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| 疏水层析分离正确折叠与错误折叠的复合干扰素 | |
| 引用本文： | 赵荣志, 刘永东, 王芳薇, 李京京, 夏新, 苏志国,.疏水层析分离正确折叠与错误折叠的复合干扰素[J].生物工程学报,2005,21(3):451-455. |
| 作者姓名： | 赵荣志 刘永东 王芳薇 李京京 夏新 苏志国 |
| 作者单位： | 1. 北京科技大学土木与环境工程学院,北京,100083;中国科学院过程工程研究所生化工程国家重点实验室,北京,100080 2. 中国科学院过程工程研究所生化工程国家重点实验室,北京,100080 3. 中国科学院过程工程研究所生化工程国家重点实验室,北京,100080;大连理工大学,大连,116024;大连医科大学附属二院,大连,116027 4. 中国环境监测总站,北京,100029 |
| 基金项目： | 国家自然科学基金资助项目 (No .2 0 13 60 2 0 )~~ |
| 摘 要： | 采用疏水层析纯化重组复合干扰素,成功去除了复性过程中产生的错误折叠体、聚集体及杂蛋白,并考察了配基类型、盐浓度、pH值和流速对疏水层析纯化效果的影响,结果表明采用ButylSepharose 4FastFlow、硫酸铵初始浓度0.8mol/L、缓冲液pH值8.3、线流速为90cm h时疏水层析纯化效果最佳,最终目标蛋白反相高效液相色谱检测纯度达到99.6% ,还原及非还原型SDS PAGE电泳均呈单一条带,其比活为2.3×109IU/mg,回收率为36.7%。 |
| 关 键 词： | 复合干扰素 纯化 疏水层析 |
| 文章编号： | 1000-3061(2005)03-0451-05 |
| 修稿时间： | 2004年11月12 |
Separation of Correctly Refolded and Mis-refolded Consensus Interferon by Hydrophobic Interaction Chromatography |
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| ZHAO Rong-Zhi,LIU Yong-dong,WANG Fang-Wei,LI Jing-jing,XIA Xin,SU Zhi-guo.Separation of Correctly Refolded and Mis-refolded Consensus Interferon by Hydrophobic Interaction Chromatography[J].Chinese Journal of Biotechnology,2005,21(3):451-455. | |
| Authors: | ZHAO Rong-Zhi LIU Yong-dong WANG Fang-Wei LI Jing-jing XIA Xin SU Zhi-guo |
| Institution: | Civil & Environmental Engineering School, University of Science and Technology of Beijing, Beijing 100083, China. |
| Abstract: | Hydrophobic interaction chromatography was used to separate correctly refolded and mis-refolded consensus interferon. The effects of ligand types, salt concentration, pH and flow rate were investigated. The best result could be obtained by using Butyl Sepharose 4 Fast Flow, 0.8 mol/L of ammonium sulfate, pH 8.3 and 90cm/h of linear flow rate. Reverse-phase HPLC analysis showed the purity of the pooled fraction was as high as 99.6%. The specific activity of purified consensus interferon was 2.3 x 10(9) IU/mg, The mass recovery of targeth protein was 36.7%. |
| Keywords: | consensus interferon purification hydrophobic interaction chromatography |
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