| 蛋白折叠液相色谱法复性和纯化大肠杆菌表达的重组人粒细胞集落刺激因子 |
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| 引用本文： | 王超展,王骊丽,耿信笃.蛋白折叠液相色谱法复性和纯化大肠杆菌表达的重组人粒细胞集落刺激因子[J].色谱,2007,25(4):514-517. |
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| 作者姓名： | 王超展 王骊丽 耿信笃 |
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| 作者单位： | Key Laboratory of Separation Science in Shaanxi Province, Institute of Modern Separation Science, Department of Chemistry, Northwest University, Xi’an 710069, China |
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| 基金项目： | 国家自然科学基金;陕西省重点实验室基金 |
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| 摘 要： | 用蛋白折叠液相色谱法(PFLC)对大肠杆菌表达的包涵体形式的重组人粒细胞集落刺激因子(rhG-CSF)进行了复性并同时纯化。用Cu2+-亚氨基二乙酸（IDA） Sepharose作为固定化金属离子亲和色谱的固定相。在低浓度脲存在下,以咪唑为洗脱剂,采用线性梯度洗脱rhG-CSF。该法仅通过一步PFLC分离，减少了复性过程中rhG-CSF的聚集,复性后的rhG-CSF的比活性为1.8×108 IU/mg,纯度为97%,质量回收率为32%。
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| 关 键 词： | 纯化 大肠杆菌 蛋白折叠 固定化金属离子亲和色谱 重组人粒细胞集落刺激因子 |
| 文章编号： | 1000-8713（2007）04-0514-04 |
| 收稿时间： | 2007-2-8 |
| 修稿时间： | 2007-02-07 |
Refolding and Purification of Recombinant Human Granulocyte Colony-Stimulating Factor from Escherichia coli by Using Protein Folding Liquid Chromatography |
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| WANG Chaozhan,WANG Lili,GENG Xindu.Refolding and Purification of Recombinant Human Granulocyte Colony-Stimulating Factor from Escherichia coli by Using Protein Folding Liquid Chromatography[J].Chinese Journal of Chromatography,2007,25(4):514-517. |
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| Authors: | WANG Chaozhan WANG Lili GENG Xindu |
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| Institution: | Key Laboratory of Separation Science in Shaanxi Province, Institute of Modern Separation Science, Department of Chemistry, Northwest University, Xi’an 710069, China |
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| Abstract: | 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%. |
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| Keywords: | immobilized metal ion affinity chromatography(IMAC) protein refolding purification recombinant human granulocyte colony-stimulating factor(rhG-CSF) Escherichia coli |
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