Who does not know that E. coli is one of the most used microorganisms to overexpress recombinant proteins?

This small Gram-negative Enterobacteriaceae has revolutionized the Molecular Biology and Biochemistry relegating to the past the large amount of animal and plant tissues processing to obtain small amounts of protein.

There are uncountable molecular tools and protocols to transform E. coli  and to obtain recombinant proteins from it, like a huge catalog of expression plasmids combining promoters, origin of replications, signal sequences (i.e to periplasmic space or to inclusion bodies), a great number of engineered strains and well known strategies to culture E. coli in large fermenters.

However, sometimes these approaches are dispersed and only available at the theoretical level, it is difficult to find a strong catalogue of updated solutions. The cloning and production, even being straightforward processes, are also minefields that could stop your developments, poor growth of the host, inclusion bodies (IB) if you want soluble overexpression, protein inactivity, low yields or even not obtaining any protein and many more…

Starting with the gene sequence of your protein of interest, 53Biologics could guide you with the selection and troubleshooting of the suitable expression vector or base strain and the best production and downstream strategy, to achieve acceptable yields obtaining your purified protein.

More than 98% of proteins in our hands have improved their production and/or recovery yields

The main limitation of E. coli is related with its inability to modify translated proteins and sometimes these post-translational modifications are needed for the stability or activity of your protein of interest.

However, E. coli is one of the faster alternatives, due to its growth rates and the availability of many different options to transform and screen, you could have a selected transformant starting with a DNA or just the aminoacid sequence of interest in around one month (feasibility study). After that you will have the option to explore the portfolio of molecular solutions in subsequent strain optimization programs.

During the last years, 53Biologics has developed and gathered an efficient molecular biology toolkit (IPFree of proprietary strains, promoters, signal peptides, etc…) to improve protein expression that is now available for its clients.

If definitively E. coli is not the best option, 53Biologics would help you to find the best alternative host.

The production process does not have much difficulty either, particularly considering the experience acumulated in 53Biologics with this kind of processes and here you could jump from flask scale to hundred liters reactors also in weeks.