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采用蛋白A亲和层析法对健康美洲黑石斑鱼血清中的免疫球蛋白(IgM)进行分离纯化。实验结果表明,蛋白A亲和层析法一步纯化即可得到电泳纯IgM,其重链和轻链的分子量约74.1 ku和26.0 ku。用纯化的黑石斑鱼IgM免疫实验兔,获得效价高达1∶128 000的兔抗黑石斑鱼IgM血清。Western-blot和间接ELISA检测证明,本工作制备的兔抗美洲黑石斑鱼IgM血清具有较高的特异性,为后续的黑石斑鱼免疫学研究奠定了基础。
Immunoglobulin (IgM) in serum of healthy American black grouper was purified by protein A affinity chromatography. The experimental results show that electrophoresis pure IgM can be obtained by one-step purification of Protein A affinity chromatography, with the molecular weights of heavy chain and light chain of about 74.1 ku and 26.0 ku. Experimental rabbits were immunized with purified black grouper IgM to obtain rabbit anti-black grouper IgM sera with a titer of up to 1: 128,000. The results of Western-blot and indirect ELISA showed that the rabbit anti-American black grouper IgM serum had higher specificity and laid the foundation for the subsequent immunological research of Black grouper.