Optimization of a method for refolding of bacterial recombinant proteins

The over-expression of recombinant proteins in large amount is important for production of therapeutic proteins and structural study. There are several systems for expression of recombinant proteins. One of the most relevant expression systems is Escherichia coli (E. coli). Although this organism has many advantages, most of recombinant proteins expressed in E. coli hosts form inclusion bodies. For gaining biological activities, these structures should be refolded. Many techniques have been developed for in vitro protein refolding.
In this study, a method was designed for inclusion body solubilization and protein refolding. IBs were solubilized in the solution containing 2M urea. This is a mild solubilization method without creating random coil structures in the protein.
Inclusion bodies undergo mild solubilization with maintain native-like secondary structures. Solubilized proteins were refolded on chromatography column by using native buffer conditions. The results showed the recombinant proteins were purified with high efficiency without aggregation.
The results suggest that this method is easy, efficient, cheap procedure and usable for obtaining refolded recombinant proteins. In addition, purified protein with the method can be used in diagnosis and/or treatment of diseases.