BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3462

Peptides:

Sequences	Affinity Values
RLCSWISPCSA(4) FGCSWLFPCPF(2) RLCSWVSPCSA(1)	[6] [8] [10]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (common panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 15233528

Target Name:

B-domain deleted form of human factor VIII (BDDrFVIII)

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

TN7/1 phage display library

Affinity Measurement Method:

ELISA

Affinity Measurement Description:

Upon completion of the second and third rounds of selection, randomly-chosen individual phage isolates from pools generated by both elution conditions were screened for binding to BDDrFVIII by indirect phage ELISA. Background binding levels were established using wells coated with streptavidin alone or with BSA. Phage isolates exhibiting ELISA signals significantly above background (A630 > 0.25) were scored as positive for binding to BDDrFVIII. The ratio of the ELISA signal with target to the ELISA signal without target was shown.

Brief Description:

Biotinylated BDDrFVIII protein that was confirmed to be active after derivatization was immobilized onto streptavidin-coated magnetic beads and used as a target in six separate selection campaigns. Each campaign incorporated three rounds of selection from a single library, using one elution condition for phage bound to the immobilized BDDrFVIII. A round of selection consisted of a binding step in which the phage library was first incubated in suspension with the target, followed by a plate wash step to remove unbound or weakly-bound members of the phage library, and an elution step to recover a pool of phage. This results in a population of phage isolates enriched in members that bind the target under binding and wash conditions and release the target under elution conditions. Finally, elution pools from each round of selection were separately amplified by infection and growth in E. coli cultures. The purified phage preparations from this amplification step served as the input for subsequent rounds of selection.

BiopanningDataSet Comments:

The consensus sequence is rlCSWѲsPCsa (uppercase letters stronger than lowercase, Ѳ = V, I, L, F).

Curator:

Ying Wang

Curation Date:

2019-08-17

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