脲梯度体积排阻色谱复性并同时纯化rhSCF  

Urea gradient SEC in renaturation with simultaneous purification of rhSCF

作　　者：王骊丽[1] 耿信笃[1] 

机构地区：[1]西北大学现代分离科学研究所,陕西省重点实验室陕西西安710069

出　　处：《第四军医大学学报》2005年第15期1349-1351,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(29675017);陕西省自然科学基金(2003C111)

摘　　要：目的:用脲梯度体积排阻色谱(SEC)复性并同时纯化在大肠杆菌中高表达的人干细胞因子.方法:采用脲梯度SEC,根据蛋白质与小分子物质分离过程的迁移速度不同的原理,在色谱过程中通过脲浓度的线性梯度洗脱方式脱除变性剂脲而达到蛋白复性并同时纯化.结果:用脲梯度SEC复性重组人干细胞因子(rhSCF)能够使复性蛋白的产率提高,色谱过程中洗脱长度、流速以及蛋白浓度的变化对蛋白复性效率都有不同的影响.结论:用脲梯度SEC复性rhSCF可以使复性产率提高,其质量回收率从35.2%提高到51.4%.AIM： To study the application of urea gradient size exclusion chromatography （SEC） in the renaturation with simultaneous purification of recombinant human stem cell factor （rhSCF） expressed in E. coli. METHODS： SEC was performed by the property of different migration speed of protein and small molecules in gel. The urea was elutedby linear elution, while the protein was eluted by urea gradient, thus resulting in the renaturation and purification of the denatured protein. RESULTS： The human stem cell factor was renatured and purified successfully using SEC. The efficiency was improved significantly. It was also found that the elution time, speed, as well as protein concentration had evident influence on the efficiency of protein renaturation. CONCLUSION： The human stem cell factor expressed in E. coli can be renatured and purified using SEC.With gradient elution, the efficiency of renaturation is improved and the recovery of the recombinant protein increases from 35.2% to 51.4%.

关 键 词：脲梯度 色谱法 重组人干细胞因子 大肠杆菌 

分 类 号：R378.21[医药卫生—病原生物学]