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

南京大学学报(自然科学版) ›› 2010, Vol. 46 ›› Issue (1): 85–91.

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

李睿书 1 , 王石泉 1 , 谭向阳 2 , 刘建宁 1

出版日期:2015-03-27 发布日期:2015-03-27
作者简介: (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)
基金资助:
The Doctoral Program of Higher Education (20060284045) , the Ministry of Education of People ’ s Republic of China (20060284042 ,20060284045)

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

L i Rui 2 S hu 1 , Wang S hi 2 Quan 1 , Tan Xiang 2 Yang 2 , L iu J ian 2 N ing 1

Online:2015-03-27 Published:2015-03-27
About author: (1.Bio 2 Pharmaceutical Engineering Research Center , Nanjing University , Suzhou , 215123 , China ;
2. Institute of Molecular and Experimental Therapeutics , East China Normal University ,
Shanghai , 200062 , China)

摘要/Abstract

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 -(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 2 Sepharose 离子交换层析和 Sephacryl S 2 200 凝胶过滤 , 得率约 29 %. 从 1 L 培养上清中能得到约 0 1 26 mg 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 .

Abstract: Bone morphogenetic protein 2-2 (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 1 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. Key words : 　 rhBMP - 2 , Chinese hamster ovary cell line , protein purification

李睿书 1 , 王石泉 1 , 谭向阳 2 , 刘建宁 1
. 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
[J]. 南京大学学报(自然科学版), 2010, 46(1): 85–91.

L i Rui 2 S hu 1 , Wang S hi 2 Quan 1 , Tan Xiang 2 Yang 2 , L iu J ian 2 N ing 1 . A three - step purification method for rhBMP -2 produced in Chinese hamster ovary cells

[J]. Journal of Nanjing University(Natural Sciences), 2010, 46(1): 85–91.

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