BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 2357

Peptides:

Sequences
EPLQLKM(2) TALPPSY(2) AMISAIH(1) HVIANAG(1) GNTPSRA(1)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (in vivo)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

3-4

Reference:

PMID: 24320790

Target Name:

Four- to five-week-old mdx mice muscle

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

Ph.D.-7 phage display library (X7)

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

Four- to five-week-old mdx mice were injected intravenously with phages (one animal per round). Mice were perfused with phosphate-buffered saline (PBS) after 1 hour to remove unbound phages. The gastrocnemius and heart muscles were isolated and washed three times with Tris-buffered saline (TBS). Subsequently the organs were homogenized in 2 mL TBS with MagNA Lyser green beads (Roche Diagnostics) in the MagNA Lyser (Roche Diagnostics). The suspension was titrated and amplified. Up to four rounds of biopanning were performed for each fraction. After the third and fourth round, phages were plated and colonies were picked.

BiopanningDataSet Comments:

The selected peptides were synthesized with a FAM label and incubated with primary human control myotube cultures and analyzed with fluorescence microscopy. Clear fluorescence was observed for 7 of the 12 peptides. Based on the highest fluorescence intensity, P11 (HVIANAG) were chosen for in vivo evaluation. Mice were intramuscularly injected with 2.5 nmol of labeled peptide in the gastrocnemius muscle. Analyzed with fluorescence microscopy, P11 was found negative.

Curator:

Curation Date:

0000-00-00

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