Wang Chaozhan, Wang Lili, Geng Xindu
Key Laboratory of Separation Science in Shaanxi Province, Institute of Modern Separation Science, Department of Chemistry, Northwest University, Xi' an 710069, China.
Se Pu. 2007 Jul;25(4):514-7. doi: 10.1016/s1872-2059(07)60020-0.
Recombinant human granulocyte colony-stimulating factor (rhG-CSF) in the form of inclusion bodies expressed in Escherichia coli (E. coli) was simultaneously refolded and purified using protein folding liquid chromatography (PFLC). Cu2+ -iminodiacetic acid (IDA) Sepharose was selected as the stationary phase for immobilized metal ion affinity chromatography. rhG-CSF was purified and the aggregates were diminished under a linear gradient elution of imidazole in the presence of a suitable concentration of urea. Using only one PFLC run, the refolded rhG-CSF had a specific bioactivity of 1.8 x 10(8) IU/mg and a purity of 97%, with the mass recovery of 32%.
以包涵体形式在大肠杆菌中表达的重组人粒细胞集落刺激因子（rhG-CSF），使用蛋白质折叠液相色谱（PFLC）同时进行复性和纯化。选择Cu2+ -亚氨基二乙酸（IDA）琼脂糖作为固定金属离子亲和色谱的固定相。在合适浓度尿素存在下，通过咪唑线性梯度洗脱对rhG-CSF进行纯化并减少聚集体。仅通过一次PFLC运行，复性后的rhG-CSF比活性为1.8×10(8) IU/mg，纯度为97%，质量回收率为32%。
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