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Related effects of cell adaptation to serum-free conditions on murine EPO production and glycosylation by CHO cells

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Abstract

The necessity to perform serum-free cultures to produce recombinant glycoproteins generally requires an adaptation procedure of the cell line to new environmental conditions, which may therefore induce quantitative and qualitative effects on the product, particularly on its glycosylation. In previous studies, desialylation of EPO produced by CHO cells was shown to be dependent on the presence of serum in the medium. In this paper, to discriminate between the effects of the adaptation procedure to serum-free medium and the effects of the absence of serum on EPO production and glycosylation, adapted and non-adapted CHO cells were grown in serum-free and serum-containing media. The main kinetics of CHO cells were determined over batch processes as well as the glycosylation patterns of produced EPO by HPCE-LIF. A reversible decrease in EPO production was observed when cells were adapted to SFX-CHOTM medium, as the same cells partially recovered their production capacity when cultivated in serum-containing medium or in the enriched SFMTM serum-free medium. More interestingly, EPO desialylation that was not observed in both serum-free media was restored if the serum-independent cells were recultured in presence of serum. In the same way, while the serum-independent cells did not release a sialidase activity in both serum-free media, a significant activity was recovered when serum was added. In fact, the cell adaptation process to serum-free conditions did not specifically affect the sialidase release and the cellular mechanism of protein desialylation, which appeared to be mainly related to the presence of serum for both adapted and non-adapted cells.

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Acknowledgments

The F. Le Floch fellowship and the project were financed by the Agence Nationale pour la Recherche Technique and Aventis Pharma (Paris, France). The authors are grateful to Dr. Abderrahim Mahfoudi (Aventis Pharma, Paris) for donating the cDNA coding for the EPO cassette, Fabrice Blanchard (LSGC, Nancy) for his technical support, and Professor Jean Straczek and Pascal Perrin (University H. Poincaré, Nancy) for their expert scientific assistance in capillary electrophoresis analysis.

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  1. François LeFloch

    Present address: Cayla-InvivoGen, 5, rue Jean Rodier, B.P. 4437, 31405, Toulouse Cedex 4, France

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  1. Laboratoire des Sciences du Génie Chimique – UPR CNRS 6811, Institut National Polytechnique de Lorraine, ENSAIA, 2, avenue de la Forêt de Haye, B.P. 172, 54505, Vandœuvre-lès-Nancy Cedex, France

    François LeFloch, Bertrand Tessier, Sébastien Chenuet, Jean-Louis Goergen & Annie Marc

  2. Aventis Pharma, 13, quai Jules Guesde, 94400, Vitry sur Seine, France

    François LeFloch, Jean-Marc Guillaume & Pierre Cans

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  1. François LeFloch
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Correspondence to Annie Marc.

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LeFloch, F., Tessier, B., Chenuet, S. et al. Related effects of cell adaptation to serum-free conditions on murine EPO production and glycosylation by CHO cells. Cytotechnology 52, 39–53 (2006). https://doi.org/10.1007/s10616-006-9039-y

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