BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3273

Peptides:

Sequences

CIIGSYVC
CNLSVPAC
CLSPIGEC
CLGKGSVC
CGRDSKQC
CAAAPIRC
CAGGGAYC
CMRGKGLC
CASGSENC
CVPMRLQC
CGRAKVRC
CLPISSSC
CMARQARC
CRECGERC
CAGKSSNC
CIRREKRC
CNIRRQGC
CPQPRPLC
CLANLQTC
CAVVFSQC
CSIRRESC
CWRSVEVC
CADVLRPC
CEGESASC

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 11689892

Target Name:

Human endothelial cells stimulated with vascular endothelial growth factor (VEGF)

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

CX6C phage display library

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

A method termed biopanning and rapid analysis of selective interactive ligands (BRASIL) was used. BRASIL allows separation of phage-cell complexes from the remaining unbound phage; this is accomplished by a differential centrifugation that drives the cells from a hydrophilic environment into a non-miscible organic phase. Because the organic phase is hydrophobic, it excludes water-soluble materials surrounding cell surfaces. Bound phage are recovered from the cell pellet whereas the unbound phage remain soluble in the upper aqueous phase, eliminating the need for repeated washes. A two-step panning strategy to isolate phage that bind to angiogenic ECs was used. First, to decrease non-specific binding, we pre-cleared the phage library on starved ECs (before panning on the same cell line stimulated with recombinant VEGF165); starved human umbilical vein ECs (HUVECs) were incubated with the phage library and centrifuged through the organic phase. Second, the unbound phage pool left in the aqueous phase was transferred to a fresh tube and incubated with VEGF165-stimulated HUVECs. After centrifugation through the organic phase, phage bound to the VEGF165-stimulated HUVEC pellet were recovered by bacterial infection, amplified and subjected to two more rounds of selection.

BiopanningDataSet Comments:

The motif PQPRPL is a novel chimeric ligand mimic that binds specifically to VEGF receptor-1 and to neuropilin-1.

Curator:

Lixu Jiang

Curation Date:

2018-01-19

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