Wu Min, Chen Qing, Wang Ya-xian
Department of Epidemiology, First Military Medical University, Guangzhou 510515, China.
Di Yi Jun Yi Da Xue Xue Bao. 2003 May;23(5):491-3.
To isolate immunoglobulin (IgY) from egg yolk of p53 protein-immunized hens and to study its reactivity to the antigens.
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.
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.
IgY technology is less costly, non-invasive, fast, simple and highly efficient to generate polyclonal antibodies.
从经p53蛋白免疫的母鸡蛋黄中分离免疫球蛋白（IgY），并研究其与抗原的反应性。
用在大肠杆菌中表达的纯化p53蛋白，以14天的间隔对产蛋母鸡进行3次免疫。在整个实验期间收集这些免疫母鸡所产的蛋，通过硫酸铵沉淀从蛋黄中制备IgY。使用SDS-PAGE、酶联免疫吸附测定和蛋白质免疫印迹法对IgY进行最终纯化和鉴定。
与对照母鸡相比，所有免疫母鸡均产生了针对p53蛋白的特异性抗体。IgY的最高效价在首次免疫后4周出现，达到1:10(6)，并在长达3个月的时间内保持稳定。
IgY技术成本较低、非侵入性、快速、简单且高效地产生多克隆抗体。
