重组人干细胞因子的纯化  

Purification of Recombinant Human Stem Cell Factor

作　　者：吴军[1] 巩新[1] 唱韶红[1] 赵志虎[1] 马清钧[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《中国生物化学与分子生物学报》2004年第6期821-826,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家"8 63"高技术重大专项基金资助 (No .Z18 0 3 3 0 )~~

摘　　要：干细胞因子 (SCF)是一种重要的造血生长因子 ,它在自体造血干细胞动员、肿瘤放疗化疗的辅助治疗、贫血和其它血液病的治疗上具有良好的应用前景 .为建立一种实用的rhSCF制备工艺 ,从表达rhSCF的工程菌分离包涵体 ,用尿素变性 ,低浓度尿素溶液稀释复性 .复性液中的rhSCF经离子交换层析吸附、浓缩 ,凝胶排阻层析分离去除共价二聚体和疏水层析精纯化 ,获得纯化的rhSCF .纯化的rhSCF纯度大于 95 % ,回收率为 4 7% ,可促进人红白血病细胞TF - 1的生长 ,比活性为 0 .9× 10 6U mg ,与英国NIBSC的rhSCF标准品相似 .上述结果表明 ,所建立的制备高纯度rhSCF工艺高效简便 。The stem cell factor(SCF) is important hematopoitic growth factor. The SCF may be useful in hematopoitic progenitor mobilization, reducing the duration of neutropenia and thrombocytopenia following high-dose chemotherapy or irradiation, rescuing anemia and other hematopathy. To establish a useful technology for rhSCF production, rhSCF inclusion body was separated from recombinant E. coli and denatured with urea solution. The denatured rhSCF was renatured by low concentrated urea solution and the rhSCF in the renatured solution was adsobted and concentrated with ion exchange chromatography. Finally, the rhSCF was purified with gel exclusion chromatography and hydrophobic chromatography for removing covalent dimer and delicate purification. The purity of the rhSCF was higher than 95%, the recovery rate was 47%. The purified rhSCF stimulated human erythroleukemia cell TF-1 proliferation and the specific activity was 0.9×10 6 U/mg, which was almost the same as the standard rhSCF from National Institute of Biological Standards and Control(NIBSC), United Kindom.The results showed that a high efficiency purification technology for high purity rhSCF was established and the specific activity of the purified rhSCF was high.

关 键 词：人干细胞因子 表达与纯化 变性与复性 

分 类 号：Q78[生物学—分子生物学]