( when thetop DO setpoint could not be...)Gram Quantities of MAb Produced with Simple Bioreactor in Serum-Free Perfusion Culture Replacing Ascitic Fluid Production

Gibcos Hybridoma Serum-Free Medium was used, supplemented with between 2 - 4 mM L-glutamine and 100 U/mL penicillin and 100 Ug/mL streptomycin. A separate 5% glucose solution was fed to the cultures when the perfusion rate reached 2 volumes/day to maintain the residual glucose concentration at > 1 g/L. The hybridoma cell line used produces an IgG₁ MAb. Perfusion at 1 volume/ day (based on the bed volume), was initiated 24 hours after the commencement of the run, with increases to 2 then 3 volumes/day also 24 hours apart. The perfusion rate was maintained at 3 volumes/day for the duration.

Results and Discussion
The MAb produced in these cultivations has been traditionally produced from ascitic fluid with an average yield after purification of 5 mg/mouse. Yields produced using a contract service in 10 & 250 L batch reactors produced ~35 mg/L before purification. Using the CelliGen Plus perfusion system, yields of between 4 - 5 times that of the batch fermentation were obtained, based on purified material from the perfusion system but unpurified material from the batch system ( see table ).

All AGEN staff found the CelliGen Plus bioreactor very user friendly, easily simplifying an otherwise complicated and laborious procedure. The bioreactors display and associated BioCommand software provided fermentation information at the touch of a fingertip and trend graphing of process controls could also be viewed via BioCommand software. These features made the data handling, interpreting and report writing much simpler, something much appreciated by all staff. For CelliGen Plus literature contact your local NBS sales representative or write us at bioinfo@nbsc.com.

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