Longterm Serum-Free Culture of Murine Hybridomas

We have established a sub-clone of the murine hybridoma C1E3, termed C1E3.10. This sub-clone was derived by adaptation to the serum-free/defined medium Excell 300 (1), followed by recloning for high antibody secretion. Production of monoclonal antibody in stationary flask cultures gave similar results in both serum containing and EXCELL 300 medium. Using this cell line we have monitored the production of monoclonal antibody in EXCELL 300 over an extended period of culture, greater than 75 days, in a small scale cell entrapment bioreactor, (Dynacell). The levels of glucose, lactic acid and glutamine in the reservoir of the bioreactor were monitored and analysed. Variations in concentration of lactic acid, in particular, had a profound effect on the rate of antibody production.