Citation: Jia Hua LIU, Chao Zhan WANG, Xin Du GENG. A New Protocol for Solubilization, Refolding and Purification of Recombinant Human Granulocyte Colony-stimulating Factor in Inclusion Bodies[J]. Chinese Chemical Letters, ;2006, 17(6): 799-802. shu

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

Institute of Modern Separation Science, Key Laboratory of Separation Science in Shaanxi Province, Northwest University, Xi'an 710069

Received Date: 1 December 2005

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

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 8.4×10⁷ IU/mg.
- Recombinant human granulocyte colony-stimulating factor,
- alkaline solution,
- solubilization of inclusion bodies,
- protein refolding
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