Strategy for purifying maltose binding protein fusion proteins by affinity precipitation

Raghava, Smita and Aquil, Samina and Bhattacharyya, Sanchari and Varadarajan, Raghavan and Gupta, Munishwar N (2008) Strategy for purifying maltose binding protein fusion proteins by affinity precipitation. In: Journal of Chromatography A, 1194 (1). pp. 90-95.

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Abstract

The maltose binding protein (MBP) affinity tag has been extensively used for protein purification. A commercial grade cationic starch could precipitateMBPor anMBP-tagged protein quantitatively by simultaneous addition of 10% (w/v) polyethylene glycol (PEG) and 50mM calcium chloride. The precipitated MBP or MBP-tagged protein could be selectively dissociated by suspending the precipitate in 1M 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 $PEG/CaCl_2$ 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:	Journal Article
Publication:	Journal of Chromatography A
Publisher:	Elsevier
Additional Information:	Copyright of this article belongs to Elsevier.
Keywords:	Maltose binding protein;MBP affinity tag;MBP-fusion protein;Affinity precipitation;Protein purification;Cationic starch.
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited:	17 Jul 2008
Last Modified:	19 Sep 2010 04:47
URI:	http://eprints.iisc.ac.in/id/eprint/15074

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