BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3384

Peptides:

Sequences
ASVQERK(2) AEKIQPR(2) TQKIQPR(1)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (common panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 30394584

Target Name:

Anti C-terminal appetite-regulating hormone[24-51] polyclonal antibody

Template Name:

C-terminal appetite-regulating hormone[24-51]Growth hormone secretagogue Growth hormone-releasing peptide Motilin-related peptide Protein M46

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:

Briefly, antibodies (3.4 to 1.7 μg; decreasing towards the third selection round) were incubated with phage libraries (2e11 pfu) in 100 μl of PBS solution with shaking at 400 rpm for 2 h in first selection round, 1 h in the second and for 0.5 h in the third. The resulting mixture was added to protein G or protein A coated magnetic beads (Dynabeads Protein G/A, Life Technologies, Invitrogen) in 1 mL of 0.1% BSA/0.1% PBST and incubated with shaking (400 rpm) to affinity capture the formed antibody-phage complexes. In each successive round of selection protein G and protein A beads were used alternatingly. Unbound phages were removed with repeated washings using 0.1 % PBST and bound phages were eluted with 400 μL of 200 mM glycine-HCl buffer (pH 2.2) containing 0.4 M NaCl and 0.1% BSA with 10 min of vigorous shaking (650 rpm). The beads were removed and the eluate was immediately neutralized by adding 1 M Tris-HCl (pH 9.1).

BiopanningDataSet Comments:

Curator:

Zhongjie Tang

Curation Date:

2019-01-29

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