Solubilization and Refolding of Inclusion Body Proteins
Expression of heterologous proteins in E. coli often leads to the formation of protein aggregates known as inclusion bodies (IBs). Inclusion body aggregates pose a major hurdle in the recovery of bioactive proteins from E. coli. Usage of strong denaturing buffers for solubilization of bacterial IBs results in poor recovery of bioactive protein. Structure–function understanding of IBs in the last two decades have led to the development of several mild solubilization buffers, which improve the recovery of bioactive from IBs. Recently, combinatorial mild solubilization methods have paved the way for solubilization of wide range of inclusion bodies with appreciable refolding yield. Here, we describe a simple protocol for solubilization and refolding of an inclusion body protein with appreciable recovery.
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Solubilization and Refolding of Inclusion Body Proteins
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Protein recovery from inclusion bodies of Escherichia coli using mild solubilization process
Article Open access 25 March 2015
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Acknowledgments
This work is supported by the core grant of the National Institute of Immunology, received from the Department of Biotechnology, Government of India.