Single step protocol to purify recombinant proteins with low endotoxin contents
Single step protocol to purify recombinant proteins with low endotoxin contents
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DOI：
10.1016/j.pep.2005.09.027
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发表时间：
2006-04-01
影响因子：
1.6
通讯作者：
Donzeau, M
中科院分区：
文献类型：
--
作者：
Reichelt, P;Schwarz, C;Donzeau, M
内毒素是从大肠杆菌中纯化的重组蛋白的不需要的副产物。内毒素的固有毒性使其去除成为蛋白质在多种生物测定中的应用和安全肠外给药的重要步骤。本文描述的方法是一种一步方案，可有效去除大肠杆菌中过度表达的标记蛋白中紧密结合的内毒素。我们将亲和色谱与非离子洗涤剂洗涤步骤相结合，以去除最终产品中的大部分内毒素污染物。内毒素减少量低于 4 至 0.2 EU mg(-1)，蛋白质回收率接近 100%。由于这一新方案仅需要一个步骤即可同时纯化标记蛋白并消除内毒素，因此它在时间、精力和费用方面具有巨大的优势。 (c) 2005 Elsevier Inc. 保留所有权利。
Endotoxin is an unwanted by product of recombinant proteins purified from Escherichia coli. The inherent toxicity of endotoxins makes their removal an important step for the proteins' application in several biological assays and for safe parenteral administration. The method described in this paper is a one-step protocol which is effective at removing tightly bound endotoxin from over-expressed tagged proteins in E. coli. We combined affinity chromatography with a non-ionic detergent washing step, to remove most of the endotoxin contaminants from the end product. An endotoxin reduction of less than 4 to 0.2 EU mg(-1) was achieved with protein recovery close to a yield 100%. As this new protocol requires only one step to simultaneously purify tagged proteins and eliminate endotoxins, it represents a substantial advantage in time, effort, and expense. (c) 2005 Elsevier Inc. All rights reserved.