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Location: Protein Purification - Laboratory research > All News > Strategies for the purification and on-column cleavage of glutathione S-transferase fusion target proteins.
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Cyril Dian, Said Eshaghi, Thomas Urbig, Sean McSweeney, Anna Heijbel, Gilles Salbert and Darcy Birse. Journal of Chromatography B, 1 (2002), vol. 769, 133-144. In this report, we describe a flexible, efficient and rapid protein purification strategy for the isolation and cleavage of glutathione-S-transferase (GST) fusion proteins. The purification and on-column cleavage strategy was developed to work for the purification of difficult proteins and for target proteins where efficient fusion-tag cleavage is essential for downstream processes, such as structural and functional studies. To test and demonstrate the flexibility of this method, seven diverse unrelated target proteins were assayed. A purification technique is described that can be applied to a wide range of both soluble and membrane inserted recombinant target proteins of different function, structure and chemical nature. This strategy is performed in a single chromatographic step applying an on-column cleavage method, yielding “native” proteins in the 200 g to 40 mg/l scale of 95-98% purity. |