BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3437

Peptides:

Sequences

Affinity Values

GLHTSATNLYLH(13)
SGVYKVAYDWQH(6)
FIPFDPMSMRWE(3)
ALAVAPSRWWNE(3)
KASGSPSGFWPS(3)
NASSFPTNSRWA(2)
QFDYMRPANDTH(2)
GQSEHHMRVASF(2)
GTGLVTLPRLTV(2)
DLILLNGGSQQA(1)
LPVGNQSDVHFD(1)
SPLCCNSGTDQT(1)
DPIKHLEDQAST(1)
AAIEPSAGLAYF(1)
SPYTWGSGDWRS(1)
DQFVHDVKGTKH(1)
AHWGMSARTSAW(1)
TNRTHNINALHF(1)
GNHNNSVTTWHL(1)
MHPNAGHGSLMR(1)
TPGGLLRNDRPL(1)
NIYTTPWGSNWS(1)
HLARTTDMLHPY(1)
QMSPGSTARSAL(1)
TVQSLPPHSPYD(1)
MDNKTTRWRDPL(1)
HSIALTTTLRTP(1)
MSKSTMSLSSNA(1)
TSIQISNAHPKS(1)
HTSYGTTWSESR(1)
NHLSTPVWSITG(1)
WSMNHTNLSYTS(1)
QPDFTPRPAYSR(1)
ITGCDPTKQTQG(1)
AYPQKFNNNFMS(1)
YDIHWSPDSRLK(1)
ASATVSKPWPTG(1)
KVMGLSVTSHAE(1)
SNSSLELPNLNS(1)
YIWQGPGSAGTP(1)
HGNPRSPNQFST(1)
QCFNSVCLHTNP(1)
QQHPLITRLISH(1)
ALEVNGMRPAWA(1)
HRGDTTYNHLHP(1)
SQGLSLSTMVTR(1)
ADPLGISYLRSS(1)
KATDFPDGRLNY(1)
SAELPHKTGSVL(1)
GSAPLLTVDTSK(1)
STPIFAEATARS(1)
LPHLPSQNVRAP(1)
SCDMQERCLAGL(1)
VLYEQVLAAPME(1)
TAGAIADFQSLT(1)
LTHYPVTGSTGS(1)
SEFQSTHWLDSL(1)
WIRPPSGPMYSF(1)
DPTRQEPHLFLL(1)
QAMMWLPVALAA(1)
SGKLLMLPVVSR(1)
YTTIRWSEFSAW(1)
DRWVARDPASIF(1)
ISSTDRLLAAGL(1)
SGTSLLHYHSYH(1)
DSQFNKYSIATV(1)
SNLSARNTISHG(1)
MDGSSREVNDAV(1)

[12.6]
[5.7]
[7.2]
[76.3]
[NA]
[20.6]
[3.3]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]
[NA]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (common panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 29956504

Target Name:

Human breast cancer cell line MCF7

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-12 phage display library (X12)

Affinity Measurement Method:

Flow Cytometry

Affinity Measurement Description:

The fluorescence intensity of each FITC-labeled peptide at 1.0e–6 m (also defined as the fluorescence per unit, FPU) was measured by a spectrofluorometer (VARIAN, Cary Eclipse, USA, Ex = 488 nm, Em = 535 nm). The FPU was used for calibration of the relative affinity. The mean fluorescence intensity (MFI) of 10000 cells was measured by a FACSCalibur flow cytometer (Becton Dickinson, USA). MCF7 cells incubated with blank RPMI-1640 medium were taken as the background group and MCF7 cells incubated with FITC-AGP were used as the control group. The relative affinity of a peptide to MCF7 cells was calculated using the following formula: Relative affinity = ((MFIpeptide -MFIbackground)/FPUpeptide)/((MFIcontrol - MFIbackground)/FPUcontrol), which means the affinity ratio of the given peptide to the control peptide. The mean relative affinity of ligand peptides to MCF7 cells was shown in Figure 2A and shown.

Brief Description:

MCF7 cells were seeded in a 25 cm2 flask and cultured until confluence (about 3 .0e6 cells per flask). Then the cells were rinsed twice with PBS and blocked with 0.5% bovine serum albumin in RPMI-1640 medium (BSA-1640) at 4 °C for 30 min. After blocking, the cells were coincubated with phages (1.0e11 pfu, Ph.D.-12 peptide library kit, the input phage library, displaying a diversity of 1.0e9 dodecapeptides) in BSA-1640 at 4 °C for 30 min to allow for binding or internalization between phages and cells. The medium containing unbound phages was discarded and the cells were washed ten times with BSA-1640 containing 0.1% Tween-20 to remove weakly bound phages. Afterward, the phages that bound strongly to MCF7 cells were eluted down with Glycine-HCl (2 m, pH 2.2) solution and the acidic eluate was recovered as one output phage sublibrary, which had higher cell-binding affinity to MCF7 cells than the input phage library. Finally, after washing thrice, the cells were lysed by Tris-EDTA solution (50e−3 m Tris, 1.0e−3 m EDTA, 150e−3 m NaCl, 2% deoxycholate, pH 10.0) and the cell lysate containing intracellular phages was recovered as another output phage sublibrary, which had higher cell-internalizing efficacy to MCF7 cells than the input phage library. The phage titer was assayed by counting plague forming units. The recovery rate of phage in each round was calculated as the ratio of output to input. The two output phage sublibraries (eluate and lysate) were amplified and used as the input phage libraries of the next panning round, respectively. A total of four rounds were performed to achieve phage sublibraries with sufficient affinity to MCF7 cells.

BiopanningDataSet Comments:

Four of these ligand peptides [FIPFDPMSMRWE (FIP), NASSFPTNSRWA (NAS), GLHTSATNLYLH (GLH), and ALAVAPSRWWNE (ALA), respectively] exhibit high affinity to MCF7 human breast cancer cells. Among them, NAS and ALA are reported for the first time, whose affinities are 20.6 and 76.3 times that of the random peptide control, respectively. Both NAS and ALA modifications to doxorubicin-loaded lipid nanosytems [LP(DOX)] show stronger tumor inhibition, longer animal survival time, and less body weight loss, compared to unmodified or control peptide modified nanosystems, on an MCF7 tumor-bearing mouse model.

Curator:

Bifang He

Curation Date:

2019-05-28

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