p53蛋白的原核表达及其鸡卵黄抗体的制备

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

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∶106,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。

Objective To isolate imm un oglobulin (IgY) from egg yolk of p53 protein-immunized hens and to study its rea ctivity to the antigens. Method Immunization of egg-laying hens was performed f or 3 times at the interval of 14 days with purified p53 protein that had been e xpressed in E.coli. The eggs laid by these immunized hens were then collected du ring the whole period of the experiment to prepare IgY from egg yolk by ammoniu m sulfate precipitation. Final purification and identification of the IgY we re performed using SDS-PAGE, enzyme-linked immunosorbent assay and Weste rn blotting. Result All immunized hens developed specific antibodies to p53 p rotein 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 le ss costly, non-invasive, fast, simple and highly efficient to generate polyclona l antibodies.