BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 3360

Peptides:

Sequences	Affinity Values
KHRFNKD(2) PIISRAT(2) WSMRGTT(1) PGPSTTS(1) SKPLHKM(1) HRRPSRS(1) QISSLRA(1) WTDPHPP(1) HLAPAQY(1)	[1.62 ± 0.23] [0.15 ± 0.02] [NT] [NT] [NT] [NT] [NT] [NT] [NT]

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (competitive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID:

Target Name:

Anti-goat immunoglobulin G monoclonal antibody

Template Name:

Goat immunoglobulin G

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:

ELISA

Affinity Measurement Description:

The immunoplate and absorbance was estimated at 405 nm using a Synergy-HT microplate reader (Biotek, Winooski, USA). Each value was obtained by subtracting the value for the control from the absorbance value corresponding to each peptide. The control absorbance was determined using an immunoplate treated with blocking buffer only. The absorbance value was reproduced from Figure 2 and shown.

Brief Description:

Goat immunoglobulin G (IgG, 150 μg) was added to a culture dish with 1.5 mL TBST (0.1% Tween-20, 150 mM NaCl, 50 mM Tris-Cl, pH 7.5). The dish was incubated overnight at 4℃ and washed with TBST. Then, the dish was filled with blocking buffer (1 mM EDTA, 0.25% BSA in TBST), incubated at room temperature for 1 h, and washed with TBST. Rabbit anti-goat IgG antibody (27 μg) was added to the dish with 1 mL TBS (150 mM NaCl, 50 mM Tris-Cl, pH 7.5) and allowed to react at room temperature for 1 h. After washing, phages (2e11 pfu diluted in 1 mL TBS) were added to the dish and allowed to react at room temperature for 1 h. Rabbit anti-goat IgG antibody (27 μg in 1 mL TBS) was added to the dish and allowed to react at room temperature for 1 h for competitive elution of phages specific to rabbit antibody.

BiopanningDataSet Comments:

One of these sequences (KHRFNKD), which was rich in positive charge and homologous to the IgG-binding domains of Staphylococcal protein A, exhibited a high and specific affinity to rabbit antibody. This peptide was immobilized on a quartz crystal microbalance (QCM) biosensor, and the resulting QCM was found to be capable of capturing antibody and detecting antigen, with potential applications in antibody purification and enzyme immunoassay.

Curator:

Bifang He

Curation Date:

2019-01-29

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