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利用高效膜亲和色谱法快速分析和微量纯化蛋白质

Fast assay and mini-purification of protein by high performance membrane affinity chromatography.

作者信息

Zhou D, Zou H, Ni J, Yang L, Jia L, Zhang Y

机构信息

National Chromatography Research and Analysis Center, Dalian Institute of Chemical Physics, the Chinese Academy of Sciences.

出版信息

Chin J Biotechnol. 1998;14(4):233-40.

Abstract

Several affinity ligands of protein A, human immunoglobulin G, iminodiacetate (IDA)-Cu2+, and cibacron blue F3GA were coupled to GMA-modified cellulose membrane matrices. We prepared the columns based on the membrane supports in high performance membrane affinity chromatography (HPMAC). Next we evaluated nonspecific adsorption of proteins on the membrane media. The rapid assay of proteins on the affinity columns based on membrane supports was achieved. It was observed that the affinity columns based on membrane supports provided good reproducibility, high separation efficiency, and low pressure drop of the column. The columns with the membrane media could also be used for purification of proteins.

摘要

将蛋白A、人免疫球蛋白G、亚氨基二乙酸(IDA)-Cu2+和汽巴克隆蓝F3GA的几种亲和配体偶联到甲基丙烯酸缩水甘油酯(GMA)修饰的纤维素膜基质上。我们基于这些膜载体制备了用于高效膜亲和色谱(HPMAC)的柱。接下来,我们评估了蛋白质在膜介质上的非特异性吸附。实现了基于膜载体的亲和柱上蛋白质的快速检测。观察到基于膜载体的亲和柱具有良好的重现性、高分离效率和低柱压降。带有膜介质的柱也可用于蛋白质的纯化。

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