重组人干细胞因子中试工艺研究  

Pilot Purification Procedure of Recombinant Human Stem Cell Factor

作　　者：刘玉应[1] 邓小晨[1] 刘忠荣 吴洽庆 陈松森[3] 

机构地区：[1]四川大学生命科学学院,成都610064 [2]成都地奥制药集团有限公司,成都610041 [3]中国医学科学院基础医学研究所,北京100005

出　　处：《应用与环境生物学报》2008年第2期261-264,共4页Chinese Journal of Applied and Environmental Biology

基　　金：国家“863”计划重大专项(2003AA2Z3360)~~

摘　　要：人干细胞因子(Human stem cell factor,hSCF)是一种重要的造血细胞因子,能刺激造血细胞的生长、增生与分化.本文在重组hSCF工程菌的基础上,通过高密度发酵培养、包涵体分离纯化、变复性、SP-Sepharose FF、Source 30RPC与Q-Sepharose FF层析等技术分离纯化出具有生物学活性的重组人干细胞因子,建立了适合大规模生产重组人干细胞因子的分离纯化工艺.纯化的rhSCF纯度达98%以上,生物活性为1·0×106IU/mg,各项质量检测指标均符合质量控制标准.该纯化工艺简单易行,一批次能制备克级样品,且产品回收率高,重复性好.Human stem cell factor （hSCF） is a hematopoietic cytokine that stimulates the survival, proliferation and differentiation of hematopoietic cells. In this study, a purification procedure of hSCF, including high density fed-batch culture, isolation of inclusion bodies, denaturation and refolding, followed by SP-Sepharose FF, Source 30 RPC and Q-Sepharose FF chromatographies, was developed. The purity and specific activity of the purified recombinant hSCF （rhSCF） were above 98% and 1.0 × 10^6 IU/mg, respectively. The results revealed that the rhSCF produced by this procedure corresponded to the standards of quality control. The milligram grade of the functional （rhSCF） with high purity was obtained by this procedure. This procedure is easy for operation, and it can isolate and purify the rhSCF at large-scale with high recovery rate. Fig 6, Tab 1, Ref 15

关 键 词：重组人干细胞因子 包涵体 复性 分离与纯化 生物学活性 

分 类 号：Q78[生物学—分子生物学]