Transgene mRNA Levels and Stability are Key Factors to Enhance Transient Gene Expression in CHO DG44 Cells

  • Sarah Wulhfard
  • Divor Kiseljak
  • Lucia Baldi
  • David L. Hacker
  • Florian M. Wurm
Conference paper
Part of the ESACT Proceedings book series (ESACT, volume 5)

Abstract

The aim of this work was to identify some of the limiting factors in transient gene expression (TGE) in CHO cells and to propose strategies to overcome them. Increasing the amount of plasmid DNA in the transfection did not increase recombinant protein yields, and it had a negative impact on transgene mRNA levels. Therefore, two other strategies aimed at increasing transgene mRNA levels were investigated. The first involved hypothermic treatment of transfected cells and the second the addition of valproic acid (VPA) after transfection. Both strategies resulted in recombinant antibody yields of 40–60 mg/L, whereas the untreated control transfections produced only 5–10 mg/L. In the treated cultures, the steady-state level of transgene mRNA was 3–5 times higher than in the untreated cultures and remained stable up to 6 days post-transfection. The two strategies proposed here are cost-effective and scalable making large-scale TGE in CHO cells a feasible alternative for rapid production of gram amounts of recombinant protein.

Keywords

Chinese Hamster Ovary Cell Chinese Hamster Ovary Sodium Butyrate Recombinant Protein Production Transient Gene Expression 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Sarah Wulhfard
    • 1
  • Divor Kiseljak
    • 2
  • Lucia Baldi
    • 2
  • David L. Hacker
    • 2
  • Florian M. Wurm
    • 1
    • 3
  1. 1.Laboratory of Cellular BiotechnologySchool of Life Sciences, École Polytechnique Fédérale de LausanneLausanneSwitzerland
  2. 2.Laboratory of Cellular BiotechnologySchool of Life Sciences, Ecole Polytechnique FÉdÉrale de LausanneLausanneSwitzerland
  3. 3.ExcellGene S.A.MontheySwitzerland

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