盐析与疏水层析相结合快速分离提纯猪胰激肽释放酶  

Rapid Purification of Kallikrein by the Combination of Hydrophobic Interaction Chromatography and Salt Precipitation

作　　者：李冲峰[1] 王仁伟[1] 刘淑珍[1] 谭天伟[2] 苏志国[1] 

机构地区：[1]中国科学院过程工程研究所生化工程国家重点实验室,北京100080 [2]北京化工大学生命科学与技术学院,北京100029

出　　处：《过程工程学报》2005年第5期550-553,共4页The Chinese Journal of Process Engineering

基　　金：国家自然科学重点基金资助项目(编号:20136020)

摘　　要：将疏水层析技术用于从猪胰脏中分离纯化激肽释放酶,建立了一种简便、快速的分离提纯方法:将粗品溶解后经过硫酸铵沉淀处理,然后经过Butyl Sepharose FF疏水层析后得到目标蛋白,分析其纯度大于500U/mg,盐析和疏水两步纯化的收率大于85.0%.同时比较了Phenyl Sepharose FF,Octyl Sepharose FF和Butyl Sepharose FF三种疏水介质分离纯化胰K的效果.本实验工艺与传统工艺相比,具有操作简单、快速、回收率和纯化倍数高等优点,有望成为一种从动物组织中快速分离纯化药用蛋白质的有效技术平台.A process involving salt precipitation and hydrophobic interaction chromatography was developed to purify kallikrein from swine pancreas. The crude extraction was precipitated by 35% and 70% saturation of ammonium sulfate solution separately to eliminate impurity protein and concentrate crude sample. Different hydrophobic media of Phenyl Sepharose FF, Octyl Sepharose FF and Butyl Sepharose FF were evaluated and Butyl Sepharose FF was screened to apply to the further chromatography purification. About 85% of the final recovery rate of kallikrein and over 500 U/mg purity were obtained after these two steps purification. This rapid and simple process is easily scaled up for the proteins preparation from animal organs and tissues source.

关 键 词：疏水层析 纯化 猪胰激肽释放酶 

分 类 号：Q81[生物学—生物工程] TQ02[化学工程]