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High performance CHO cell line development platform for enhanced production of recombinant proteins including difficult-to-express proteins
BMC Proceedings volume 7, Article number: P75 (2013)
In an effort to improve product yield of mammalian cell lines, we have previously demonstrated that our proprietary DNA elements, Selexis Genetic Elements (SGEs), increase the transcription of a given transgene, thus boosting the overall expression of a therapeutic protein drug in mammalian cells . However, there are additional cellular bottlenecks, notably in the molecular machineries of the secretory pathways. Most importantly, mammalian cells have some limitations in their intrinsic capacity to manage high level of protein synthesis as well as folding recombinant proteins. These bottlenecks often lead to increased cellular stress and, therefore, low production rates.
Material and Methods
Our specific approach involves CHO cell line engineering. We constructed CHO-M libraries based upon the CHO-M genome and transcriptome and using unique proprietary transposon vectors harboring SGE DNA elements to compensate for rate-limiting factors . Each CHO-Mplus library displays a diversity of auxiliary proteins involved in secretory pathway machineries and cellular metabolism. Collectively, the libraries address a broad range of expression issues.
Our results demonstrate that components of the secretory and processing pathways can be limiting, and that engineering of the metabolic pathway ('omic' profiling) improves the secretion efficiency of therapeutic proteins from CHO cells.
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Ley D, Harraghy N, Le Fourn V, Bire S, Girod PA, Regamey A, Rouleux-Bonnin F, Bigot Y, Mermod N: MAR Elements and Transposons for Improved Transgene Integration and Expression. PLoS One. 2013, 8: e62784-
Le Fourn V, Girod PA, Buceta M, Regamey A, Mermod N: CHO cell engineering to prevent polypeptide aggregation and improve therapeutic protein secretion. Metab Eng. 2013, Feb 1. pii: S1096-7176(13)00002-5. doi: 10.1016/j.ymben.2012.12.003. [Epub ahead of print], [http://www.ncbi.nlm.nih.gov/pubmed/23380542]
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Girod, P., Le Fourn, V., Calabrese, D. et al. High performance CHO cell line development platform for enhanced production of recombinant proteins including difficult-to-express proteins. BMC Proc 7, P75 (2013). https://doi.org/10.1186/1753-6561-7-S6-P75
- Secretory Pathway
- Therapeutic Protein
- Mammalian Cell Line
- Protein Drug
- Clonal Cell Line