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Scalable transient gene expression in Chinese hamster ovary cells in instrumented and non-instrumented cultivation systems

Muller, Natalie ; Derouazi, Madiha ; Van Tilborgh, Frédéric ; Wulhfard, Sarah ; Hacker, David ; Jordan, Martin ; Wurm, Florian

In: Biotechnology Letters, 2007, vol. 29, no. 5, p. 703-711

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    Titre
    • Scalable transient gene expression in Chinese hamster ovary cells in instrumented and non-instrumented cultivation systems
    Auteur
    • Muller, Natalie. École Polytechnique Fédérale de Lausanne (EPFL), Institute of Bioengineering, Laboratory of Cellular Biotechnology, 1015, Lausanne, Switzerland
    • Derouazi, Madiha. École Polytechnique Fédérale de Lausanne (EPFL), Institute of Bioengineering, Laboratory of Cellular Biotechnology, 1015, Lausanne, Switzerland
    • Van Tilborgh, Frédéric. École Polytechnique Fédérale de Lausanne (EPFL), Institute of Bioengineering, Laboratory of Cellular Biotechnology, 1015, Lausanne, Switzerland
    • Wulhfard, Sarah. École Polytechnique Fédérale de Lausanne (EPFL), Institute of Bioengineering, Laboratory of Cellular Biotechnology, 1015, Lausanne, Switzerland
    • Hacker, David. École Polytechnique Fédérale de Lausanne (EPFL), Institute of Bioengineering, Laboratory of Cellular Biotechnology, 1015, Lausanne, Switzerland
    • Jordan, Martin. École Polytechnique Fédérale de Lausanne (EPFL), Institute of Bioengineering, Laboratory of Cellular Biotechnology, 1015, Lausanne, Switzerland
    • Wurm, Florian. École Polytechnique Fédérale de Lausanne (EPFL), Institute of Bioengineering, Laboratory of Cellular Biotechnology, 1015, Lausanne, Switzerland
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Biotechnology Letters, 2007, vol. 29, no. 5, p. 703-711. Kluwer Academic Publishers
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:311642
    Summary
    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 (50ml ventilated centrifuge tubes and 1-l square-shaped glass bottles) that lack any instrumentation. The transient transfections spanned a range of working volumes from 2ml to 80l. The maximum transient recombinant antibody yield was 22mg/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