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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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|
|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|
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