BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3630

Peptides:

Sequences
RRPPR(5/24)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (common panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 34111267

Target Name:

Rat heart microvascular endothelial cell, RHMVEC

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Novagen T7 X7 phage display library

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

Rat heart microvascular endothelial cell (RHMVEC) (80% confluent; approximately 2.0e7 cells / 100 mm dish) were washed with PBS and preincubated in serum-free medium at 37°C for 30 minutes and inoculated with an extract (5.0e9 pfu) of the T7 phage library to reach a multiplicity of infection (MOI) of 250. After incubation for 1 hour at 37°C, cells were washed with ice-cold PBS and acid washed with 0.1N HCl pH 2.2 for 15 seconds to remove unbounded and weakly associated phages from the cell surface. Cells were then trypsinized, centrifuged, and lysed with sterile deionized water on ice and internalized phages were amplified. After completion of 6 rounds of selection/amplification, Escherichia Coli BL21 was infected with the resulting phages and plated, individual plaques were picked, amplified, and sequenced.

BiopanningDataSet Comments:

Phage display technology was used to identify a small peptide (RRPPR) that was internalized into endothelial cells. This peptide was potent in blocking nitric oxide (NO) release. Fusing RRPPR with a minimal Cav inhibitory domain (CVX51401) dose-dependently blocked NO release, VEGF induced permeability, and retinal damage in a model of uveitis.

Curator:

Bowen Li

Curation Date:

2023-10-13

First |

Prev |

3630

| Next

| Last