Recombinant protein purification by self-cleaving aggregation tag.

@article{citeulike:2217656, abstract = {A simple technique is presented for non-chromatographic purification of recombinant proteins expressed in Escherichia coli. This method is based on a reversibly precipitating, self-cleaving purification tag. The tag is made up of two components: an elastin-like polypeptide (ELP), which reversibly self-associates in high-salt buffers at temperatures above 30 degrees C; and an intein, which causes the ELP tag to self-cleave in response to a mild pH shift. Thus, a tripartite ELP-intein-target protein precursor can be purified by cycles of salt addition, heating and centrifugation. Once purified, intein-mediated self-cleavage, followed by precipitation of the cleaved ELP tag, allows easy and effective isolation of the pure, native target protein without the need for chromatographic separations. Recoveries of 50-100 mg of cleaved, native target protein per liter of shake-flask culture have been achieved for over a dozen proteins, typically in 8-24 h depending on specific process parameters.}, address = {Department of Chemical Engineering, A-217 Engineering Quadrangle, Princeton University, New Jersey 08544-5263, USA.}, author = {Wu, W. Y. and Mee, C. and Califano, F. and Banki, R. and Wood, D. W. }, citeulike-article-id = {2217656}, doi = {http://dx.doi.org/10.1038/nprot.2006.314}, issn = {1750-2799}, journal = {Nat Protoc}, number = {5}, pages = {2257--2262}, posted-at = {2008-01-11 07:39:07}, priority = {2}, title = {Recombinant protein purification by self-cleaving aggregation tag.}, url = {http://dx.doi.org/10.1038/nprot.2006.314}, volume = {1}, year = {2006} }

TY - JOUR ID - citeulike:2217656 L3 - citeulike-article-id:2217656 N2 - A simple technique is presented for non-chromatographic purification of recombinant proteins expressed in Escherichia coli. This method is based on a reversibly precipitating, self-cleaving purification tag. The tag is made up of two components: an elastin-like polypeptide (ELP), which reversibly self-associates in high-salt buffers at temperatures above 30 degrees C; and an intein, which causes the ELP tag to self-cleave in response to a mild pH shift. Thus, a tripartite ELP-intein-target protein precursor can be purified by cycles of salt addition, heating and centrifugation. Once purified, intein-mediated self-cleavage, followed by precipitation of the cleaved ELP tag, allows easy and effective isolation of the pure, native target protein without the need for chromatographic separations. Recoveries of 50-100 mg of cleaved, native target protein per liter of shake-flask culture have been achieved for over a dozen proteins, typically in 8-24 h depending on specific process parameters. IS - 5 JF - Nat Protoc SN - 1750-2799 AD - Department of Chemical Engineering, A-217 Engineering Quadrangle, Princeton University, New Jersey 08544-5263, USA. EP - 2262 TI - Recombinant protein purification by self-cleaving aggregation tag. VL - 1 SP - 2257 AU - Wu, WY AU - Mee, C AU - Califano, F AU - Banki, R AU - Wood, DW PY - 2006/// UR - http://dx.doi.org/10.1038/nprot.2006.314 ER -