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Simultaneous Targeting of Requiem & Alg-2 in Chinese Hamster Ovary Cells for Improved Recombinant Protein Production

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Abstract

Apoptosis is known to be the main cause of cell death in the bioreactor environment, leading to the loss of recombinant protein productivity. In a previous study, transcriptional profiling was used to identify and target four early apoptosis-signaling genes: FADD, FAIM, Alg-2, and Requiem. The resulting cell lines had increased viable cell numbers and extended culture viability, which translated to increased protein productivity. Combinatorial targeting of two genes simultaneously has previously been shown to be more effective than targeting one gene alone. In this study, we sought to determine if targeting Requiem and Alg-2 was more effective than targeting Requiem alone. We found that targeting Requiem and Alg-2 did not result in extended culture viability, but resulted in an increase in maximum viable cell numbers and cumulative IVCD under fed-batch conditions. This in turn led to an approximately 1.5-fold increase in recombinant protein productivity.

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Acknowledgments

This work was supported by the Biomedical Research Council of the Agency for Science, Technology and Research (A*STAR), Singapore.

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Authors and Affiliations

  1. Department of Chemical Engineering and Chemical Technology, Imperial College London, London, SW7 2AZ, UK

    Yiping Lim & Athanasios Mantalaris

  2. Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01 Centros, Singapore, 138668, Singapore

    Yiping Lim, Miranda G. S. Yap & Danny C. F. Wong

  3. Department of Chemical and Biomolecular Engineering, National University of Singapore, 10 Kent Ridge Crescent, Singapore, 119620, Singapore

    Miranda G. S. Yap

Authors
  1. Yiping Lim
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  2. Athanasios Mantalaris
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  3. Miranda G. S. Yap
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  4. Danny C. F. Wong
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Correspondence to Danny C. F. Wong.

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Lim, Y., Mantalaris, A., Yap, M.G.S. et al. Simultaneous Targeting of Requiem & Alg-2 in Chinese Hamster Ovary Cells for Improved Recombinant Protein Production. Mol Biotechnol 46, 301–307 (2010). https://doi.org/10.1007/s12033-010-9304-3

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  • DOI: https://doi.org/10.1007/s12033-010-9304-3

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