阴离子交换层析法纯化gp130单克隆抗体B-S12  

Purification of monoclonal antibody B-S12 from mouse ascites by strong anion exchange liquid chromatography

作　　者：马泓冰[1] 徐颖[1] 夏瑜[1] 朱一蓓[1] 庄羽美[1] 郁健峰[1] 张学光[1] 

机构地区：[1]苏州大学医学生物技术研究所,苏州215007

出　　处：《中国免疫学杂志》2006年第8期750-753,共4页Chinese Journal of Immunology

摘　　要：目的:建立一种从小鼠腹水中获得高纯度、活性好、纯化过程易于放大的抗gp130单克隆抗体BS12的纯化方法。方法:经硫酸铵沉淀等预处理后的腹水样品,在阴离子交换层析柱上进行分离纯化,用MTT法检测纯化后抗体的生物学活性。结果:腹水样品经硫酸铵沉淀、PBS复溶,用pH7.0、20mmol/LTrisHCl缓冲溶液稀释10倍后上强阴离子交换层析柱,NaCl梯度洗脱,可获得纯度大于95%的BS12抗体,回收率达73%,在体外对XG2细胞有明显的促增殖作用。结论:建立了快速高效从小鼠腹水中纯化BS12的方法,为该抗体的进一步应用提供了必要的实验基础。Objective：To develop a rapid method for the purification of monoelonal B-S12 from mouse ascites with high purity, high recovery and high biological activity. Methods：The aseites was subject to precipitation with ammonium sulfate, then purified by anion exchange column chromatography. The antibody＇s biological activity was tested by MTr method. Results：The method for purifying monoclonal antibody B-S12 from mouse ascites by anion exchange chromatography was established. The purity and recovery of B- S12 were up to 95% and 73% respectively. The purified antibody could stimulate the proliferation of XG-2 ceils significantly in vitro. Conclusion：The method was characterized with high purity, excellent bioactivity and high recovery. It provided an important basis for its further application.

关 键 词：阴离子交换层析法 纯化 单克隆抗体 B-S12抗体 腹水 

分 类 号：R392.11[医药卫生—免疫学]