Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments

Abstract

Antibodies and antibody-derived molecules are continuously developed as both therapeutic agents and key reagents for advanced diagnostic investigations. Their application in these fields has indeed greatly expanded the demand of these molecules and the need for their production in high yield and purity. While full-length antibodies require mammalian expression systems due to the occurrence of functionally and structurally important glycosylations, most antibody fragments and antibody-like molecules are non-glycosylated and can be more conveniently prepared in E. coli-based expression platforms. We propose here an updated survey of the most effective and appropriate methods of preparation of antibody fragments that exploit E. coli as an expression background and review the pros and cons of the different platforms available today. Around 250 references accompany and complete the review together with some lists of the most important new antibody-like molecules that are on the market or are being developed as new biotherapeutics or diagnostic agents.

Keywords: E. coli; Fab; antibody fragment; scFv.

Figure 1

Schematic representation of optional folding and misfolding pathways for periplasmic recombinant protein.

Figure 2

Three step procedure for obtaining purified recombinant antibody fragments following the expression in E. coli.