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Issue 4, 2018
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Rapid and simple purification of elastin-like polypeptides directly from whole cells and cell lysates by organic solvent extraction

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Abstract

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 (Tt) 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 Tt. Since the underlying basis for the ELP Tt 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.

Graphical abstract: Rapid and simple purification of elastin-like polypeptides directly from whole cells and cell lysates by organic solvent extraction

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Publication details

The article was received on 31 Jan 2018, accepted on 20 Feb 2018 and first published on 28 Feb 2018


Article type: Paper
DOI: 10.1039/C8BM00124C
Citation: Biomater. Sci., 2018,6, 863-876
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    Rapid and simple purification of elastin-like polypeptides directly from whole cells and cell lysates by organic solvent extraction

    R. VerHeul, C. Sweet and D. H. Thompson, Biomater. Sci., 2018, 6, 863
    DOI: 10.1039/C8BM00124C

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