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Mempro™ Protein Production in Mammalian Cells System

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Creative Biolabs has proprietary Mempro™ protein production in mammalian cells system generating high-yield eukaryotic membrane proteins from numerous mammalian cell types including human embryonic kidney cells (HEK-293), baby hamster kidney cells (BHK-21), monkey kidney fibroblast cells (COS-7) and Chinese hamster ovary cells (CHO). CHO cells are the abundantly used cell lines in membrane protein production due to their growth ability in suspension systems and bioreactors. While HEK-293 cells are also used for membrane protein production due to these cells are straightforward to grow and transfect in culture. Mempro™ protein production in mammalian cells system has the ability to perform correct protein folding and post-translational modification.

Mempro™ Protein Production in Mammalian Cells System Figure 1. Illustration of a typical process to perform recombinant membrane protein production in a mammalian cell line. (Pharmaceuticals, 2013)

Creative Biolabs has the following different approaches to obtain high-quality membrane protein in mammalian cells according to the specific needs for membrane protein expression:

Transient transfection system allows for membrane protein production within a few days after the expression plasmid has been constructed, while stable transfection system takes several months to develop membrane protein production. Transient transfection system can depend upon using either cationic compound to facilitate the uptake of plasmid DNA into the cell or recombinant non-replicative viruses that have evolved to efficiently enter mammalian cells. HEK293 cells are transiently transfected using liposomes, calcium phosphate or PEG as transfection reagents. CHO cells are commonly used to stably express large quantities of recombinant membrane proteins. This process involves transfection of dihydrofolate reductase (DHFR)-deficient CHO cells with the gene of interest and a DHFR selection cassette. The transfected cells are then screened in the presence of methotrexate to obtain stably transfected cell pools. The selection and expansion process takes 2-3 months.

The drawbacks of transient transfection and stable transfection systems in mammalian cells are expensive and difficult to scale-up. Thus, we can perform adenoviral vector-mediated membrane protein production in mammalian cells. Once the recombinant virus has been constructed, membrane protein productin is a reproducible, efficient process, even on a large scale.

Eukaryotic membrane protein produced from Mempro™ protein production in mammalian cells system has the best structural and functional features can satisfy the following applications, but not limited:

Creative Biolabs can provide other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.

References:
J. Andrell, et al. (2013). Overexpression of membrane proteins in mammalian cells for structural studies. Mol. Membr. Biol., 30(1): 52-63.
K. Lundstrom (2010). Expression of mammalian membrane proteins in mammalian cells using Semliki Forest virus vectors. Methods Mol. Biol., 601: 149-163.
T. Lai, et al. (2013). Advances in Mammalian cell line development technologies for recombinant protein production. Pharmaceuticals, 6(5): 579-603.




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