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

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‐t...
Source: Journal of Peptide Science - Category: Biochemistry Authors: Tags: Research Article Source Type: research