BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3394

Peptides:

Sequences
FNKWMDCLSVTH

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (common panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 30417023

Target Name:

Anti-histamine H4 receptor (H4R) monoclonal antibody

Template Name:

Histamine H4 receptor, H4R

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 bingding affinity of the identified phages to human H4R mAb was examined by ELISA. The absorbance at 450 nm (OD450) was measured, which was reproduced from Figure 1 and shown.

Brief Description:

96-well microtiter plates were coated with affinity-purified anti-H4R antibody (40 μg/well) in sodium bicarbonate buffer (pH 8.6) and incubated overnight at 4°C. Blocking buffer consisting of 1% (w/v) BSA/Tris-buffered saline (TBS) was added to each well and incubated for 2 hr at room temperature, followed by 1 hr at 37°C using the phage library (total of 2e11 phages). The wells were washed six times with TBS and Tween 20 (TBST). Unbound phages were discarded and the wells were washed five times with TBST. Bound phages were eluted by adding elution buffer (0.2 M glycine-HCl in 1 mg/mL BSA [pH 2.2]) to each well, and the plates were gently rocked for 12 min at room temperature. The eluate was neutralized with 1 M Tris-HCl (pH 9.1). The eluate was amplified and precipitated with 20% (w/v) PEG-8000. A second round of panning was conducted using the first round of amplified eluate as the input phages. The entire screening protocol was repeated for a total of three rounds of panning.

BiopanningDataSet Comments:

The peptide FNKWMDCLSVTH, designated as P-FN12, was bound by H4R monoclonal antibody (mAb) with high affinity. Moreover, the P-FN12 + CTB@Lipo-formulated vaccine, used as nasal drops, decreased allergic symptoms such as sneezing and nasal rubbing in a rat model.

Curator:

Yuwei Zhou

Curation Date:

2019-01-29

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