中国地鼠卵巢细胞培养上清中重组骨形态发生蛋白2的三步纯化法(英文)  

A three-step purification method for rhBMP-2 produced in Chinese hamster ovary cells

作　　者：李睿书[1] 王石泉[1] 谭向阳[2] 刘建宁[1] 

机构地区：[1]南京大学生物制药工程技术中心,苏州215123 [2]华东师范大学分子与实验医药研究所,上海200062

出　　处：《南京大学学报（自然科学版）》2010年第1期85-91,共7页Journal of Nanjing University（Natural Science）

基　　金：The Doctoral Program of Higher Education (20060284045);the Ministry of Education of People’s Republic of China (20060284042,20060284045)

摘　　要：骨形态发生蛋白2(BMP-2)是转化生长因子β(TGF-β)超家族的成员,在骨组织工程领域被广泛应用.如今,BMP-2的商业生产所受关注程度与日俱增.这里建立了一种方便且低成本的方法,将人重组骨形态发生蛋白2(rhBMP-2)从中国地鼠卵巢细胞系(CHO)培养上清中纯化出来.该方法由三步组成:肝素亲和层析,Q-Sepharose离子交换层析和SephacrylS-200凝胶过滤,得率约29%.从1L培养上清中能得到约0.26mg的纯度在95%以上的rhBMP-2成熟二聚体,并通过异位成骨实验组织学分析证实了其生物活性.因此,该方法在CHO培养上清中纯化rhBMP-2方面具有潜在的应用价值.Bone morphogenetic protein-2 （BMP-2）,a member of the transforming growth factor β superfamily,is widely used in bone tissue engineering. Commercial production of BMP-2 is attracting increasing attention at present time. We have developed a convenient and low-cost purification method of human recombinant BMP-2 （rhBMP-2） from conditioned medium of Chinese hamster ovary cell line （CHO）. The three-step procedure with a purification yield of 29% includes heparin affinity chromatography,Q-Sepharose ion exchange chromatography and Sephacryl S-200 gel filtration. About 0.26 mg of purified rhBMP-2 dimer with purity of more than 95% was obtained from 1 L of conditioned medium. It was shown to be biologically active by a histological analysis of ectopic bone formation. In conclusion,this method has the potential application for rhBMP-2 production from CHO.

关 键 词：重组人骨形态发生蛋白2 中国地鼠卵巢细胞系 蛋白纯化 

分 类 号：Q51[生物学—生物化学] Q813.11