Industrial production of recombinant therapeutics in Escherichia coli and its recent advancements

Abstract

Nearly 30% of currently approved recombinant therapeutic proteins are produced in Escherichia coli. Due to its well-characterized genetics, rapid growth and high-yield production, E. coli has been a preferred choice and a workhorse for expression of non-glycosylated proteins in the biotech industry. There is a wealth of knowledge and comprehensive tools for E. coli systems, such as expression vectors, production strains, protein folding and fermentation technologies, that are well tailored for industrial applications. Advancement of the systems continues to meet the current industry needs, which are best illustrated by the recent drug approval of E. coli produced antibody fragments and Fc-fusion proteins by the FDA. Even more, recent progress in expression of complex proteins such as full-length aglycosylated antibodies, novel strain engineering, bacterial N-glycosylation and cell-free systems further suggests that complex proteins and humanized glycoproteins may be produced in E. coli in large quantities. This review summarizes the current technology used for commercial production of recombinant therapeutics in E. coli and recent advances that can potentially expand the use of this system toward more sophisticated protein therapeutics.

Modeling the microbial growth of two Escherichia coli strains in a multi-substrate environment

“…High density cultures of several strains of E. coli are currently used to produce recombinant proteins due to its high volumetric productivity; see e.g. Shiloach J. and Fass R. (2005) and Huang et al (2012). This, in practice, has transformed E. coli into a "factory" of recombinant proteins and many pharmaceutical products are produced in this way.…”

Transgenic Expression and Identification of Recombinant Human Proinsulin in Peanut

“…Early on, insulin was purified from the bovine or porcine pancreas. At present, E. coli and Saccharomyces cerevisiae are the most common hosts for the production of human insulin (Thim et al 1986;Nilsson et al 1996;Gellissen and Hollenberg 1997;Kjeldsen 2000;Porro et al 2005;Huang et al 2012;Ferrer-Miralles and Villaverde 2013;Baeshen et al 2014). The large-scale manufacture of therapeutic insulin for humans has benefited tremendously from genetic engineering (Arakawa et al 1998;Walsh 2005;Nykiforuk et al 2006;FerrerMiralles et al 2009;Boyhan and Daniell 2011;Qian et al 2011).…”

Cloning, soluble expression, and purification of the RNA polymerase II subunit RPB5 fromSaccharomyces cerevisiae

“…[13][14][15] The expression of this gene was increased using 3% ethanol (v/v). 16 Purified RPB5 can uncover concrete knowledge of the structure, interaction partners, and function of RPB5 in the mechanisms of gene expression operating at the molecular level.…”