Journal of Peptide Science
Journal of Peptide Science
5 years ago

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

IgY-binding peptide screened from a random peptide library as a ligand for IgY purification
Takaaki Hatanaka, Yosuke Nakashima, Arisa Himeno, Yuji Ito, Ayana Imamura, Shouhei Kosugi, Abdur Rafique, Dai-Ichiro Kato, Kamrul Hasan Khan
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. From the random peptide library constructed on T7 phage display system, chicken egg yolk immunoglobulin (IgY)-binding peptides were successfully isolated. Those peptides showed high moderate affinity and specificities to IgY. Finally, IgY binding peptide-conjugated affinity column was prepared to recover IgY with high purity (~92%) from pre-treated egg yolk.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/psc.3027

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