4个品系罗非鱼血清免疫球蛋白的分离纯化及分子量测定  

Purification and molecular weight measurement of serum immunoglobulin of four tilapia species

作　　者：赵飞[1] 柯剑 姜兰[1] 谭爱萍[1] 吴雅丽[1,3] 薛慧娟[1,3] 

机构地区：[1]中国水产科学研究院珠江水产研究所,广东广州510380 [2]广东海大集团养殖技术中心,广东广州511400 [3]上海海洋大学水产与生命学院,上海200090

出　　处：《广东农业科学》2011年第21期138-140,共3页Guangdong Agricultural Sciences

基　　金：广东省教育部产学研结合项目(2010A09020029);中国水科院基本科研业务费(2011C001);广东省科技计划项目(2011B020307001)

摘　　要：利用A蛋白柱亲和层析法分离纯化了4个品系罗非鱼血清的免疫球蛋白(Ig)。优化的分离纯化条件为:进样量1.0mL血清,4℃结合3.5 h,流速1.3 mL/min。通过标准曲线法测定分离得到的免疫球蛋白最高浓度在1.87～2.95 mg/mL之间。绘制洗脱曲线,比较A蛋白柱亲和层析法分离纯化4个品系罗非鱼血清Ig的效果。SDS-PAGE电泳检测发现,4个品系罗非鱼血清Ig重链、轻链的分子量分别在80、30 ku左右。表明A蛋白柱亲和层析法可用于罗非鱼血清Ig的快速分离纯化。The purification of serum immunoglobulin of four tilapia species were carried out by protein A-sepharose affinity chromatography. Optimal conditions for purification were confirmed： 1.0 mL serum sample volume, binding reaction at 4℃ for 3.5 h, 1.3 mL/min flow velocity. The concentration of serum immunoglobulin was measured by standard curve method and was between 1.87 mg/mL and 2.95 mg/mL. The purification effect of serum immunoglobulin of four tilapia species was compared by elution curve. The results of SDS-PAGE showed that the molecular weights of heavy and light chain were about 80 ku and 30 ku, respectively. The results showed that protein A affinity chromatography could be a rapid and convenient method to purify serum immunoglobulin of Tilapia.

关 键 词：A蛋白柱 罗非鱼 免疫球蛋白 

分 类 号：Q965.125[生物学—昆虫学]