Abstract
Serum-free culture of Chinese hamster ovary (CHO) cells has become increasingly common as a way of obtaining high levels of expression of recombinant proteins while simplifying recovery and downstream processing of the product. However, serum-free media may still contain one or more of a variety of animal-derived components including albumin, fetuin, various hormones and other proteins. We have demonstrated that it is possible to eliminate animal-derived proteins from a CHO medium formulation. Plasma protein fractions like albumin and fetuin may be replaced by plant-derived hydrolysates, resulting in medium that is protein-free but still undefined (CHO III PFM). CD CHO Medium is a chemically defined formulation which contains no protein or hydrolysates of either plant or animal origin. Peak cell densities and recombinant protein expression in CD CHO cultures compared favorably to expression in other media, although the maximal cell density and the highest levels of expression were observed at later time points. We were able to successfully supplement the culture with sodium butyrate to increase expression levels at the expense of peak cell density, so for recombinant cell lines showing an inverse relationship between growth and expression of recombinant product, strategies which limit the peak cell density may be useful for increasing expression.
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References
Jayme, D.W., Epstein, D.A., Conrad, D.R. (1988) Fetal bovine serum alternatives, Nature 334:547–548.
Rosa, M.D. (1989) Serum substitutes: is there a solution? BioPharm 2:16–17.
Lambert, K.J. and Birch, J.R. (1985) Animal Cell Biotechnology, R.E. Spier and J.B. Griffiths, Eds. (Academic Press, London), pp. 85–122.
Hall, C.V., Jacob, P.E., Ringlod, G.M., Lee, F. (1983) Expression and regulation of Escherichia coli lacZ gene fusions in mammalian cells. J Mol Appl Gen 2(1): 101–109.
Miller, J.H. (1972) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Kruh, J. (1982) Effects of sodium butyrate, a new pharmacological agent, on cells in culture. Mol Cell Biochem 42:65–82.
Oh, S.K.W., Vig, P., Chua, F., Teo, W.K., Yap, M.G.S. (1993) Substantial overproduction of antibodies by applying osmotic pressure and sodium butyrate. Biotechnol Bioeng 42:601–610.
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© 1998 Springer Science+Business Media Dordrecht
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Gorfien, S.F., Dzimian, J.L., Tilkins, M.L., Godwin, G.P., Fike, R. (1998). Recombinant Protein Production by CHO Cells Cultured in a Chemically Defined Medium. In: Nagai, K., Wachi, M. (eds) Animal Cell Technology: Basic & Applied Aspects. Animal Cell Technology: Basic & Applied Aspects, vol 9. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-5161-0_42
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DOI: https://doi.org/10.1007/978-94-011-5161-0_42
Publisher Name: Springer, Dordrecht
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