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IgY-binding peptide screened from a random peptide library as a ligand for IgY purification.

Posted by on in 2017
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Khan KH1Himeno A1Kosugi S1Nakashima Y1Rafique A1Imamura A1Hatanaka T1Kato DI1Ito Y1. 2017. J Pept Sci. 23(10):790-797. doi: 10.1002/psc.3027. Epub 2017 Jul 31.
1
Graduate School of Science and Engineering, Kagoshima University, Kagoshima, 890-0065, Japan.

Abstract

Chicken egg yolk immunoglobulin (IgY) is a functional substitute for mammalian IgG for antigen detection. Traditional IgY purification methods involve multi-step procedures resulting in low purity and recovery of IgY. In this study, we developed a simple IgY purification system using IgY-specific peptides identified by T7 phage display technology. From disulfide-constrained random peptide libraries constructed on a T7 phage, we identified three specific binding clones (Y4-4, Y5-14, and Y5-55) through repeated biopanning. The synthetic peptides showed high binding specificity to IgY-Fc and moderate affinity for IgY-Fc (Kd : Y4-4 = 7.3 ± 0.2 μM and Y5-55 = 4.4 ± 0.1 μM) by surface plasmon resonance analysis. To evaluate the ability to purify IgY, we performed immunoprecipitation and affinity high-performance liquid chromatography using IgY-binding peptides; the result indicated that these peptides can be used as affinity ligands for IgY purification. We then used a peptide-conjugated column to purify IgY from egg yolks pre-treated using an optimized delipidation technique. Here, we report the construction of a cost-effective, one-step IgY purification system, with high purity and recovery. © 2017 The Authors. Journal of Peptide Science published by European Peptide Society and John Wiley & Sons Ltd.

KEYWORDS:

IgY purification; T7 phage display; chicken egg yolk; peptide library

PMID:
 
28758361
 
PMCID:
 
PMC5637892
 
DOI:
 
10.1002/psc.3027
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