Method for Inclusion Bodies Production via E. coli Host System: rGCSF as Model Biotherapeutic Protein

Abstract

Escherichia coli is an industrial-relevant microbial host system, which is highly preferred for the large-scale production of recombinant biotherapeutics. Overexpression of these recombinant biotherapeutics in the E. coli system often results in the formation of insoluble protein aggregates termed as inclusion bodies (IBs). The yield and quality of IBs are affected by a spectrum of parameters like temperature, optical density, medium composition, induction time, and amount of inducer. Here, we present a protocol for the formation and processing of IBs for production of recombinant human granulocytes colony-stimulating factor (rGCSF).

Keywords: E. coli; Inclusion bodies; Induction phase; Recombinant protein; rGCSF.