Elastin-like polypeptides as a purification tag for recombinant proteins.
Elastin-like polypeptides as a purification tag for recombinant proteins.
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DOI：
10.1002/0471140864.ps0611s61
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发表时间：
2010-08
影响因子：
--
通讯作者：
Chilkoti, Ashutosh
中科院分区：
文献类型：
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作者：
Hassouneh, Wafa;Christensen, Trine;Chilkoti, Ashutosh
该单元介绍了一种重组蛋白纯化方法，该方法采用弹性蛋白样多肽（ELP）作为纯化标签。当 ELP 被加热至其下临界溶解温度 (LCST) 以上时，会发生急剧且可逆的相变。当 ELP 与蛋白质融合时，它们保留了这种行为，从而提供了一种从细胞污染物中分离重组 ELP 融合蛋白的简单方法，通过我们称为反向转变循环 (ITC) 的方法，使溶液循环通过 ELP 融合蛋白的不溶相和可溶相。该方法不需要使用色谱法，因此具有成本效益，易于放大和多重分析。
This unit presents a recombinant protein purification method that employs an Elastin-like polypeptide (ELP) as a purification tag. ELPs undergo a sharp and reversible phase transition when heated above their lower critical solution temperature (LCST). ELPs retain this behavior when they are fused to a protein, and thereby provide a simple method to isolate a recombinant ELP fusion protein from cell contaminants by cycling the solution through the insoluble and soluble phase of the ELP fusion protein by a method that we have termed Inverse Transition Cycling (ITC). This method does not require the use of chromatography, so that it is cost-effective, easy to scale up and to multiplex.