Membrane proteins such as receptors, ion channels, transporters and enzymes are key regulators of cellular function. Membrane proteins account for up to two thirds of known drugable targets, highlighting their critical pharmaceutical importance.
Despite importance, membrane proteins are notoriously difficult to prepare in pure, correctly folded form in sufficient quantity for drug discovery purposes. The challenging of recombinant membrane proteins production is because their expression can be toxic to cells or cause inclusion bodies, affects the protein yield, purification and research period.
Profacgen offers high-quality membrane protein production services for drug discovery. Our platform was designed specifically to address issues associated with the discovery of therapeutics for low abundance membrane proteins.
Our technology utilizes an E. coli expression system designed to enhance expression and to direct the recombinant protein to cytosolic aggregates known as inclusion bodies. This enables a yield of up to 10,000 folds higher than seen with conventional recombinant membrane protein expression technologies. Yields using this method are typically between 5-50 mg per liter of cell culture medium. The aggregated recombinant protein is then taken through a series of proprietary purification and refolding steps to generate functional, native protein.
Yeast cells expression is the most economical eukaryotic expression system for scale up production of proteins that require post-translational modifications. By selecting stable and durable strains in yeast, Profacgen offers a high-yield and high-quality membrane protein production. Consideration of all essential parameters for recombinant membrane protein production in yeast including promoters, optimal vector copies per cell, inducibility, signal peptides and cellular location, we can product most membrane protein of interested for our clients.
Expression systems utilizing mammalian cells for recombinant proteins are able to introduce proper protein folding, post-translational modifications, and product assembly, which are important for complete biological activity. A number of mammalian cell lines have been utilized with the most common being HEK 293 (Human embryonic kidney) and CHO (Chinese hamster ovary).
The baculovirus expression system utilizes recombinant baculoviruses for simple transduction of mammalian cells, allowing for production of milligram quantities of proteins. This technology has proven efficient for production of many eukaryotic membrane proteins since its quality controls system in the endoplasmic reticulum.
Cell-free expression is a promising tool for the fast and efficient production of membrane proteins, in particular for the target protein is toxic to host cell. Cell-free system is not restricted by origin, size or topology of the target, and the most important things is that it can produce the membrane proteins in detergent micelles or other mimetic system to maintain the integrity and functionality of the protein. Profacgen can provide cell free membrane expression services based on eukaryotes and prokaryotes system.
All the time, Profacgen is committing to address the unmet needs of drug discovery for human diseases treatment and cure that involves membrane proteins. We have cloned and expressed more than 200 different GPCRs and ion channels. The current rate of success in expression and refolding are more than 75% for any given new protein and has been steadily increasing.
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