Membrane Protein Production in Escherichia coli: Overview and Protocols

Production of biological molecules is a challenge for the next decade in the field of medicinal chemistry. After heterologous production, the biological molecule must be active, well defined homogenous and the cost of its production should remain low. An interesting example is given by the relative success of therapeutic antibodies. Twenty monoclonal therapeutic antibodies are presently on the market (Oldham and Dillman 2008). All of them are produced with the hybridoma technology, which significantly increases the social cost of treating corresponding diseases and prevents the worldwide distribution of these drugs. Smaller-sized antibody peptides, named nanobodies, are being produced in bacteria to circumvent the cost of the hybridoma technology. Although Escherichia coli is probably the most versatile and the cheapest host for protein production, several obstacles remain: inclusion bodies formation, LPS contamination, incomplete synthesis, degradation by proteases, and the lack of post-translational modifications.