比较免疫磁珠法与A蛋白亲和层析法纯化兔抗血清中的IgG-《集美大学学报（自然科学版）》

文章信息/Info

Title:: Purification of IgG from Rabbit Sera by ImmunomagneticBeads and Protein A Affinity Chromatography

作者:: 郝华1; 鄢庆枇1; 2; 邹文政1; 2; 纪勇1; 2; (1.集美大学水产学院,福建厦门 361021;2.福建省高校水产科学技术与食品安全重点实验室,福建厦门 361021)

Author(s):: HAO Hua1; YAN Qing-pi1; 2; ZOU Wen-zheng1; 2; JI Yong1; 2; (1.Fisheries College,Jimei University,Xiamen 361021,China;2.Key Laboratory of Science and Technology forAquaculture and Food Safety(Jimei University),Fujian Province,Xiamen 361021,China)

Keywords:: immunomagnetic bead; protein A sepharose affinity chromatography; IgG purification

摘要:: 优化了免疫磁珠分离法纯化抗血清的实验条件，并与A蛋白亲和层析法比较纯化效果，以求得到一种简便、快速、高效的纯化IgG方法.结果表明：从5只新西兰兔获得210 mL试管凝集效价高达1∶5 120的兔抗鳗弧菌抗血清，磁珠与IgG结合10 min达到平衡，其最大结合量为0.227 mg/mL；免疫磁珠分离法与A蛋白亲和层析法得到的IgG纯度都很高，经过SDS-PAGE电泳都只得到25 ku和55 ku左右的两条带；免疫磁珠分离法得到的抗体ELISA效价高于A蛋白亲和层析法；免疫磁珠分离法反应所需时间（10 min）小于A蛋白亲和层析法所需的20 min，其得率（11.5 mg/mL）也高于A蛋白亲和层析法的10.25 mg/mL

Abstract:: Anti-sera were prepared from rabbit by injection of Vibrio anguillarum antigen,and then the IgG was purified from the anti-sera by the methods of immunomagnetic bead and protein A affinity chromatography.The results showed:210 mL anti-sera with an agglutination titer of 1∶5 120 were prepared from 5 New Zealand rabbits.The bonding of IgG and immunomagnetic beads reached equilibrium within 10 min,and the maximum bonding was 0.227 mg/mL.Purified IgG exhibited two bands in SDS-PAGE gel,the molecular weight of heavy chain and light chain of IgG were 55 ku and 25 ku,respectively.The IgG purified by immunomagnetic beads exhibited higher titer than that purified by protein A affinity chromatography.The process of immunomagnetic beads separation could be finished within 10 min which was shorter than that of protein A Sepharose affinity chromatography(20 min).The IgG recovery efficacy of immunomagnetic beads method(11.5 mg/mL)was higher than that of protein A sepharose affinity chromatography(10.25 mg/mL).The results indicated that immunomagnetic beads purified IgG conveniently,fast and efficiently,so they could be widely introduced to IgG purification