از زرده تخم مرغ با روش كروماتوگرافي ژل فيلتراسيون Y خالص سازي ايمونوگلوبولين

PURIFICATION OF IMMUNOGLOBULIN Y (IGY) FROM EGG YOLK BY GEL FILTRATION CHROMATOGRAPHY

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 one band about 63KD. CONCLUSION: IgY extraction and purification with acid, centrifugation, ammonium sulfate and gel filtration are effective and simple operative methods.