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Expression of Biologically Active Human Antithrombin III in Chinese Hamster Ovary Cells

Bio/Technology volume 5, pages 720–725 (1987)Cite this article

Abstract

Dihydrofolate reductase (DHFR) deficient chinese hamster ovary (CHO) cells were cotransfected with a plasmid containing the human antithrombin III (AT III) cDNA preceded by the SV40 early promoter, and plasmids containing the mouse DHFR cDNA under the control of either the mouse mammary tumor virus long terminal repeat (MMTV–LTR) promoter or without any known eukaryotic promoter. Cell clones were selected that expressed the DHFR–gene despite the absence of a promoter in the latter case. The AT III cDNA was efficiently expressed in both cell types. After amplification with methotrexate (MTX), various cell lines secreted up to 22 μg AT III/106 cells/24 h. AT III was isolated from the medium of cultured cells and shown to be indistinguishable from human plasma derived AT III by immunological criteria and biological activity.

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Author notes

  1. Hermann Ragg: Hoechst AG, 6230 Frankfurt 80, Fed. Rep. of Germany.

Authors and Affiliations

  1. Research Laboratories of Behringwerke AG, P.O. Box 1140, 3550, Marburg, Fed. Rep. of Germany

    Gerd Zettlmeissl, Hermann Ragg & Hermann E. Karges

Authors
  1. Gerd Zettlmeissl
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  2. Hermann Ragg
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  3. Hermann E. Karges
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Zettlmeissl, G., Ragg, H. & Karges, H. Expression of Biologically Active Human Antithrombin III in Chinese Hamster Ovary Cells. Nat Biotechnol 5, 720–725 (1987). https://doi.org/10.1038/nbt0787-720

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  • DOI: https://doi.org/10.1038/nbt0787-720

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