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Influence of bcl-2 on antibody productivity in high cell density perfusion cultures of hybridoma

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Abstract

Apoptosis is an active, genetically determined death mechanism which can be induced by a wide range of physiological factors and by mild stress. It is the predominant form of cell death during the production of antibodies from murine hybridoma cell lines. A number of studies have now demonstrated that the suppression of this death pathway, by means of over-expression of survival genes such as bcl-2, results in improved cellular robustness and antibody productivity during batch culture. In the present study, the influence of bcl-2 expression on hybridoma productivity in two high density perfusion bioreactor systems was investigated. In the first system, a fixed-bed reactor, the DNA content in the spent medium was 25% higher in the control (TB/C3-pEF) culture than that found in the bcl-2 transfected (TB/C3-bcl2) cultures at all perfusion rates. This is indicative of a higher level of cell death in the control cell line. The average antibody concentration for the TB/C3-pEF cell line was 14.9 mg L-1 at perfusion rates of 2.6 and 5.2 d-1. However, for the TB/C3-bcl2 cell line it was 33 mg L-1 at dilution rates of 2 and 4 d-1. A substantial increase in antibody concentration was also found in the Integra Tecnomouse hollow fibre reactor. The antibody titre in the TB/C3-bcl2 cassette was nearly 100% higher than that in the TB/C3-pEF cassette during the cultivation period which lasted 6 weeks. Clearly, these results demonstrate the positive impact of bcl-2 over-expression on production of antibody in hybridoma perfusion cultures.

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Authors and Affiliations

  1. Bioprozeß- und Bioverfahrenstechnik, Technische Universität Hamburg-Harburg, Denickestraße 15, D-21071, Hamburg, Germany

    D. Fassnacht, S. Rössing & R. Pörtner

  2. Centre for Bioprocess Engineering, School of Chemical Engineering, University of Birmingham, Edgbaston, Birmingham, B15 2TT, U.K

    R. P. Singh & M. Al-Rubeai

Authors
  1. D. Fassnacht
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  2. S. Rössing
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  3. R. P. Singh
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  4. M. Al-Rubeai
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  5. R. Pörtner
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Fassnacht, D., Rössing, S., Singh, R.P. et al. Influence of bcl-2 on antibody productivity in high cell density perfusion cultures of hybridoma. Cytotechnology 30, 95–106 (1999). https://doi.org/10.1023/A:1008055702079

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  • DOI: https://doi.org/10.1023/A:1008055702079

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