Test bacterial inclusion body for activity prior to start denaturing and refolding processes to obtain active eukaryotic proteins

Abstract

One of a major drawbacks correlated with expressing antibody fragments in bacterial cells is insolubility, which is often regarded as an obstacle in obtaining active molecules. Recombinant proteins aggregated as inclusion bodies within bacterial cells are thought to be unfolded or misfolded, and therefore inactive. So, denaturing and refolding strategies, which are laborious and sometime inefficient, are used to obtain correctly-folded active proteins. In the current study, we show that large quantities of correctly folded and completely active scFv molecules are there in bacterial inclusion bodies; they only need to be isolated from inclusion bodies.

Keywords: Inclusion body; Native 3D structure; Non-denaturing condition; Refolding; ScFv.