Applied Microbiology and Biotechnology

, Volume 83, Issue 1, pp 43–57

Expression of BHRF1 improves survival of murine hybridoma cultures in batch and continuous modes

  • Sandra Juanola
  • Joaquim Vives
  • Ernest Milián
  • Eva Prats
  • Jordi J. Cairó
  • Francesc Gòdia
Biotechnological Products and Process Engineering


Cell death by apoptosis limits growth and productivity in most animal cell cultures. It is therefore desirable to define genetic interventions to generate robust cell lines with superior performance in bioreactors, either by increasing specific productivity, life-span of the cultures or both. In this context, forced expression of BHRF1, an Epstein–Barr virus-encoded early protein with structural and functional homology with the anti-apoptotic protein Bcl-2, effectively protected hybridomas in culture and delayed cell death under conditions of glutamine starvation. In the present study, we explored the potential application of BHRF1 expression in hybridomas for long-term apoptosis protection under different biotechnological process designs (batch and continuous) and compared it to strategies based on Bcl-2 overexpression. Our results confirmed that long-term maintenance of the anti-apoptotic effect of BHRF1 can be obtained using bicistronic configurations conferring enhanced protection compared to Bcl-2, even in the absence of selective pressure. Such protective effect of BHRF1 is demonstrated both in batch and continuous culture. Moreover, a further analysis at high cell densities in semi-continuous perfusion cultures indicated that the mechanism of action of BHRF1 involves cell cycle arrest in G0–G1 state and this is translated in lower numbers of dead cells.


BHRF1 Hybridoma cells Apoptosis protection 


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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Sandra Juanola
    • 1
  • Joaquim Vives
    • 1
  • Ernest Milián
    • 1
  • Eva Prats
    • 2
  • Jordi J. Cairó
    • 1
  • Francesc Gòdia
    • 1
  1. 1.Departament d’Enginyeria Química, Escola Tècnica Superior d’Enginyeria (ETSE)Universitat Autònoma de BarcelonaBellaterraSpain
  2. 2.Departament de Biologia Molecular i CellularIBMB-CSICJordi Girona 18-26BarcelonaSpain

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