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Refolding and simultaneous purification by three-phase partitioning of recombinant proteins from inclusion bodies

Raghava, Smita and Barua, Bipasha and Singh, Pradeep K and Das, Mili and Madan, Lalima and Bhattacharyya, Sanchari and Bajaj, Kanika and Gopal, B and Varadarajan, Raghavan and Gupta, Munishwar N (2008) Refolding and simultaneous purification by three-phase partitioning of recombinant proteins from inclusion bodies. In: Protein Science, 17 (11). pp. 1987-1997.

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Abstract

Many recombinant eukaryotic proteins tend to form insoluble aggregates called inclusion bodies, especially when expressed in Escherichia coli. We report the first application of the technique of three-phase partitioning (TPP) to obtain correctly refolded active proteins from solubilized inclusion bodies. TPP was used for refolding 12 different proteins overexpressed in E. coli. In each case, the protein refolded by TPP gave either higher refolding yield than the earlier reported method or succeeded where earlier efforts have failed. TPP-refolded proteins were characterized and compared to conventionally purified proteins in terms of their spectral characteristics and/or biological activity. The methodology is scaleable and parallelizable and does not require subsequent concentration steps. This approach may serve as a useful complement to existing refolding strategies of diverse proteins from inclusion bodies.

Item Type: Journal Article
Additional Information: Copyright of this article belongs to Cold Spring Harbor Laboratory Press.
Keywords: three-phase partitioning;protein refolding;recombinant ribonuclease A;CcdB mutants;human CD4;inclusion bodies.
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited: 06 Jul 2009 12:08
Last Modified: 06 Jul 2009 12:08
URI: http://eprints.iisc.ernet.in/id/eprint/16826

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