3376 QVNGLGERSQQM(9) GLRTMQSPNSFY(1) VWDSGLRQSRPA(1) 3 Phage display (common panning) 4 PMID:30519000 Complement decay-accelerating factor Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Adherent HeLa cells were washed with serum-free RPMI-1640 and blocked with 16% culture medium containing 0.1% BSA for 1 hour, and then added to the stock solution of Ph.D.-12 phage peptide library (titer: 1e11 pfu/mL) for 1 hour. After washing on ice with a pre-cooled 0.1% PBST at 4°C to remove non-cell-bound phage, the phage bound to the cell surface was eluted on ice with a glycine buffer (pH 2.2) pre-cooled at 4°C immediately, and then added to a centrifuge tube pre-filled with 250 μL Tris buffer (pH 2.2). The effects of the peptide (QVNGLGERSQQM) on the proliferation and apoptosis of HeLa and SiHa cells were determined by Cell Counting Kit-8 (CCK-8), flow cytometry, and TUNEL assay, respectively, which showed that the peptide can bind to the CD55 molecule on the surface of cervical cancer HeLa and SiHa cells as a ligand peptide. It can also effectively inhibit the proliferation of cervical cancer cells and induce cell apoptosis. 3377 WLNMVWGVL(6) GRWAVLVNL(3) GVAVIFDFV(2) DTTVVVRMM(2) TPGGIVYRL(1) WSDSIVVHH(1) FQRFIVVAA(1) PGGLILLAD(1) SGWLILRDL(1) GDGCILVWR(1) TPGGILTWR(1) TPGGILFFR(1) ELGFIFLEV(1) GVPEIWMAF(1) GSVSIWYAD(1) GGWVIWLGL(1) TPGGIWLVE(1) PGGGIMSLV(1) LWIRIQMRH(1) TPGGITELY(1) TYRDITWHH(1) LKLLISDEH(1) FLNWISHHH(1) WFWFINEEH(1) LFTKIEHHH(1) RAVYIDDWL(1) ARRLIRYMT(1) GGERALFAS(1) FYYWAATRH(1) WFPYAWGVQ(1) TMELARFML(1) PGGQATYLE(1) WTGWYSCF(1) TWMPYFGG(1) GPDMGDTW(1) LDGELWFL(1) TPLLVWSHH(1) TPGGVWLRD(1) GGVGVWWSD(1) CPYSVWRPH(1) GVVEVWRAG(1) TPGGVYYRL(1) GVPGVYFVE(1) GKMVVYLQV(1) PYMDVYAHH(1) GFFRVYVEL(1) GGGGVFAMF(1) PLVDVFSSW(1) GVSTVFLSM(1) GGVLVFWSK(1) GGGAVGYSA(1) MDPGVGYWG(1) TPGGVGFWW(1) PGGLVAFNF(1) KYLAVAHHH(1) GVWCVAVLP(1) PGGRVLSFA(1) PGGRVLLVE(1) GGPTVLFGV(1) WAETVLFAH(1) SVSVVLWLH(1) LDYAVLVLY(1) TPGGVLELA(1) PGGEVLRLL(1) GWHLVPLVQ(1) EQYLVVESH(1) GSEVVMLLD(1) SPLWVMGHH(1) GGRVVMFAS(1) GYLTVNFST(1) EVDFVSFHH(1) GGAWVSYFG(1) GGTTVSQFL(1) FAAEVSHHH(1) PGGEVSLVL(1) GLSVVSAFM(1) GGLAVSAFL(1) VLLAVTLSE(1) PGGAVTFVF(1) YWGYVTHHH(1) PGGFVKAGM(1) CALAVKYHH(1) NGPAVHHHH(1) 83 Phage display (common panning) 8 PMID:30521603 Mast cell protease 2 Not determined. Not determined. Not determined. X9 T7 phage display library An aliquot of the amplified phages (~e9 pfu) were bound to 100 μl Ni-NTA beads by their His6-tags for 1 hr at 4°C under gentle agitation. Unbound phages were removed by washing ten times in 1.5 ml 1 M NaCl, 0.1% Tween-20 in PBS, pH 7.2, and two subsequent washes with 1.5 ml PBS. The beads were finally resuspended in 1 ml PBS. Activated mast cell protease 2 (~0.1 μg) was added to the resuspended beads and left to digest susceptible phage nonapeptides under gentle agitation at room temperature for 2 hours. PBS without protease was used as control. Phages with a random peptide that was susceptible to protease cleavage were released from the Ni-NTA matrix, and the supernatant containing these phages was recovered. To ensure that all of the released phages were recovered the beads were resuspended in 100 μl PBS (pH 7.2) and the supernatant, after mixing and centrifugation, was added to the first supernatant. To ensure that the His6-tags had been hydrolyzed on all phages recovered after protease digestion, 15 μl fresh Ni-NTA agarose beads were added to the combined phage supernatant and the mixture agitated for 15 min followed by centrifugation. 3378 GWSLFLEVL(3) GMATFLELH(3) AVLLFAGIL(3) WEVMFLDFH(2) MSLFWSDWH(2) WSVLFNWGH(2) TPGGYILML(2) GWGVFLDVG(1) GGVLFLDYF(1) GQALFLDIF(1) LIMLFLEEE(1) GIVGFLEVL(1) GWEFFLDAE(1) GVVSFLEMC(1) GWVSFLEPR(1) RATSFLEWH(1) VVMSFLDFS(1) GPVLWLEVM(1) AWVPWLDLV(1) GWVSYLEGG(1) GWMLYLEWR(1) LFVNFGEHH(1) WYVYFGELD(1) GATTFTDMV(1) GLETFIEVV(1) TPGGFVELY(1) WMTSFSDVM(1) WVVSFSETP(1) GIVVWSDVL(1) SVELWSEVL(1) AWVPWSEVH(1) VERFWSVEH(1) PVLLFMDFP(1) GGVLFMDEI(1) GWSAFMEFT(1) DTRFFQEWL(1) GFLSFWDLA(1) TWVSFFDLH(1) GVISFFEFV(1) GWVTWWDIG(1) EWMTYWEEG(1) EVIKWWDFH(1) GVVQYWGGD(1) WGVFYWEHH(1) GYALWWAIE(1) GWTVWFELV(1) SSTLYLIFE(1) YFWLWDHHH(1) WFLLWEPHH(1) GLWGWEGFR(1) TPGGFRGEG(1) RVYLWIGEH(1) EMELFWTHH(1) ECVVWGRHH(1) VAALWALFH(1) QGGKWALAY(1) ATYRWAASG(1) GPVVFLVRY(1) VLGIFLAVR(1) GSGQFLLFL(1) WAVWFLLIP(1) GYVTFLLIM(1) GWSTYLSYS(1) GAGFSTGW(1) VLYWFSSSW(1) LGITFMILH(1) LLWGFMFHH(1) VLVWWAKRA(1) GGNWWGVAT(1) GGLWWSYRS(1) TPGGWCQVQ(1) PGGDFRPAV(1) TPGGFRIVT(1) GGWVFRSLG(1) LFIFWHAVH(1) 75 Phage display (common panning) 5 PMID:30521603 Mast cell protease 1 Not determined. Not determined. Not determined. pComb M13 phage display library An aliquot of the amplified phages (~e9 pfu) were bound to 100 μl Ni-NTA beads by their His6-tags for 1 hr at 4°C under gentle agitation. Unbound phages were removed by washing ten times in 1.5 ml 1 M NaCl, 0.1% Tween-20 in PBS, pH 7.2, and two subsequent washes with 1.5 ml PBS. The beads were finally resuspended in 1 ml PBS. Activated mast cell protease 1 (~0.1 μg) was added to the resuspended beads and left to digest susceptible phage nonapeptides under gentle agitation at room temperature for 2 hours. PBS without protease was used as control. Phages with a random peptide that was susceptible to protease cleavage were released from the Ni-NTA matrix, and the supernatant containing these phages was recovered. To ensure that all of the released phages were recovered the beads were resuspended in 100 μl PBS (pH 7.2) and the supernatant, after mixing and centrifugation, was added to the first supernatant. To ensure that the His6-tags had been hydrolyzed on all phages recovered after protease digestion, 15 μl fresh Ni-NTA agarose beads were added to the combined phage supernatant and the mixture agitated for 15 min followed by centrifugation. 3379 LSFMPPE(2)[0.27] FSFIAPE(2)[n.a.] FSFLPPE(1)[0.57] FSFLAPE(1)[n.a.] FSFMVPE(1)[n.a.] LSFVPPE(1)[n.a.] FPWVPEL(1)[n.a.] FPWIPEE(1)[n.a.] [FL]SF[PAV]PE 8 Phage display (common panning) 3 PMID:30394584 Anti-GSFLSPEHQRVQ polyclonal antibody GSFLSPEHQRVQ Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The fraction of N-terminal specific antibodies obtained after purification was evaluated usingenzyme linked immunosorbent assay (ELISA). Biotinylated human ghrelin (Bachem, Bubendorf, Switzerland) was immobilized to streptavidin coated microtiter plates (StreptaWell, Roche, Basel, Switzerland) and affinity purified anti N-terminal antibodiesobtained from 5 µg of purified antibodies or unpurified (5 µg) were added. Binding was detected by secondary goat anti rabbit horseradish peroxidase (HRP)-conjugated antibodies (Merck Millipore, Darmstadt, Germany) diluted 1:5000 in 0.1% bovine serum albumin (BSA) and reagent 3,3',5,5'-tetramethylbenzidine (TMB) (Thermo Fisher Scientific, Waltham, MA, USA). The absorbance at 450 nm (A (405 nm)) was read using the Tecan Safire (Tecan, Grödig, Austria) microtiter plate reader. A (405 nm) values were preproduced from Figure 1B and shown. Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). Peptides were selected and tested using calcium screening assays. The most effective competitive antagonist FSFLPPE was further tested in vivo. Administration of the peptide produced no significant effect on either food intake or growth hormone (GH) release. Surprisingly, when co-administered with ghrelin the peptide significantly enhanced GH secretion and c-Fos expression. The evidence indicates that FSFLPPE might act as an ago-allosteric modulator. 3380 TVDDMAAMDGGH(1)[0.20] GLIDGMMFYQRG(1)[0.51] NIFASDTTKSTW(1)[0.31] WSQAYIAQFAGR(1)[n.a.] LPHRNLFEIEGR(1)[n.a.] 5 Phage display (common panning) 3 PMID:30394584 Anti N-terminal appetite-regulating hormone[24-51] polyclonal antibody N-terminal appetite-regulating hormone[24-51] Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The fraction of N-terminal specific antibodies obtained after purification was evaluated usingenzyme linked immunosorbent assay (ELISA). Biotinylated human ghrelin (Bachem, Bubendorf, Switzerland) was immobilized to streptavidin coated microtiter plates (StreptaWell, Roche, Basel, Switzerland) and affinity purified anti N-terminal antibodiesobtained from 5 µg of purified antibodies or unpurified (5 µg) were added. Binding was detected by secondary goat anti rabbit horseradish peroxidase (HRP)-conjugated antibodies (Merck Millipore, Darmstadt, Germany) diluted 1:5000 in 0.1% bovine serum albumin (BSA) and reagent 3,3',5,5'-tetramethylbenzidine (TMB) (Thermo Fisher Scientific, Waltham, MA, USA). The absorbance at 450 nm (A (405 nm)) was read using the Tecan Safire (Tecan, Grödig, Austria) microtiter plate reader. A (405 nm) values were preproduced from Figure 1B and shown. Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3381 NIFASAD(1)[0.31] GLVDVLF(1)[0.52] MNYREIY(1)[0.43] GLVDALY(1)[n.a.] MNRHIPA(1)[n.a.] GLFALERSK(1)[n.a.] 6 Phage display (common panning) 3 PMID:30394584 Anti N-terminal appetite-regulating hormone[24-51] polyclonal antibody N-terminal appetite-regulating hormone[24-51] Not determined. Not determined. Ph.D.-7 phage display library (X7) ELISA The fraction of N-terminal specific antibodies obtained after purification was evaluated usingenzyme linked immunosorbent assay (ELISA). Biotinylated human ghrelin (Bachem, Bubendorf, Switzerland) was immobilized to streptavidin coated microtiter plates (StreptaWell, Roche, Basel, Switzerland) and affinity purified anti N-terminal antibodiesobtained from 5 µg of purified antibodies or unpurified (5 µg) were added. Binding was detected by secondary goat anti rabbit horseradish peroxidase (HRP)-conjugated antibodies (Merck Millipore, Darmstadt, Germany) diluted 1:5000 in 0.1% bovine serum albumin (BSA) and reagent 3,3',5,5'-tetramethylbenzidine (TMB) (Thermo Fisher Scientific, Waltham, MA, USA). The absorbance at 450 nm (A (405 nm)) was read using the Tecan Safire (Tecan, Grödig, Austria) microtiter plate reader. A (405 nm) values were preproduced from Figure 1B and shown. Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3382 CIALNNNEC(1)[0.15] CSPKHQGSC(1)[0.15] CSPKHWGPC(1)[0.30] CIILNNNNC(1)[n.a.] 4 Phage display (common panning) 3 PMID:30394584 Anti N-terminal appetite-regulating hormone[24-51] polyclonal antibody N-terminal appetite-regulating hormone[24-51] Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA The fraction of N-terminal specific antibodies obtained after purification was evaluated usingenzyme linked immunosorbent assay (ELISA). Biotinylated human ghrelin (Bachem, Bubendorf, Switzerland) was immobilized to streptavidin coated microtiter plates (StreptaWell, Roche, Basel, Switzerland) and affinity purified anti N-terminal antibodiesobtained from 5 µg of purified antibodies or unpurified (5 µg) were added. Binding was detected by secondary goat anti rabbit horseradish peroxidase (HRP)-conjugated antibodies (Merck Millipore, Darmstadt, Germany) diluted 1:5000 in 0.1% bovine serum albumin (BSA) and reagent 3,3',5,5'-tetramethylbenzidine (TMB) (Thermo Fisher Scientific, Waltham, MA, USA). The absorbance at 450 nm (A (405 nm)) was read using the Tecan Safire (Tecan, Grödig, Austria) microtiter plate reader. A (405 nm) values were preproduced from Figure 1B and shown. Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3383 DERAYWFKSDLR(2) QIPERAYWVWSP(1) HALEDSKPPRTK(1) SAHNLKEQLKPP(1) 4 Phage display (common panning) 3 PMID:30394584 Anti C-terminal appetite-regulating hormone[24-51] polyclonal antibody C-terminal appetite-regulating hormone[24-51]Growth hormone secretagogue Growth hormone-releasing peptide Motilin-related peptide Protein M46 Not determined. Not determined. Ph.D.-12 phage display library (X12) Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3384 ASVQERK(2) AEKIQPR(2) TQKIQPR(1) 3 Phage display (common panning) 3 PMID:30394584 Anti C-terminal appetite-regulating hormone[24-51] polyclonal antibody C-terminal appetite-regulating hormone[24-51]Growth hormone secretagogue Growth hormone-releasing peptide Motilin-related peptide Protein M46 Not determined. Not determined. Ph.D.-7 phage display library (X7) Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3385 CKSDIQRSC(1) CIQKGHPMC(1) CIKFQPAQC(1) CVRYQPHQC(1) CVKLGKLYC(1) 5 Phage display (common panning) 3 PMID:30394584 Anti C-terminal appetite-regulating hormone[24-51] polyclonal antibody C-terminal appetite-regulating hormone[24-51]Growth hormone secretagogue Growth hormone-releasing peptide Motilin-related peptide Protein M46 Not determined. Not determined. Ph.D.-C7C phage display library (CX7C) ELISA Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1). 3386 SGVYKVAYDWQH(11) GLHTSATNLYLH(2) TVLSHPSTATLI(1) QQRPYVQDLRLI(1) 4 Phage display (subtractive panning) 4 PMID:30387183 BDNF/NT-3 growth factors receptor (EC:2.7.10.1)[1-430] Brain-derived neurotrophic factor, BDNF Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The phage particles were added to microtiter wells coated with 10 µg/mL of NTRK2, p75NTR (as nonspecific bindings), or bovineserum albumin (BSA; as a negative control) in coating buffer. The plate was washed three times after 1 hour incubation and 50µL of an anti-M13 antibody-horseradish peroxidase conjugate (GE Healthcare) was added. Finally, 50 µL of a substrate chromogen (TMB) was added and the plate was incubated at room temperature in the dark. Absorbance at 450nm was measured using Multiskan ELISA reader. The specificities and affinities of phage-displayed peptides were shown in Figure 3. In the first round, the Ph.D‐12 peptide library (New England Biolabs, Herts, UK) was added to 600 ng of recombinant BDNF/NT-3 growth factors receptor in 200 µL of tris‐buffered saline Tween 20 (TBS/T; 0.1% [vol/vol] Tween 20) and incubated for 2 hours. Then, 50 μL of protein G magnetic beads (Protein G Mag Sepharose; GE Healthcare UK Ltd, Little Chalfont, Buckinghamshire, UK) were added. After incubation and magnetic separation, the supernatant was removed (containing unbound phage). The magnetic beads were washed eight times with 1mL of TBS/T (0.1%) to remove weakly‐or nonspecifically‐bound phage. In the second round, one part of the amplified phage stock from the first round was preincubated with the beads in the absence of target to exclude protein G‐binding phage particles. Then, the unbound phage fraction was incubated with recombinant human BDNF/NT-3 growth factors receptor and specifically. The stringency of panning was increased in the third round of panning using reduced concentration of target protein and increased concentration of detergent. Also, a further round of biopanning was carried out and the phage particles were preincubated with recombinant human p75NTR ((nerve growth factor receptor/TNFRSF16 Fc Chimera; R&D System, Minneapolis, MN). The conserved residues of loop2 of brain-derived neurotrophic factor (BNDF) including S32‐GTVLKVPVQ‐L49 shared the most identity with the peptide 1 sequence (SGVYKVAYDWQH). Also, GLAM2SCAN algorithm, RRID:SCR_00178334 introduced peptide 1 as a motif due to the most frequency and similarity with BDNF. In silico molecular docking showed strong interactions between the peptide three dimensional models and the surface residues of BDNF/NT-3 growth factors receptor at the IgC2 domain. A consensus peptide sequence was then synthesized to generate a mimetic construct (named as RNYK). The affinity binding and function of this construct was confirmed by testing against the native structure of BDNF/NT-3 growth factors receptor in SH-SY5Y cells in vitro using flow-cytometry and MTT assays, respectively. RNYK at 5 ng/mL prevented neuronal degeneration of all trans-retinoic acid-treated SH-SY5Y with equal efficacy to or even better than BDNF at 50 ng/mL. 3387 NWTTLSRSVNWP WIYDTTRVIVPG HAMSPVFLSKYA LVPSILGATFIH DISASLQSNRWH AAGTFLMSMMSR NNLPTSRTLAGN GNNPLHVHHDKR HHLRIPYALDQT GTGAALAKVSEA IKPVRALYTLAD GTIRTSFWHTNT GVHSVFAPLTPN 13 Phage display (common panning) 3 PMID:30132552 Endoglin Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The absorbance was measured using a VICTOR X5 Multilabel plate reader (PerkinElmer, Inc., Singapore) at 490 nm, and the results are reported as the optical density. The binding activity of 16 monoclonal M13 phages to human recombinant CD105 protein and negative control blocking buffer was shown in Table II. The recombinant human CD105 protein (R&D Systems, Inc., Minneapolis, MN) at 100 µg/ml in sterile phosphate-buffered saline (PBS) was added to individual sterilised MaxiSorp plates (Thermo Fisher Scientific, Inc., Waltham, MA, USA) and incubated overnight at 4˚C. The plate was washed six times with Tris-buffered saline/0.1% Tween-20 (TBST) and then blocked for 1 h at 4˚C using 1% bovine serum albumin (BSA) in PBS. M13 phage display libraries were allowed to bind for 1 h at room temperature, and the unbound phages were washed away 10 times with TBST. Subsequently, the binding phages were eluted by an elution buffer, and the eluted phages were then amplified for another round of biopanning. A novel non-antibody-binding protein (WIYDTTRVIVPG) with a high affinity for recombinant human CD105 was identified by M13 phage biopanning. Fluorescence microscopy, flow cytometry and in vivo animal imaging were used to confirm the visualization effect of the novel peptide to the CD105-positive MNNG/HOS cell line in vivo and in vitro. Immunofluorescence of tissue sections was also used to identify the target efficiency of the peptide in tumour sections derived from an MNNG/HOS xenograft tumour model and osteosarcoma patients. This peptide specifically was found to bind to the CD105-positive osteosarcoma MNNG/HOS cell line and the osteosarcoma cells in the histological sections derived from an MNNG/HOS xenograft model and osteosarcoma patients in vitro. This peptide also successfully labelled an animal osteosarcoma xenograft model in vivo. Taken together, a novel peptide that can be used as a potent agent for CD105 molecular targeting was identified, and this peptide can be applied for osteosarcoma visualization in vitro and in vivo. 3388 ACLHFGVFCSMQHDNCG(3)[+++] ACLHFRDRCGSLQSGCG(2)[+] ACLHWTDSCLHYTQLCG(2)[+] ACMNYGVLCQFVTSTCG(2)[n.a.] ACLHYSHLCQTKMDACG(1)[n.a.] ACLHFAHSCEQQAMRCG(1)[n.a.] ACLHYAQSCDGQHSVCG(1)[n.a.] ACLHFQGMCSELMEGCG(1)[+] ACLHFGAHCSQVVTDCG(1)[n.a.] ACLHGRGECSDMDFQCG(1)[++] ACLHGRQGCSATTDNCG(1)[n.a.] ACLHGPGLCQGLSYPCG(1)[n.a.] ACLSFGTNCDSQVFTCG(1)[n.a.] ACFSYGQLCDRAVYKCG(1)[n.a.] ACLNYGQWCDRMSLGCG(1)[++] ACYHFGLHCSQTKILCG(1)[n.a.] ACFHYGTGCTREQLFCG(1)[n.a.] ACWHGPPGCDSTFYMCG(1)[n.a.] 18 Phage display (common panning) 3 PMID:30411004 Rabbit anti-goat IgG (H + L) superclonal secondary antibody Not determined. Not determined. Not determined. CX6CX6C phage display library Six individual selected sequences representing different sequence motifs were synthesized and functionalized with BiotinSH. As a negative control, a sequence (JK38) sharing no observable similarity to any of the chosen motifs was used. The BiotinSH-functionalized peptides were captured by streptavidin immobilized on magnetic beads. After incubation with the target protein and five washing cycles, each pulldown experiment was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The phage-encoded peptide library was selectively modified by the two-step modification under optimized conditions using NATBA as a linker and FlcSH as an additive, which was then subjected to three iterative rounds of affinity selection against the model target (rabbit anti-goat IgG (H + L) Superclonal Secondary Antibody). Briefly, biotinylated Rabbit anti-Goat IgG (H + L) Superclonal Secondary Antibody (ThermoFischer) was immobilized on magnetic streptavidin beads (Dynabeads M-280 Streptavidin, Invitrogen). The bead suspension and phage suspension were then mixed together and incubated for 30 min on a rotating wheel at room temperature. Second and third rounds of panning were performed following the same procedure but using in the second round instead of streptavidin beads neutravidin-coated magnetic beads. We were pleased to find strong consensus motifs within the resulting peptides sequences as aclear indication for target-specific binding. Most of the similarities are found in the N-terminal part of the first loop, consisting predominately of Leu, His, and Phe/Tyr. In the second loop, the highest conserved position is likewise the most N-terminal amino acid next to the central Cys, showing a strong preference for polaror acidic residues (Ser, Asp, Gln). None of the isolated sequences were found with an abundance higher than 3. 3389 YKLKIRTPQ YKLKVRTPQ YKLRIRTPQ YKLEIRTPQ YRLKIRTPQ YKLKIRTPR YKLKIQTPQ YKLKTRTPQ YELKIRTPQ YKLKIRAPQ YKLKIRIPQ HKLKIRTPQ NKLKIRTPQ YKPKIRTPQ YKLKIRTPL CKLKIRTPQ YKLKIRTLQ YKLKIRTSQ LELLKASRW LELLRASRW LELLEASRW LELLKVSRW LELLKAPRW LELLKALRW LESLKASRW LELPKASRW LGLLKASRW PELLKASRW LELLKASRR LQLLALSRT LRLLALSRT LQLLALSRA LQLLAPSRT PQLLALSRT LQLPALSRT LELVRRSPV LELARRSPV LELVRRSPA LELVHRSPV LELVRHSPV LELVRCSPV LEPVRRSPV LELVRRSSV LELVCRSPV LALIKRQPL LALLMRQRP LAMLTRGRP YKLKIRTPQ, LELLX(2)SRX 47 Phage display (common panning) 3 PMID:30373771 Ubiquitin carboxyl-terminal hydrolase 11 [1–244 or 24–244] Not determined. Not determined. Not determined. X9 PC89-based phage display library Proteins were immobilized on cobalt beads (Dynabeads) (solution phase panning). A total of 100 μg of USP11_DU was immobilized for each round. Bound phage were washed 10–20 times with PBS (10 mM phosphate buffer, pH 7.2–7.4, with 150 mM NaCl) plus 0.1% (v/v) Tween (PBST) and then PBS. Bound phage were eluted from the beads with 100 mM triethylamine and neutralized in 1 M Tris, pH 7.4. We discovered unique USP11-interacting peptide motifs. Isothermal titration calorimetry disclosed that the highest affinity peptides (AEGEFYKLKIRTPQ, KD of ~10 μM) exhibit exclusive selectivity for USP11 over USP4 and USP15 in vitro. Furthermore, a crystal structure of a USP11–peptide complex revealed a previously unknown binding site in USP11’s noncatalytic ubiquitin-like (UBL) region. This site interacted with a helical motif and is absent in USP4 and USP15. Reporter assays using USP11-WT versus a binding pocket–deficient double mutant disclosed that this binding site modulates USP11’s function in homologous recombination–mediated DNA repair. The highest affinity USP11 peptide binder fused to a cellular delivery sequence induced significant nuclear localization and cell cycle arrest in S phase, affecting the viability of different mammalian cell lines. The USP11 peptide ligands and the paralog-specific functional site in USP11 identified here provide a framework for the development of new biochemical tools and therapeutic agents. 3390 SDEEFNFINSAFDVADVNWYKNN[3.2] 1 Phage display (subtractive panning) 3 PMID:30203962 The variant of photoactive yellow protein (PYP) in the light-state Not determined. Not determined. Not determined. XDXXFNXINXAXXVXXVNXXKNX phage display library NMR spectroscopy All NMR experiments were carried out at CSICOMP (Department of Chemistry, University of Toronto) on an Agilent DD2 700 MHz spectrometer equipped with an HFCN cold probe. The Kd (μM) value was shown. Proteins were immobilized on neutravidin-coated plates. For selecting ligtht-state of cPYP binders, the phage libraries were depleted by allowing binding to dark-adapted protein (cPYP-Dark) for 1 h prior to transferring the unbound phage to the light-adapted plate for positive selection. The phage bound to the light-adapted protein (cPYP-Light) was washed 8 times with PT buffer, eluted and amplified overnight and the panning process was repeated 3 times. For panning rounds 2 and 3, a lower concentration of phage was used as the high-avidity of display often results in very high elute concentration. 3391 QDYLFNRINEAHFVNQVNVRKNV[20] 1 Phage display (subtractive panning) 3 PMID:30203962 The variant of AsLOV2 in the dark-state Not determined. Not determined. Not determined. XDXXFNXINXAXXVXXVNXXKNX phage display library NMR spectroscopy All NMR experiments were carried out at CSICOMP (Department of Chemistry, University of Toronto) on an Agilent DD2 700 MHz spectrometer equipped with an HFCN cold probe. The Kd (μM) value was shown. Proteins were immobilized on neutravidin-coated plates. For selecting dark-state of AsLOV2 binders, the phage libraries were depleted by allowing binding to light-adapted protein (AsLOV2-Light) for 1 h prior to transferring the unbound phage to the dark-adapted plate for positive selection. The phage bound to the dark-adapted protein (AsLOV2-Dark) was washed 8 times with PT buffer, eluted and amplified overnight and the panning process was repeated 3 times. For panning rounds 2 and 3, a lower concentration of phage was used as the high-avidity of display often results in very high elute concentration. 3392 QVDTQGENAVKV(189) PTVYHPELYQKA(187) AVMRQQTDELRL(186) AVHSNLFPGQPD(185) DPSDVLTLPFPR(183) FQFASGNEANET(181) WEIANPYWDGSE(170) VTVRENSPRKLA(166) YPNLLLLASVDV(166) QGVSDIHSRNLT(159) APAQPAESIHAY(155) RVTAPRPEFSTL(147) LPRVPPPVHSTT(143) ALSKTFEVAPLH(142) AYPSYLTSDGYH(141) IDTQYPSAMTLT(140) DIHRHVVGARTL(136) TTMRIAFHQLHT(134) RGELTNSGKARE(134) HGRFPLTSDVPT(123) SMPSMLFDTGED(121) ACAATPLNCGG(119) QIRDRIHDNELE(116) VETIPPLRYSDP(110) SENKNCNAGSLT(102) QPPHIHSALTLM(101) VAGTLPAPSPSY(90) NLGNYNDKEAVN(84) HDWSSKTETNAT(84) FMNTHDRADLSI(81) LLKHIEVSLPLA(80) QWYHRSDGGGSA(70) AINSTTGKRNVV(61) LACAVTGLICGG(59) RKAHQEKDSPRI(51) 35 Phage display (common panning) 1 PMID:30249752 Serum from patients with wasp sting injuries Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) A total of 4356 peptides were achieved in the 3-h group, and 4408 peptides in 4-day group. We compared the peptides between the two groups, and 35 specific peptides were achieved in the 4-day group. Amongst them, twelve peptide epitopes were matched with nine wasp venom proteins, namely, vitellogenin precursor, hexamerin 70b precursor, venom carboxylesterase-6 precursor, MRJP5, major royal jelly protein 8 precursor, venom acid phosphatase Acph-1 precursor, phospholipase A2, venom serine protease 34 precursor, and major royal jelly protein 9 precursor. The changes in serum IgM antibodies induced by wasp venom were confirmed by ELISA based on the 12 peptide epitopes. 3393 ADGVGDAESRTR[5.27e-8] 1 Phage display (common panning) 3 PMID:30260652 Chitosan Not determined. Not determined. Not determined. SUT12 M13 phage display library Surface plasmon resonance (SPR) Interactions between peptide and chitosan were quantified by SPR (Nicoya Lifescience, Waterloo, Canada). The binding constant (KD, M) was determined and shown. For biopanning of chitosan binding peptide, TBST (50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 0.1% Tween 20) was used to wash each well three times before adding 10 μl of the 12-amino acid-long random peptide library (SUT12, ~10^10 pfu) in 90 μl of TBST. The incubation time of the phage stock in the wells decreased with increasing rounds of selection. Following incubation at 25℃ for 4 h, the wells were washed for at least five times with TBST to remove unbound phages. In the first round, each well was washed for 6 times with no incubation. In the following rounds, the wells were washed twelve times with incubation for amounts of time. The bound phages were eluted by acid elution (50 μl of 50 mM glycine–HCl, pH 2.0). Then phosphate buffer (pH 7.5) was applied for neutralization of solutions. plates. Three rounds of biopanning were undertaken with chitosan according to the above mentioned methods. The chitosan-binding peptide (ADGVGDAESRTR) was used as the targeting moiety, followed by conjugation to the surface of poly(lactic-co-glycolic acid) nanoparticles as the drug carrier, which was then incubated with free chitosan. The noncovalently bound chitosan adheres to mucus layers and significantly enhances penetration of nanoparticles through the oral absorption barrier into circulation and then re-exposed the targeting ligand for later recognition of the fungal pathogen at the site of infection. 3394 FNKWMDCLSVTH 1 Phage display (common panning) 3 PMID:30417023 Anti-histamine H4 receptor (H4R) monoclonal antibody Histamine H4 receptor, H4R Not determined. Not determined. Ph.D.-12 phage display library (X12) ELISA The bingding affinity of the identified phages to human H4R mAb was examined by ELISA. The absorbance at 450 nm (OD450) was measured, which was reproduced from Figure 1 and shown. 96-well microtiter plates were coated with affinity-purified anti-H4R antibody (40 μg/well) in sodium bicarbonate buffer (pH 8.6) and incubated overnight at 4°C. Blocking buffer consisting of 1% (w/v) BSA/Tris-buffered saline (TBS) was added to each well and incubated for 2 hr at room temperature, followed by 1 hr at 37°C using the phage library (total of 2e11 phages). The wells were washed six times with TBS and Tween 20 (TBST). Unbound phages were discarded and the wells were washed five times with TBST. Bound phages were eluted by adding elution buffer (0.2 M glycine-HCl in 1 mg/mL BSA [pH 2.2]) to each well, and the plates were gently rocked for 12 min at room temperature. The eluate was neutralized with 1 M Tris-HCl (pH 9.1). The eluate was amplified and precipitated with 20% (w/v) PEG-8000. A second round of panning was conducted using the first round of amplified eluate as the input phages. The entire screening protocol was repeated for a total of three rounds of panning. The peptide FNKWMDCLSVTH, designated as P-FN12, was bound by H4R monoclonal antibody (mAb) with high affinity. Moreover, the P-FN12 + CTB@Lipo-formulated vaccine, used as nasal drops, decreased allergic symptoms such as sneezing and nasal rubbing in a rat model. 3395 KRLTPLSPDHYD(14/30)[4.8 ± 1.0] QLKLHNKGLISS(1/30)[3.2 ± 1.5] GWSSREASQEGN(1/30)[NT] NGVFAMLFPMQS(1/30)[NT] YGILARDNDSLG(1/30)[NT] VAYEGSYFAVYQ(1/30)[NT] SIDSGYRPGVRY(1/30)[NT] ADHWWSYAYQKP(1/30)[NT] VVVSSLSAPYKD(1/30)[NT] VPAPPISSASD(1/30)[NT] ITTPDWELMRGM(1/30)[NT] GIVEPSDRSTRG(1/30)[NT] AFDSPDNPVASN(1/30)[NT] EYHDYESANAAD(1/30)[NT] 14 Phage display (subtractive panning) 3 PMID:30344835 Ovarian cancer tissue slice Not determined. Not determined. Not determined. Ph.D.-12 phage display library (X12) Flow Cytometry Peptides were synthesized (>98% purity) and conjugated with FAM (excitation wavelength = 490 nm, emission wavelength = 525 nm) at the 5'- and carboxyl termini (Protech Co., Ltd., Taiwan). These fluorescently-labeled peptides were then used for fluorescence microscopy analysis and flow cytometric assays (BD Accuri™ C6, Becton, Dickinson and Company, USA). The resulting fluorescence intensity of the affinity reagents was further used to calculate the equilibrium dissociation constant (Kd, nM) with Prism software (GraphPad Software, USA). Selection was performed against ovarian cancer tissue slides by the integrated microfluidic system capable of automating SELEX and phage display technology. Briefly, an M13 phage library was incubated with normal tissues (“negative selection” step), and then the unbound M13 phages were incubated with cancer tissues (“positive selection” step). There were three rounds of both positive and negative panning involved for the entire screening process. 3396 GTGSQAS(7)[2.83] WTTSVGT(1)[NT] TVNFKLY(1)[NT] GVEGHKP(1)[NT] FTAERYY(1)[NT] QQTNWSL(1)[NT] 6 Phage display (common panning) 5 PMID:30592604 Gold Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) Surface plasmon resonance (SPR) To evaluate the binding affinity of the peptide GTGSQAS against the gold surface, the SPR analysis was performed. The binding affinities (Kd, μM) of all peptides were determined. The Kd values of previously reported gold-binding peptides VSGSSPDS and KHKHWHWG are 2.28 μM and 3.67 μM, respectively. The biopanning process was performed using the gold-patterned microfluidic chip with NEB Ph.D.-7 phage display peptide library kit. Ten microliters of phage solution (2e12 pfu mL−1) in 0.1 vol % PBST (PBS (pH 7.4) containing 0.1 vol % Tween 20) was loaded into the microfluidic biopanning platform at a flow rate of 1 μL min−1 at room temperature. Then PBST (0.1 vol % Tween 20 in the first round and 0.5 vol % Tween 20 from the second round) was introduced at 0.5 mL min−1 for 30 min to remove nonspecifically or weakly bound phages. The remaining high-affinity phages were eluted by injecting an elution buffer (0.2 M glycine-HCl (pH 2.2) with 1 mg mL−1 BSA) at 0.1 mL min−1 for 10 min, and the eluate was neutralized with 150 μL of 1 M Tris-HCl (pH 9.1). Five rounds of biopanning were performed, and the eluted phages from each round were amplified by incubation with ER2738 host cells. Finally, the phage peptide sequence was analyzed by sequencing the phage plasmid DNA. The surface plasmon resonance analysis shows that the binding affinity of the identified GTGSQAS is comparable to previously reported gold-binding peptides (GBPs). Moreover, molecular dynamics simulations are performed to understand its binding affinity against the gold surface in detail. Theoretical calculations suggest that the association and dissociation rates of the GBPs depend on their sequence-dependent conformations and interactions with the gold surface. 3397 GTGSQAS(4) GVEGHKP(3) QLAVAPS(2) MNSNIPI(2) TVNFKLY(2) ANPKNFS(1) FSGGGNH(1) ELWSLYD(1) NLQPPAY(1) RHFIKEL(1) NDLMNRA(1) 11 Phage display (common panning) 4 PMID:30592604 Gold Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The biopanning process was performed using the gold-patterned microfluidic chip with NEB Ph.D.-7 phage display peptide library kit. Ten microliters of phage solution (2e12 pfu mL−1) in 0.1 vol % PBST (PBS (pH 7.4) containing 0.1 vol % Tween 20) was loaded into the microfluidic biopanning platform at a flow rate of 1 μL min−1 at room temperature. Then PBST (0.1 vol % Tween 20 in the first round and 0.5 vol % Tween 20 from the second round) was introduced at 0.5 mL min−1 for 30 min to remove nonspecifically or weakly bound phages. The remaining high-affinity phages were eluted by injecting an elution buffer (0.2 M glycine-HCl (pH 2.2) with 1 mg mL−1 BSA) at 0.1 mL min−1 for 10 min, and the eluate was neutralized with 150 μL of 1 M Tris-HCl (pH 9.1). As the rounds progress, the proportion of high-affinity bacteriophages increases and fewer bacteriophages are removed from the surface after the washing step. Thus, the number of eluted bacteriophages increases as the rounds continue. 3398 HAAWHFI(2) VSRDTPQ(2) GTGSQAS(1) GASESYL(1) HSHTLTW(1) NHFTLNQ(1) YNGSANQ(1) SSQSLRE(1) TVNFKLY(1) QQTNWSL(1) YSEPAVT(1) SYTDLLR(1) GQSEKHL(1) TIVGAKD(1) QLYREFN(1) SHVNVPS(1) TTANVRI(1) ESRVMSR(1) FSGGGNH(1) NQIYSAN(1) TTQVLEA(1) ETALIAA(1) TSQYLMI(1) 23 Phage display (common panning) 3 PMID:30592604 Gold Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The biopanning process was performed using the gold-patterned microfluidic chip with NEB Ph.D.-7 phage display peptide library kit. Ten microliters of phage solution (2e12 pfu mL−1) in 0.1 vol % PBST (PBS (pH 7.4) containing 0.1 vol % Tween 20) was loaded into the microfluidic biopanning platform at a flow rate of 1 μL min−1 at room temperature. Then PBST (0.1 vol % Tween 20 in the first round and 0.5 vol % Tween 20 from the second round) was introduced at 0.5 mL min−1 for 30 min to remove nonspecifically or weakly bound phages. The remaining high-affinity phages were eluted by injecting an elution buffer (0.2 M glycine-HCl (pH 2.2) with 1 mg mL−1 BSA) at 0.1 mL min−1 for 10 min, and the eluate was neutralized with 150 μL of 1 M Tris-HCl (pH 9.1). As the rounds progress, the proportion of high-affinity bacteriophages increases and fewer bacteriophages are removed from the surface after the washing step. Thus, the number of eluted bacteriophages increases as the rounds continue. 3399 TWLPLSL(1) VTWLPNI(1) TWVSGTV(1) SFAGTAF(1) ANDDRLV(1) ANPGVWL(1) SWTALGP(1) SHVNVPS(1) AHINVPS(1) VSPSYII(1) VVISDRY(1) EYWQDKP(1) LNKVAVP(1) DHITGYS(1) HTAITLT(1) YPQTFAT(1) HSGYIRF(1) TPFYGSW(1) HSHTLTW(1) NGAMVGV(1) HAAWHFI(1) QLAVAPS(1) ETALIAA(1) QLYREFN(1) 24 Phage display (common panning) 2 PMID:30592604 Gold Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) The biopanning process was performed using the gold-patterned microfluidic chip with NEB Ph.D.-7 phage display peptide library kit. Ten microliters of phage solution (2e12 pfu mL−1) in 0.1 vol % PBST (PBS (pH 7.4) containing 0.1 vol % Tween 20) was loaded into the microfluidic biopanning platform at a flow rate of 1 μL min−1 at room temperature. Then PBST (0.1 vol % Tween 20 in the first round and 0.5 vol % Tween 20 from the second round) was introduced at 0.5 mL min−1 for 30 min to remove nonspecifically or weakly bound phages. The remaining high-affinity phages were eluted by injecting an elution buffer (0.2 M glycine-HCl (pH 2.2) with 1 mg mL−1 BSA) at 0.1 mL min−1 for 10 min, and the eluate was neutralized with 150 μL of 1 M Tris-HCl (pH 9.1). In the first round, a washing buffer containing a lower concentration of surfactant(0.1 vol % PBST) was used, which allows a higher diversity in the eluted phage pool and preserves more potential high-affinity bacteriophages.The surfactant concentration was then increased to 0.5 vol % PBST from the second round so that only phages having high affinity to the target remain. 3400 GVEGHKP(7) MVGKPQP(1) QTTSVGT(1) YNGSANQ(1) HGGVRLY(1) FSGGGNH(1) 6 Phage display (common panning) 5 PMID:30592604 Not determined. Not determined. Not determined. Ph.D.-7 phage display library (X7) A control experiment was also performed using another microfluidic device with the same initial phage display library(Ph.D.-7 phage display peptide library), which has no gold pattern on the surface, to exclude the possibility that the identified peptide was obtained by the interaction with PDMS, glass, or PEG coating.