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Production of Recombinant Proteins in Chinese Hamster Ovary Cells Using A Protein-Free Cell Culture Medium

Bio/Technology volume 13, pages 389–392 (1995)Cite this article

Abstract

The growth-factor prototrophk Chinese hamster ovary (CHO) SSF3 cell line was previously adapted for growth in serum-free media. Here we present a newly designed medium which allows these cells to grow in the absence of any exogenously added growth factors. To investigate the capacity of CHO SSF3 cells for the efficient production of recombinant proteins in protein-free media, expression plasmids containing either a human single chain urokinase-type plasminogen activator (uPA)-encoding cDNA or a humanized immunoglobulin G (IgG) kappa light chain cDNA were introduced by transfection. The tryptophan synthase (trpB) gene of Escherichia coli was used as a dominantly acting selection marker allowing the cells to survive in a medium containing indole in place of tryptophan. Some of the clones obtained exhibited a stable uPA expression over a period of several months under selective conditions and the yields were up to 74 mg of uPA/l in a bioreactor and the productivity was around 40 mg/day per 109 cells. The yields of IgG light chains were up to 118 mg/l and the productivity was in the order of 56 mg/day per 109 cells in a bioreactor. These results demonstrate the potential of CHO SSF3 cells for the efficient production of recombinant proteins under protein-free conditions.

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Author notes

  1. Jakob Reiser

    Present address: National Institutes of Health, Bethesda, MD, 20892

  2. Jakob Reiser: Corresponding author.

Authors and Affiliations

  1. Institute of Biotechnology, Swiss Federal Institute of Technology, ETH-Hönggerberg, CH-8093, Zürich, Switzerland

    Michael Zang, Helmut Trautmann, Christine Gandor, Ferruccio Messi, Armin Fiechter & Jakob Reiser

  2. Department of Biotechnology, Ciba-Geigy Ltd., CH-4002, Basel, Switzerland

    Michael Zang, Christine Gandor, Fred Asselbergs & Christian Leist

  3. Dr. F. Messi Cell Culture Technologies, CH-8152, Glattbrugg/Zürich, Switzerland

    Ferruccio Messi

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Zang, M., Trautmann, H., Gandor, C. et al. Production of Recombinant Proteins in Chinese Hamster Ovary Cells Using A Protein-Free Cell Culture Medium. Nat Biotechnol 13, 389–392 (1995). https://doi.org/10.1038/nbt0495-389

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