Raghava, Smita ; Aquil, Samina ; Bhattacharyya, Sanchari ; Varadarajan, Raghavan ; Gupta, Munishwar N. (2008) Strategy for purifying maltose binding protein fusion proteins by affinity precipitation Journal of Chromatography A, 1194 (1). pp. 90-95. ISSN 0021-9673
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Official URL: http://www.sciencedirect.com/science/article/pii/S...
Related URL: http://dx.doi.org/10.1016/j.chroma.2008.04.029
Abstract
The maltose binding protein (MBP) affinity tag has been extensively used for protein purification. A commercial grade cationic starch could precipitate MBP or an MBP-tagged protein quantitatively by simultaneous addition of 10% (w/v) polyethylene glycol (PEG) and 50 mM calcium chloride. The precipitated MBP or MBP-tagged protein could be selectively dissociated by suspending the precipitate in 1 M NaCl. In the case of a soluble MBP fusion with a fragment of human immunodeficiency virus protein gp120, 38% of the contaminating proteins could be removed by precipitation with PEG/CaCl2 and 100% of the fusion protein was recovered. In all cases, the purified proteins showed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the expected changes in fluorescence emission spectra upon binding to maltose.
| Item Type: | Article |
|---|---|
| Source: | Copyright of this article belongs to Elsevier Science. |
| Keywords: | Maltose Binding Protein; MBP Affinity Tag; MBP-fusion Protein; Affinity Precipitation; Protein Purification; Cationic Starch |
| ID Code: | 57260 |
| Deposited On: | 26 Aug 2011 04:20 |
| Last Modified: | 26 Aug 2011 04:20 |
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