重组人载脂蛋白A-I_((米兰变体))的复性及纯化  

Renaturation and Purification of Recombinant Human Apo A-I_M

作　　者：汪利俊[1] 丁满生[1] 郭美锦[1] 储炬[1] 庄英萍[1] 张嗣良[1] 谭小钉 刘大涛 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237 [2]上海信谊药业药物研究所,上海200085

出　　处：《中国生物制品学杂志》2004年第2期81-83,共3页Chinese Journal of Biologicals

基　　金：国家高技术研究发展计划 (2 0 0 2AA2 170 2 1);国家重大科技专项(2 0 0 2AA2Z3 45 1)

摘　　要：目的　从大肠杆菌中获得载脂蛋白A I(米兰变体 ) 的包涵体 ,对包涵体进行复性、纯化 ,最终获得具有生物活性的载脂蛋白A I(米兰变体 ) 。方法　包涵体以尿素溶解后 ,以疏水层析法复性重组蛋白 ;以离子交换层析对其进一步纯化 ,并对所得的蛋白进行生物活性分析。结果　经复性纯化的蛋白纯度达 95 % ,体外生物活性检测显示其具有生物活性。结论　本法能快速有效地对目的蛋白进行复性、纯化 。Objective To express the inclusion body containing Apo A I M in E.coli ,then renaturalize and purify to obtain biologically active Apo A I M.Methods Dissolve inclusion body with urea and renaturalize the recombinant protein by hydrophobic chromatography,then further purify by ion exchange chromatography.The obtained protein was subject to bioassay.Results The purity of recombinant protein after renaturation and purification reached 95%.The protein was proved to be biologically active by in vitro bioassay.Conclusion The method was suitable for the rapid and effective renaturation and purification of goal protein,so it is hopeful to be applied in industrial scale production.

关 键 词：大肠杆菌 载脂蛋白 米兰变体 复性 纯化 包涵体 生物活性 

分 类 号：R977.6[医药卫生—药品] Q51[医药卫生—药学]