Immunoaffinity Purification of Specific Immunoglobulin from Egg Yolk
利用鸡蛋特异性抗体IgY的免疫亲和色谱

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.