Elastin-like polypeptides (ELP) are a well-known class of proteins that are being increasingly utilized in a variety of biomedical applications, due to their beneficial physicochemical properties. A unifying feature of ELP is their demonstration of a sequence tunable inverse transition temperature (T_t) that enables purification using a simple, straightforward process called inverse transition cycling (ITC). Despite the utility of ITC, the process is inherently limited to ELP with an experimentally accessible T_t. Since the underlying basis for the ELP T_t is related to its high overall hydrophobicity, we anticipated that ELP would be excellent candidates for purification by organic extraction. We report the first method for rapidly purifying ELP directly from whole E. coli cells or clarified lysates using pure organic solvents and solvent mixtures, followed by aqueous back extraction. Our results show that small ELP and a large ELP-fusion protein can be isolated in high yield from whole cells or cell lysates with greater than 95% purity in less than 30 min and with very low levels of LPS and DNA contamination.

中文翻译：

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通过有机溶剂提取直接从整个细胞和细胞裂解物中快速，简单地纯化弹性蛋白样多肽†

弹性蛋白样多肽（ELP）是众所周知的一类蛋白质，由于其有益的理化性质，它们正越来越多地用于各种生物医学应用中。ELP的一个统一特征是它们演示了序列可调的逆转变温度（T _t），该温度可使用称为逆转变循环（ITC）的简单，直接的过程进行纯化。尽管使用了ITC，但该过程本质上仅限于具有实验上可访问的T _t的ELP 。由于ELP T _t的基础与ELP的整体疏水性高有关，我们预计ELP将是有机萃取提纯的极佳候选者。我们报告了使用纯有机溶剂和溶剂混合物直接从整个大肠杆菌细胞或澄清的裂解物中直接纯化ELP的第一种方法，然后进行水反萃取。我们的结果表明，可以在不到30分钟的时间内，从全细胞或细胞裂解物中高纯度地分离出小ELP和大ELP融合蛋白，纯度超过95％，并且LPS和DNA污染水平非常低。