A New Protocol for Solubilization, Refolding and Purification of Recombinant Human Granulocyte Colony-stimulating Factor in Inclusion Bodies  

作　　者：Jia Hua LIU Chao Zhan WANG Xin Du GENG 

机构地区：[1]Institute of Modem Separation Science, Key Laboratory of Separation Science in Shaanxi Province, Northwest University, Xi＇an 710069

出　　处：《Chinese Chemical Letters》2006年第6期799-802,共4页中国化学快报（英文版）

基　　金：This work is supported by the National Natural Science Foundation of china （No. 20175016 and No. 20475042） ;the Foundation of Key Laboratory of Modem Separation Science in Shaanxi Province （No. 05JS61）.

摘　　要：Recombinant human granulocyte colony-stimulating factor （rhG-CSF） in inclusion bodies was solubilized by 8 mol/L urea solution and subsequently precipitated by acetone to improve its purity. After that, the precipitates were solubilized by sodium hydroxide solution containing 2 mol/L urea. Then the solubilized rhG-CSF was passed through a size exclusion chromatography for refolding and extensive purification, and further purified by a weak anion exchange chromatography. The purity and mass recovery of refolded rhG-CSF were 96.5% and 75.6%, respectively. The bioactivity was 8.4x10^7 IU/mg.Recombinant human granulocyte colony-stimulating factor （rhG-CSF） in inclusion bodies was solubilized by 8 mol/L urea solution and subsequently precipitated by acetone to improve its purity. After that, the precipitates were solubilized by sodium hydroxide solution containing 2 mol/L urea. Then the solubilized rhG-CSF was passed through a size exclusion chromatography for refolding and extensive purification, and further purified by a weak anion exchange chromatography. The purity and mass recovery of refolded rhG-CSF were 96.5% and 75.6%, respectively. The bioactivity was 8.4x10^7 IU/mg.

关 键 词：Recombinant human granulocyte colony-stimulating factor alkaline solution solubilization of inclusion bodies protein refolding. 

分 类 号：Q51[生物学—生物化学]