利用鸡蛋特异性抗体IgY的免疫亲和色谱

〖摘要：以牛血清白蛋白（BSA）作为抗原，利用偶联上BSA的Sepharose-4B的亲和色谱材料从鸡蛋蛋黄中一步分离特异性抗BSA抗体。经SDS-PAGE、双向免疫扩散检验，所洗脱的样品为电泳纯的特异性抗体。反之，将所得抗体再偶联到POROSHY上可分离抗原。同时考察了母鸡免疫过程中特异性抗体随时间的变化趋势。-ZY/摘要〗〖WX/文献〗1Scouten.W.H.AffinityChromatography，bioselectiveadsorptiononinertmatrices.NewYork:Wi

Immunoaffinity Purification of Specific Immunoglobulin from Egg Yolk

Immunoaffinity column (Sepharose 4B) was made with bovine serum albumin (BSA) and used to isolate anti BSA antibody from egg laid by hens which were immunized with BSA. Polyclonal antibody was eluted under different conditions (pH 5.0 2.8) because of its different affinity against antigen. SDS PAGE and double immunodiffusion analysis confirmed that antibody isolated from egg yolk was electrophoretically pure and specific. According to the separation aim in this paper, the final elution buffer was 0.1 mol/L glycine HCl buffer, pH 2.8. The final antibody yield was higher than 90%. As a new development in chromatographic media, POROS has its maximum pressure limit of 21 MPa. It has been widely used because of its high performance, high flow and large capacity. The sugar residue of the antibody was then oxidized and coupled to the hydrazide activated POROS HY. Pure targeted protein (BSA) was obtained through the POROS HY column. The tendency of specific antibody production was investigated during the immunization period. The amount of specific antibody has increased obviously after boost immunization.