摘要
目的:优化人血小板衍生生长因子BB亚型(PGDF-BB)包涵体复性方法与纯化条件,获得具有较好生物活性的重组PGDF-BB蛋白。方法:对PGDF-BB包涵体以梯度尿素进行变性,选择最佳包涵体变性浓度;比较不同复性条件下的复性率,稳定PGDF-BB包涵体复性方法;参照该蛋白的理化性质,选择适合PGDF-BB重组蛋白的纯化方法。结果:原核系统内实现了PGDF-BB的高表达;通过优化包涵体复性方法,重组蛋白的包涵体复性率可达40%以上;经过多个纯化方法相结合,PGDF-BB的纯度达到95%。结论:通过实验条件的优化,提高了PGDF-BB包涵体复性率,获得高纯度、高生物活性的重组PGDF-BB蛋白。
Objective： To obtain recombinant platelet-derived growthfactor BB（PGDF-BB） protein with high biological activity by optimizing inclusion body renaturation methods and purification conditions. Methods： The gradient urea concentration was designed for inclusion body denaturation, and the best method for purification of PGDFBB recombinant protein was chosen according to its physicochemical properties. Results： The renature ratio of recombinant PGDF-BB exceeded 40%, and purity of PGDF-BB reached 95%. Conclusion： The optimized methods adopted in this study improve PGDF-BB renaturation ratio evidently and increase its biological activity effectively.
作者
孙卫国
郑本献
熊志红
杨秉芬
刘艳华
张灵霞
SUN Wei-Guo;ZHENG Ben-Xian;XIONG Zhi-Hong;YANG Bing-Fen;LIU Yan-Hua;ZHANG Ling-Xia(Army Key Laboratory for Tuberculosis Prevention and Control, Beijing Key Laboratory of New Techniques for Tuberculosis Diagnosis and Treatment, Institute for Tuberculosis Research, 309 Hospital of PLA, Beijing 100091;No.61 Research Institute of Equipment Development, Academy of Military Sciences, Beijing 100141 China)
出处
《生物技术通讯》
CAS
2018年第3期382-386,共5页
Letters in Biotechnology
基金
国家传染病防治重大专项(2013ZX-10003-006)
