Recombinant protein expression and purification for therapeutic development or for in vitro studies can sometimes be a real challenge. It requires huge investments in both time and cost in order to obtain high yields of pure and active recombinant proteins. My aim in writing this post, is to give you a simple guide to easily establish optimal conditions for recombinant protein expression and production in E. Coli, of problematic unstable proteins of interest.
Mass spectrometry is a technology which is widely used in most scientific disciplines that require accurate and precise measurement of elemental and molecular components. Its use in the pharmaceutical sector is often associated with the Drug Discovery and Development process. The primordial step to analyze a sample is to perform a sample treatment to enable its study. It really requires know-how to obtain good quality results. The following tips and tricks can help you to rapidly select a protein purification strategy and to anticipate some possible problems.
Expression systems – removing their contaminants
CHO, E. Coli and Pichia are useful expression systems for drug production. This can be for proteins, enzymes or antibody productions. Research and development for therapeutic perspectives requires a high level of quality controls. Such expression systems contain +1000 host cell proteins (HCPs). A major challenge in biological component production is to remove this HCP contamination. In order to monitor removal efficiency (during protein purification, as mentioned further on in this article), Canopy Biosciences have developed and optimised HCP detection kits based on the principle of the sandwich ELISA test as illustrated below.
Decidedly, producing and purifying a protein is a form of art.
Indeed, each recombinant protein (rec. protein) to be produced in vitro has specific characteristics and singularities that make its production and purification most definitely challenging. In this post, I’d like to share and review a few tips and basics for optimal design of experiment when producing functional recombinant proteins and working on protein purification.
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