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Refolding Techniques for Recovering Biologically Active Recombinant Proteins from Inclusion Bodies
Liberal Arts Education Center, Aso Campus, Tokai University, Minamiaso, Kumamoto 869-1404, Japan
Measurement Solution Research Center, National Institute of Advanced Industrial Science and Technology, Tosu, Saga 841-0052, Japan
Interdisciplinary Graduate School of Engineering Science, Kyushu University, Kasuga, Fukuoka 816-8580, Japan
* Author to whom correspondence should be addressed.
Received: 12 December 2013; in revised form: 23 January 2014 / Accepted: 10 February 2014 / Published: 20 February 2014
Abstract: Biologically active proteins are useful for studying the biological functions of genes and for the development of therapeutic drugs and biomaterials in a biotechnology industry. Overexpression of recombinant proteins in bacteria, such as Escherichia coli, often results in the formation of inclusion bodies, which are protein aggregates with non-native conformations. As inclusion bodies contain relatively pure and intact proteins, protein refolding is an important process to obtain active recombinant proteins from inclusion bodies. However, conventional refolding methods, such as dialysis and dilution, are time consuming and, often, recovered yields of active proteins are low, and a trial-and-error process is required to achieve success. Recently, several approaches have been reported to refold these aggregated proteins into an active form. The strategies largely aim at reducing protein aggregation during the refolding procedure. This review focuses on protein refolding techniques using chemical additives and laminar flow in microfluidic chips for the efficient recovery of active proteins from inclusion bodies.
Keywords: protein refolding; recombinant protein; inclusion body; denatured protein; chemical additive; microfluidic chip
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Yamaguchi, H.; Miyazaki, M. Refolding Techniques for Recovering Biologically Active Recombinant Proteins from Inclusion Bodies. Biomolecules 2014, 4, 235-251.
Yamaguchi H, Miyazaki M. Refolding Techniques for Recovering Biologically Active Recombinant Proteins from Inclusion Bodies. Biomolecules. 2014; 4(1):235-251.
Yamaguchi, Hiroshi; Miyazaki, Masaya. 2014. "Refolding Techniques for Recovering Biologically Active Recombinant Proteins from Inclusion Bodies." Biomolecules 4, no. 1: 235-251.