| 重组人Fab金属螯合层析法纯化条件的研究 |
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| 引用本文： | 朱迎春,王琰,刘群英,化冰,高荣凯. 重组人Fab金属螯合层析法纯化条件的研究[J]. 生物化学与生物物理进展, 1997, 24(2): 136-139 |
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| 作者姓名： | 朱迎春 王琰 刘群英 化冰 高荣凯 |
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| 作者单位： | 海军总医院中心实验科, 北京 100037;海军总医院中心实验科, 北京 100037;海军总医院中心实验科, 北京 100037;海军总医院中心实验科, 北京 100037;海军总医院中心实验科, 北京 100037 |
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| 基金项目： | 国家“863”计划资助项目. |
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| 摘 要： | 在重组人Fab(rh Fab)表达载体的羧基端插入六个组氨酸, 使其对金属螯合层析介质产生特异性吸附, 可用金属螯合亲和层析法进行分离纯化. 采用自制金属(铜、锌金属离子)螯合层析介质, 以pH和咪唑两种洗脱方法,对rh Fab段的纯化效果进行了探讨. 结果显示: 铜离子螯合层析介质比锌离子螯合层析介质对rh Fab的亲和能力更强; pH洗脱方法的重复性优于咪唑法; 金属铜离子螯合层析法对rh Fab进行一步纯化可得到纯度大于95%的rh Fab产品.
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| 关 键 词： | 组氨酸，乙肝病毒表面抗原（HBsAg），重组人Fab，金属螯合亲和层析 |
| 收稿时间： | 1996-03-12 |
| 修稿时间： | 1996-07-19 |
Investigation of Metal Chelate Chromatography in Purification of Recombinant Human Fab |
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| ZHU Yingchun,WANG Yan,LIU Qunying,HUA Bing and GAO Rongkai. Investigation of Metal Chelate Chromatography in Purification of Recombinant Human Fab[J]. Progress In Biochemistry and Biophysics, 1997, 24(2): 136-139 |
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| Authors: | ZHU Yingchun WANG Yan LIU Qunying HUA Bing GAO Rongkai |
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| Affiliation: | Navy General Hospital, Beijing 100037, China;Navy General Hospital, Beijing 100037, China;Navy General Hospital, Beijing 100037, China;Navy General Hospital, Beijing 100037, China;Navy General Hospital, Beijing 100037, China |
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| Abstract: | Recombinant human Fab(rh Fab) with a hexahistidine tail attacked to the carboxyl end of Fd could be easily purified by immobilized metal ion affinity chromatography (IMAC). IMAC with different immobilized metal ions(Zn2+ and Cu2+) and different elution strategies (pH and imidazole gradient) were compared. The results showed that Cu2+ were more effective than Zn2+ in retaining the His tagged Fab protein. pH gradient showed better consistency in Fab recovery than imidazole gradient. Using Cu2+ IMAC more than 95% pure rh Fab could be obtained by one step purification. |
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| Keywords: | histidine anti-HBsAg recombinant human Fab immobilized metal ion affinity chromatography |
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