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Enhanced antibody production following intermediate addition based on flux analysis in mammalian cell continuous culture

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Abstract

Generally, mammalian cells utilize glucose and glutamine as primary energy sources. To investigate the effect of energy sources on metabolic fluxes and antibody production, glucose- or glutamine-limited serum-free continuous culture of hybridoma 3A21 cells, which produce anti-ribonuclease A antibody, was carried out. The cell volume and dry cell weight were evaluated under various steady-state conditions. The specific consumption and production rates were evaluated on the basis of dry cell weight. On the basis of these results, the fluxes of the metabolic pathway were calculated. It was found that increasing the specific growth rate causes the specific ATP and antibody production rates to decrease. The fluxes between malate and pyruvate also decreased with the increase in specific growth rate. To increase the ATP production rate under steady-state conditions by the enhancement of fluxes between malate and pyruvate, the reduced metabolic fluxes were increased by an intermediate (pyruvate, malate, and citrate) addition. As a result, higher specific ATP and antibody production rates were achieved following the intermediate addition at a constant dilution rate.

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Acknowledgments

Parts of this paper were presented at the seventh international conference on Computer Applications in Biotechnology (CAB7) in Osaka, Japan. This work is partially supported by grants from the New Energy and Industrial Technology Development Organization of Japan, and the Program for the Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation, and a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Takeshi Omasa, Keisuke Furuichi, Tomoya Iemura, Yoshio Katakura & Ken-ichi Suga

  2. Department of Chemistry and Materials Technology, Kyoto Institute of Technology, Goshokaido-cho, Matsugasaki, Sakyo-ku, Kyoto, 606-8585, Japan

    Michimasa Kishimoto

  3. Meiji Dairies Corporation, 540 Naruda, Odawara, Kanagawa, 250-0862, Japan

    Keisuke Furuichi

  4. Kirin Brewery Company, Tokyo, Japan

    Tomoya Iemura

Authors
  1. Takeshi Omasa
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  2. Keisuke Furuichi
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  3. Tomoya Iemura
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  4. Yoshio Katakura
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  5. Michimasa Kishimoto
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  6. Ken-ichi Suga
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Correspondence to Takeshi Omasa.

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Omasa, T., Furuichi, K., Iemura, T. et al. Enhanced antibody production following intermediate addition based on flux analysis in mammalian cell continuous culture. Bioprocess Biosyst Eng 33, 117–125 (2010). https://doi.org/10.1007/s00449-009-0351-8

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  • DOI: https://doi.org/10.1007/s00449-009-0351-8

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