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| 溶液环境对百日咳疫苗分离纯化的影响 | |
| 引用本文： | 吴铁,闭静秀,黄永东,张焱,孙李靖,孙春宝,苏志国.溶液环境对百日咳疫苗分离纯化的影响[J].生物工程学报,2008,24(7):1279-1289. |
| 作者姓名： | 吴铁 闭静秀 黄永东 张焱 孙李靖 孙春宝 苏志国 |
| 作者单位： | 1. 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;中国科学院研究生院,北京,100049 3. 北京科技大学土木与环境工程学院,北京,100083 |
| 基金项目： | 国家自然科学基金项目(Nos. 20536050, 20576136 & 20636010)资助。 |
| 摘 要： | 针对百日咳疫苗在低盐条件下不稳定, 容易聚集而导致层析过程收率低、分离度低的难题, 实验中选择脲作为稳定剂来改善百日咳疫苗所处的溶液环境, 并采用离子交换层析和凝胶过滤层析进行百日咳疫苗的分离纯化, 通过ELISA抗原活性测定和还原性SDS-PAGE等方法研究了脲对百日咳疫苗分离纯化的影响。结果表明, 在流动相中加入 2 mol/L脲作为稳定剂, 能显著提高离子交换层析和凝胶过滤层析中的PT和FHA活性回收率、凝胶过滤层析的分离度、PT和FHA的纯度。这些结果对百日咳疫苗的分离纯化和层析工艺优化提供了重要的依据和参考。 |
| 关 键 词： | 溶液环境 脲 百日咳疫苗 百日咳毒素 丝状血凝素 |
| 收稿时间： | 26 October 2007 |
Effect of Solution Environment on the Purification of Pertussis Toxin |
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| Tie Wu,Jingxiu Bi,Yongdong Huang,Yan Zhang,Lijing Sun,Chunbao Sun and Zhiguo Su.Effect of Solution Environment on the Purification of Pertussis Toxin[J].Chinese Journal of Biotechnology,2008,24(7):1279-1289. | |
| Authors: | Tie Wu Jingxiu Bi Yongdong Huang Yan Zhang Lijing Sun Chunbao Sun and Zhiguo Su |
| Institution: | Civil & Environmental Engineering School, University of Science and Technology of Beijing, Beijing 100083, China;National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100080, China;National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100080, China; Graduate School of Chinese Academy of Sciences, Beijing 100049, China;National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100080, China; Graduate School of Chinese Academy of Sciences, Beijing 100049, China;National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100080, China; Graduate School of Chinese Academy of Sciences, Beijing 100049, China;Civil & Environmental Engineering School, University of Science and Technology of Beijing, Beijing 100083, China;National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100080, China |
| Abstract: | The low recovery of pertussis toxin (PT) and the low resolving efficiency of chromatography, due to the instability of PT in low salt condition, are the main challenges for its purification. We aplied 2 mol/L urea to prevent the aggregation and disassociation of PT during the purification by ion-exchange chromatography (IEC) and gel filtration chromatography (GFC). The effect of urea on the purification of PT was studied by ELISA assay and non-reduced SDS-PAGE. The activity recoveries of PT and filamentous hemagglutinin (FHA) in IEC and GFC, the resolution efficiency in GFC and the purities of PT and FHA were improved obviously by adding 2 mol/L urea in the mobile phase. The results highlight the potential application of urea in the acellular pertussis vaccine (APV) manufacture procedure. |
| Keywords: | solution environment urea pertussis vaccine pertussis toxin (PT) filamentous hemagglutinin (FHA) |
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