Microbial platform technology for recombinant antibody fragment production: A review

Abstract

Recombinant antibody fragments are being used for the last few years as an important therapeutic protein to cure various critical and life threatening human diseases. Several expression platforms now days employed for the production of these recombinant fragments, out of which bacterial system has emerged a promising host for higher expression. Since, a small antibody fragment unlike full antibody does not require human-like post-translational modification therefore it is potentially expressed in prokaryotic production system. Recently, small antibody fragments such as scFvs (single-chain variable fragments) and Fabs (antibody fragments) which does not require glycosylation are successfully produced in bacteria and have commercially launched for therapeutic use as these fragments shows better tissue penetration and less immunogenic to human body compared to full-size antibody. Recently developed Wacker's ESETEC secretion technology is an efficient technology for the expression and secretion of the antibody fragment (Fab) exceeded up to 4.0 g/L while scFv up to 3.5 g/L into the fermentation broth. The Pfenex system and pOP prokaryotic expression vector are another platform used for the considerably good amount of antibody fragment production successfully. In this review, we summarize the recent progress on various expression platforms and cloning approaches for the production of different forms of antibody fragments in E. coli.

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

“…Codon optimization increases the expression of recombinant protein by many folds (Rosano and Ceccarelli, 2014;Gupta S. K. et al, 2019;Rosano et al, 2019). A redox environment and foldases [e.g., disulfide isomerases (Dsb proteins) and peptidyl-prolyl isomerases (PPIase)] are necessary to form the correct disulfide bond in the periplasm (Gupta and Shukla, 2017b). The incorporation of appropriate signal sequences for protein expression in periplasm or in the extracellular space aids correct protein folding and also minimum proteolytic degradation (Gupta and Shukla, 2016).…”

“…These antibody fragments show better tissue penetration and are less immunogenic to the human body in comparison to the full antibody. Recently, ESETEC secretion technology (Wacker Biotech) has been developed to secrete recombinant products into the culture broth during fermentation and resulted in a high yield of Fab (exceeding 4.0 g/l) and of scFv (up to 3.5 g/l) (Gupta and Shukla, 2017b). In another study, it was shown that the optimization of antibody fragment production was accompanied by the alleviation of stress production in the periplasm of E. coli.…”

Sophisticated Cloning, Fermentation, and Purification Technologies for an Enhanced Therapeutic Protein Production: A Review

“…The bacterial expression system is also used for the production of antibody fragments and its derivatives (e.g., ScFv, Fab, etc.) for the production at commercial scale and therapeutic human use (Gupta and Shukla, 2016a). Despite various advantages, the major disadvantage of the E. coli system is lack of post-translational machinery which leads to cumbersome expression and purification development (Mamat et al, 2015).…”

“…The selection of an expression platform is determined by its capability to give high productivity with desired product quality (Li et al, 2010; Rita Costa et al, 2010; De Jesus and Wurm, 2011). The CHO cells are mostly used as a host for the production of recombinant proteins, including mAbs and fusion proteins (Jayapal et al, 2007; Kelley, 2009; Li et al, 2010; Rita Costa et al, 2010; De Jesus and Wurm, 2011; Gupta and Shukla, 2015, 2016a). Recombinant interferons and tissue-type plasminogen activator (tPA) were the first proteins produced by CHO cells (De Jesus and Wurm, 2011).…”

Triggering outer membrane leakiness of a novel E. coli strain for recombinant protein production

“…In the biopharmaceutical sector in 2018, it produced 22% of all therapeutic proteins with active licenses in the USA and the EU [4]. Although this marked a decrease in the share of microbials on the biotherapeutic market compared to the decades before, the emergence of biologics, like antibody fragments (Fab), single domain antibodies (sdAb) and other scaffolds, which can be expressed in prokaryotes, leads to a resurge of E. coli as a production host [5][6][7][8].…”