Protein expression and production in mammalian cells
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The strategy for protein expression in mammalian cells is the transfection with plasmid vectors or infection with viral vectors (inducible or constitutive) of cell culture (adherent or suspension). The platform will perform cell cultures ready for transfection or infection with the vector carrying the recombinant gene provided by the user. In a second step, the platform will propose a set of expression vectors (commercial or developed on site) to be screened together with cell type for the optimization of functional and soluble protein and protein complexes production.
The platform will have a collection of mammalian cell lines available for the user and optimized for adherent or suspension cell cultures. The efficiency of transfection or infection will be checked and standardized with the expression of fluorescent proteins (GFP). The available expression vectors are plasmids and viruses. Most of the vectors are adapted to the gateway system. Vectors for constitutive or inducible expression are available. To set up the optimal protein expression conditions (cell type, cell density, expression vector, time of culture, plasmid or virus amount, concentration of inducer…) small culture volumes (50 ml) will be used and the protein expression will be tested by affinity purification and SDS-PAGE. The optimal conditions will be scaled up for production.