancer cells in vitro, and homed to androgen receptor-null prostate cancer in vivo. Purification of tumor homogenates by affinity chromatography on these peptides and subsequent mass spectrometry revealed a receptor for the peptide PKRGFQD, α-2-macroglobulin, and for SNTRVAP, 78-kDa glucose-regulated protein (GRP78). 2816 SNTRVAP(11.7%) GAGPASV(2.4%) SNTFVAP(2.4%) RLGLAWG(2.1%) VTRGVGF(2.1%) 5 Phage display (in vivo) 3 PMID:25762070 Bonelike matrix tissues of prostate cancer xenograft Not determined. Not determined. Not determined. CX7C fUSE5 phage display library For round 1, tumor-bearing mice received 2e9 transducing units (TU) of phage peptide library i.v. After 24 h of circulation, the mice were perfused with 10 mM PBS (pH 7.4), and tumors were collected. Soft tissue and bonelike matrix compartments of the tumor were separated physically, weighed, and homogenized for phage recovery. The recovered phage pool was amplified and subjected to another round of selection. The two peptides, PKRGFQD and SNTRVAP, targeted the cell surface of androgen-independent prostate cancer cells in vitro, and homed to androgen receptor-null prostate cancer in vivo. Purification of tumor homogenates by affinity chromatography on these peptides and subsequent mass spectrometry revealed a receptor for the peptide PKRGFQD, α-2-macroglobulin, and for SNTRVAP, 78-kDa glucose-regulated protein (GRP78). 2817 GSPREYTSYMPH SLSSYNGSALAS 2 Phage display (in vivo) 3-4 PMID:25768046 The ligated left carotid artery in male C57/BL6J mice Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) The ligated left carotid artery is the target for phage display, while the right artery serves as a normal flow control. Four days postligation, in vivo phage display was carried out. Phages were injected into the tail vein of the mice at a concentration of 2e11 plaque-forming units. After 2.5 h, mice were sacrificed by CO2 inhalation, and the carotid arteries were surgically isolated; phages were recovered from the vessels. Titration of phage normalized to the protein content of the corresponding LCA or RCA was used to determine the relative uptake between the target and control arteries. All four consensus clones showed a significantly increased binding to the targeted LCA. ACTPSFSKIC showed average enrichment of 7.3-fold; SLSSYNGSALAS showed enrichment of 4.9; ACNTGSPYEC showed enrichment of 6.1, and GSPREYTSYMPH showed enrichment of 4.4 in the target artery. 2818 CNTGSPYEC CTPSFSKIC 2 Phage display (in vivo) 3-4 PMID:25768046 The ligated left carotid artery in male C57/BL6J mice Not determined. Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) The ligated left carotid artery is the target for phage display, while the right artery serves as a normal flow control. Four days postligation, in vivo phage display was carried out. Phages were injected into the tail vein of the mice at a concentration of 2e11 plaque-forming units. After 2.5 h, mice were sacrificed by CO2 inhalation, and the carotid arteries were surgically isolated; phages were recovered from the vessels. Titration of phage normalized to the protein content of the corresponding LCA or RCA was used to determine the relative uptake between the target and control arteries. All four consensus clones showed a significantly increased binding to the targeted LCA. ACTPSFSKIC showed average enrichment of 7.3-fold; SLSSYNGSALAS showed enrichment of 4.9; ACNTGSPYEC showed enrichment of 6.1, and GSPREYTSYMPH showed enrichment of 4.4 in the target artery. 2819 WQRPSSW(2)[1.91, 1.80, 1.63] HLYWQRP(1)[1.28, 1.16, 0.98] 2 Phage display (competitive panning) 4 PMID:25771000 Human serum albumin Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA Selected 7-mer peptides were tested for binding human serum albumin (HSA), rabbit serum albumin (BuSA) and mouse serum albumin (MSA) by ELISA. The OD405 values were determined. Data shown were reproduced from graph. After removing unbound phage by repeatedly washing the plate with TBST, bound phage were eluted with 1ml HSA solution (100 mg/ml). 2820 ICARQDPAGNCS[0.50] 1 Phage display (common panning) 3 PMID:25779283 Polyclonal anti-LiAg IgGs L. infantum protein antigen (LiAg) Not determined. Not determined. LX-8 phage display library (XCX8CX) ELISA Plate was sensitized with e10 phages/mL of individual clone and Wild type phage (negative control). Positive and negative dog serum were employed. Binding was detected using a peroxidase conjugated anti-dog IgG antibody. Absorbance values at 492 nm were measured. The A492 values of the phage peptide binding to positive and negative dog serum were 0.50 and 0.08, while for wild type phage the values were 0.10 and 0.06, respectively. Peptide ICARQDPAGNCS has a ability to promote a state of immunity against L. infantum infection in murine model after immunization using liposomes as vaccine carrier and is a promising vaccine candidate against CVL. 2821 ETTTEPNRTRASAPA(5) HYNTEPKARDLRTIP(1) AYYAEPKQKTYLRTP(1) HDYTEPRFTYQISNA(1) WTEPKGPNAPQDPWM(1) GELETEPRKNAGIMP(1) GPAPDGTEPRYYSRS(1) ERTRTEPKIKSIVSM(1) VSTEPKRPYAPTTGC(1) SGSTEPKRLNWDNVS(1) SEAEPHVHRLIYRPL(1) SPAEPRNERTAARMR(1) AAEPRRHLSNLPDTR(1) SVPQLEPKRPKHRIY(1) SELEPRRPRSPYPVT(1) DRRQTEPITMKYTEP(1) FTTEPPKAGRTYQVW(1) GQTEPPRQISFKVAP(1) ARPRIEPEKLRQLPG(1) RLATEPTHNGSHPAN(1) GAQLEPNKATTARLT(1) TMPSKHHFLDRAHPP(1) RTASVPSRDRTNYAP(1) Y-TEP, TEP 23 Phage display (common panning) 3 PMID:10712615 Anti-human cardiac troponin I mAb 11E12 Human cardiac troponin I (hcTnI) Not determined. Not determined. f88.4-based X15 phage display library 2822 LTWGEMHTWTVQ(20) VSWPELYKWTWS(16) TTFDILDYWTSN(4) ITTSEIYNWRDT(3) ITNAELTNWNNG(2) GQPWTTWLESNT(2) ITAPELYAWFGS(1) LTMEELTRWSVY(1) ITLPELHAWKEN(1) ITIQEITAWPES(1) LTNQELLTWTAY(1) MDLAELSNWPHA(1) LSIADLYRWNTS(1) WQIWEYWPMDHN(1) VTLGELVSWPAE(1) LTLEELLFWKSP(1) LTRLELLEWDSP(1) LSWEELLRWASP(1) LTHTELLHWNGM(1) ITQADVWAWDTS(1) 20 Phage display (common panning) 3 PMID:25785734 Anti-HIV-1 monoclonal antibody VRC01 Human immunodeficiency virus type 1, HIV-1 Not determined. Not determined. Ph.D.-12 phage display library (X12) 2823 CEWSLWSFC(6) CSWTLLGYC(5) CSWNLMGFC(5) CNWEFWKYC(5) CNWEFWKYC(5) CSWNLMGFC(5) CPWVLHGFC(4) CTWTLLSFC(4) CSWSLNGFC(3) CTFTYWGFC(1) CPWYLMGYC(1) CNWSLLSFC(1) CEWTWFGYC(1) CLWSLTGFC(1) CQWTYYNFC(1) CEWRYWEYC(1) CDWLLHGFC(1) CPWMLSGFC(1) CTWSLSGFC(1) CLWMLEKFC(1) CTHSRAGSC(1) CSWSLLDFC(1) CIWEFLGFC(1) 23 Phage display (common panning) 3 PMID:25785734 Anti-HIV-1 monoclonal antibody VRC01 Human immunodeficiency virus type 1, HIV-1 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) 2824 DCFVRCRVACA(2)[NT] SCFALCRVPCQ(2)[NT] KCFTACRVDCF(1)[NT] SCFNRCRVPCF(1)[NT] SCFDRCRVGGY(1)[NT] QCFELCRVVCL(1)[NT] RCFNLCRVGCL(1)[NT] VCFPLCRVPCI(1)[NT] ACFKACRVNCA(1)[7.5 ± 3.2] ACFKHCRVACA(1)[10.6 ± 0.5] DCFQGCRVFCS(1)[NT] MCFDSCRVNCT(1)[112.7 ± 22.1] TCFHRCRVTCI(1)[NT] KCFQACRASCY(1)[NT] RCFEKCRAFCY(1)[NT] VCFSLCRAGCL(1)[NT] VCFGVCRALCA(1)[NT] VCFQLCRAQCI(1)[450 ± 75.5] LCWTGCRVSCF(1)[NT] GCWAPCRVGCY(1)[NT] RCWTACRGLCF(1)[NT] RCWQTCRVSCV(1)[NT] QCWRSCRVNCL(1)[NT] GCYTRCRVDCF(1)[NT] VCYALCRVSCD(1)[NT] DCYQLCRVSCE(1)[NT] TCFRSCKVACY(1)[NT] QCFQACKTLCW(1)[NT] 28 Phage display (subtractive panning) 2 PMID:23100545 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX3CX3CX phage display library Inhibition assay The bicyclic peptides inhibited hPK with IC50s ranging from 7.5 to 450 nM. Bicyclic peptides were chemically synthesized and their inhibitory activity (IC50, nM) is indicated. NT denotes not tested. The library was joined and subjected to a first affinity selection with hPK. The second round of panning was performed using neutravidin-coated magnetic beads to prevent the enrichment of streptavidin-specific peptides. 2825 VCGTGWCQLARCI(1) 1 Phage display (subtractive panning) 2 PMID:23100545 Plasma kallikrein (EC:3.4.21.34) Not determined. Not determined. Not determined. XCX4CX4CX phage display library The library was joined and subjected to a first affinity selection with hPK. The second round of panning was performed using neutravidin-coated magnetic beads to prevent the enrichment of streptavidin-specific peptides.