Abstract
Cell expansion, gene transfer and protein production were all executed with a single serum-free, animal protein-free commercial medium designed for suspension-adapted Chinese hamster ovary cells (CHO DG44). This is a most important process to consider for clinical production of recombinant proteins. The transfection with polyethylenimine (PEI) was shown here to be scalable using both stirred-tank bioreactors of 3- and 150-l and novel agitated cultivation vessels (50 ml ventilated centrifuge tubes and 1-l square-shaped glass bottles) that lack any instrumentation. The transient transfections spanned a range of working volumes from 2 ml to 80 l. The maximum transient recombinant antibody yield was 22 mg/l, the highest ever reported for a multiliter transfection in CHO. The transiently expressed protein had the same extent of glycosylation as the same antibody produced from a stably transfected recombinant CHO cell line.
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Acknowledgements
The authors thank Rachel Flaction, Ilda Muller, and Elisabeth Derow for technical assistance and Dr. Maria De Jesus for cell line AMW. Financial support for this work was provided by the Swiss CTI Innovation Promotion Agency.
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Muller, N., Derouazi, M., Van Tilborgh, F. et al. Scalable transient gene expression in Chinese hamster ovary cells in instrumented and non-instrumented cultivation systems. Biotechnol Lett 29, 703–711 (2007). https://doi.org/10.1007/s10529-006-9298-x
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DOI: https://doi.org/10.1007/s10529-006-9298-x