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

• DOI: 10.1110/ps.036939.108
• Publication Date: 2008-09-10T01:09:50Z
• Authors: Smita Raghava; Bipasha Barua; Pradeep K. Singh; Mili Das; Lalima Madan; Sanchari Bhattach...; Kanika Bajaj; B. Gopal; Raghavan Varadarajan; Munishwar N. Gupta
• Keywords: Inclusion bodies

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 technique three‐phase partitioning (TPP) obtain correctly refolded active from solubilized bodies. TPP was used for refolding 12 different overexpressed E. In each case, protein by gave either higher yield than earlier reported method or succeeded where efforts have failed. TPP‐refolded were characterized and compared conventionally purified terms their spectral characteristics and/or biological activity. The methodology is scaleable parallelizable does not require subsequent concentration steps. This approach may serve as a useful complement existing strategies diverse