南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (04): 409-413.

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用Streamline SP从大肠杆菌中初步分离纯化anti-HBsAg Fab最适吸附条件的选择及验证

黄宇贤； 罗荣城； 丁雪梅； 郑大勇； 方永鑫；   

  1. 南方医科大学南方医院肿瘤中心； 南方医科大学南方医院肿瘤中心 广东 广州 510515； 广东 广州 510515；
  • 出版日期:2006-04-20 发布日期:2006-04-20
  • 基金资助:
    国家自然科学基金(39670668)~~

Adsorption condition optimization for anti-HBsAg Fab fragment separation and purification from E.coli using Streamline SP

HUANG Yu-xian, LUO Rong-cheng, DING Xue-mei, ZHENG Da-yong, FANG Yong-xin Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China   

  1. 南方医科大学南方医院肿瘤中心； 南方医科大学南方医院肿瘤中心 广东 广州 510515； 广东 广州 510515；
  • Online:2006-04-20 Published:2006-04-20

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摘要： 目的选择阳离子交换介质Streamline SP分离纯化anti-HBsAg Fab的最适吸附条件。方法试管法分别测定平衡缓冲液在不同pH和不同离子强度下,阳离子交换介质Streamline SP对预分离纯化蛋白的吸附效果。用1 ml SP预装柱及自装28 ml Streamline SP柱进行离子交换层析以验证吸附效果。结果 NaAc-HAc作为平衡缓冲液在pH4．4、离子强度100～600mmol／L可以使阳离子交换介质Streamline SP与蛋白吸附达到最佳效果。在该条件下用1 ml SP预装柱及自装28 ml SP柱进行离子交换层析,均验证了这一吸附效果。结论试管法选择的离子交换层析的最适吸附条件用于从大肠杆菌中初步分离纯化anti-HbsAg Fab切实可行。 

Abstract: Objective To optimize the adsorption condition of cation-exchange chromatographic media Streamline SP for separation and purification of anti-HBsAg Fab fragment from E.coli. Methods The adsorption of the target protein for separation and purification by the cation-exchange chromatographic media Streamline SP was tested using test tube method in balanced buffer solution with different pH values and ion concentrations. The adsorption effect was then verified by cation-exchange chromatography using 1-ml Streamline SP prepacked column and 28-ml Streamline SP self-assembly column. Results According to the experiment results of test tube method, the loading buffer with pH of 4.4 and ionic concentration of 100 to 600 mmol/L could achieve optimal target protein adsorption effect by cation-exchange chromatographic media Streamline SP, as verified by cation-exchange chromatography with 1-ml SP prepacked column and 28-ml Streamline SP self-assembly column. Conclusion The optimal condition of cation-exchange chromatography selected by test tube method can be applied for separation and purification of anti-HBsAg Fab fragment from E.coli. 

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