Purification of immunoglobulin Y (IgY) from egg yolk by gel filtration chromatography

Abstract

Objective: Purification of immunoglobulin Y from egg yolk by chromatography is a new investigation. In this study, we developed an easy and inexpensive method for purification of IgY from egg yolk. Methods: The yolk from egg was separated and passed through a nylon mesh into a measuring cylinder. The egg yolk was diluted 10 times with HCL (3mM) and prepared suspension was centrifuged. Solid ammonium sulfate was added to 60% saturation. The precipitate was dissolved in phosphate buffer and applied on the gel-filtration columns (Sephadex G25, G200). Findings: The result of cellulose acetate electrophoresis at pH=9 showed some extra bands which were isolated with ammonium sulfate precipitation and sephadex G-25 chromatography. The purified band was observed in the Y–globulin region. After sephadex G-200 chromatography, SDS-PAGE electrophoresis of the product showed on band about 63KD. Conclusion: IgY extraction and purification with acid, centrifugation, ammonium sulfate and gel filtration are effective and simple operative methods.

Keywords

• immunoglobulin y (igy)
• gel filtration
• purification