p53蛋白的原核表达及其鸡卵黄抗体的制备

目的 制备抗p53蛋白卵黄抗体。方法 原核表达p53融合蛋白，纯化后免疫产蛋母鸡。结果 融合蛋白能有效刺激母鸡产生抗体，末次免疫后其卵黄抗体滴度达到1：10^4,并且能维持较长时间。结论 本文介绍的方法是一种简便、经济、特异、高效、均一、来源充足的抗体产生方法。

Preparation of egg yolk immunoglobulin (IgY) against p53 protein expressed in E.coli]

OBJECTIVE: To isolate immunoglobulin (IgY) from egg yolk of p53 protein-immunized hens and to study its reactivity to the antigens. METHOD: Immunization of egg-laying hens was performed for 3 times at the interval of 14 days with purified p53 protein that had been expressed in E.coli. The eggs laid by these immunized hens were then collected during the whole period of the experiment to prepare IgY from egg yolk by ammonium sulfate precipitation. Final purification and identification of the IgY were performed using SDS-PAGE, enzyme-linked immunosorbent assay and Western blotting. RESULT: All immunized hens developed specific antibodies to p53 protein in contrast to the control ones. The highest titers of the IgY occurred 4 weeks after the first immunization and reached 1:10(6), which remained stable for up to 3 months. CONCLUSION: IgY technology is less costly, non-invasive, fast, simple and highly efficient to generate polyclonal antibodies.