BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 1574

Peptides:

Sequences
DSPQNPKTWKYI(1.00)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning, competitive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 16603084

Target Name:

Peptide TMGFTAPREPHY

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:

ELISA

Affinity Measurement Description:

The specificity of the interaction between the TMGFTAPREPHY and DSPQNPKTWKYI peptides was examined by ELISA. Phages displaying the DSP peptide specifically bind to a synthetic biotinylated TMG peptide. Conversely, phages displaying the TMG peptide bind to a synthetic biotinylated DSP peptide.

Brief Description:

The blocking, binding and washing strategies were carried out as instructed by the manufacturer with the following exceptions. Authors used (a) either 1% Bovine Serum Albumin (BSA) or casein for blocking nonspecific binding alternating these blocking substrates between subsequent rounds of bio-panning to prevent selection of phage recognising the blocking substances, (b) competitive elution with the bait peptide in rounds three and four and (c) bio-panning in solution for rounds two and four, using biotinylated peptide and a streptavidin matrix to prevent selection of plastic-binding phages. In the latter case, 10 μg of peptide were biotinylated using NHS-Biotin according to the manufacturer\'s instructions and incubated with phage particles as recommended by the manufacturer with either of the blocking reagents. The peptide bound phage particles were recovered either through a Streptavidin-agarose (Pierce Chemical Co. ILL) column or Streptavidin-Dynabeads (Dynal Co., Norway) followed by washing and competitive elution as described above. Following each round of bio-panning, the eluted phages were amplified to high titer according to supplier\'s instructions. The amplified fraction was used in the fourth round biopanning. The phage particles from the final fourth round eluate were plated at low density to allow isolation of single phage clones.

BiopanningDataSet Comments:

Curator:

Curation Date:

0000-00-00

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