[利用Streamline SP从大肠杆菌中分离纯化抗-HBsAg Fab片段的吸附条件优化]
[Adsorption condition optimization for anti-HBsAg Fab fragment separation and purification from E. coli using Streamline SP].
作者信息
Huang Yu-xian, Luo Rong-cheng, Ding Xue-mei, Zheng Da-yong, Fang Yong-xin
机构信息
Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2006 Apr;26(4):409-13.
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.
目的
优化阳离子交换色谱介质Streamline SP的吸附条件，用于从大肠杆菌中分离纯化抗HBsAg Fab片段。
方法
采用试管法，在不同pH值和离子浓度的平衡缓冲溶液中，测试阳离子交换色谱介质Streamline SP对目标蛋白的吸附情况。然后使用1 ml Streamline SP预装柱和28 ml Streamline SP自装柱进行阳离子交换色谱，验证吸附效果。
结果
根据试管法实验结果，pH为4.4、离子浓度为100至600 mmol/L的上样缓冲液，可使阳离子交换色谱介质Streamline SP对目标蛋白达到最佳吸附效果，经1 ml SP预装柱和28 ml Streamline SP自装柱的阳离子交换色谱验证。
结论
试管法筛选出的阳离子交换色谱最佳条件，可用于从大肠杆菌中分离纯化抗HBsAg Fab片段。
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