Rapid and simple purification of elastin-like polypeptides directly from whole cells and cell lysates by organic solvent extraction
| Autor: | David H. Thompson, Ross VerHeul, Craig Sweet |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cell lysates Biomedical Engineering Chemical Fractionation medicine.disease_cause Article law.invention 03 medical and health sciences chemistry.chemical_compound law Escherichia coli medicine General Materials Science Chromatography Aqueous solution Chemistry Extraction (chemistry) Recombinant Proteins Elastin Solvent 030104 developmental biology Alcohols Yield (chemistry) Solvents Recombinant DNA Elastin like polypeptides Peptides |
| Zdroj: | Biomaterials Science. 6:863-876 |
| ISSN: | 2047-4849 2047-4830 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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