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利用金属螯合POROS灌注层析介质快速纯化组氨酸标签的谷胱甘肽S-转移酶融合蛋白。

Rapid purification of histidine-tagged glutathione S-transferase fusion protein by metal chelate POROS perfusion chromatography media.

作者信息

Schmidbauer S B, Strobel O K

机构信息

Boehringer Mannheim GmbH, Penzberg, Germany.

出版信息

Front Biosci. 1997 Feb 15;2:c6-8. doi: 10.2741/a165.

Abstract

The use of POROS MC media allowed high speed column preparation, sample loading, and elution. Under these conditions the proteins of interest can be recovered under mild conditions to maintain maximum biological activity. The speed of the process also assures minimum degradation or post-extraction modifications of the protein of interest from exposure to naturally occurring proteases and other enzymes. It is evident from the analysis by polyacrylamide gel electrophoresis of the purified fusion protein and starting material that a high degree of purification is possible in a single high-speed separation on POROS MC. The high flow rate capabilities of POROS media offer several other significant advantages over conventional media. A few of these include high speed and systematic method optimization, rapid column preparation and sample loading, and fast time-saving chromatographic separations.

摘要

使用POROS MC介质可实现高速柱制备、样品加载和洗脱。在这些条件下,目标蛋白质可在温和条件下回收,以保持最大生物活性。该过程的速度还确保了目标蛋白质因暴露于天然存在的蛋白酶和其他酶而导致的降解或提取后修饰降至最低。通过对纯化的融合蛋白和起始材料进行聚丙烯酰胺凝胶电泳分析可知,在POROS MC上进行单次高速分离就有可能实现高度纯化。与传统介质相比,POROS介质的高流速能力还具有其他几个显著优势。其中包括高速和系统的方法优化、快速的柱制备和样品加载,以及省时的快速色谱分离。

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