Production of an anti-HBsAg mouse IgG-2bκ monoclonal antibody in hollow fiber bioreactors using different cell culture media and operation modes

Serum-and protein-free media IMDM/SSR, RPMI/SSR, PFHM, TurboDomaHP-I, and TurboDomaHP-II were evaluated for anti-HBsAg CB.Hep-1 Mab production in hollow fiber bioreactors. RPMI 1640 and IMDM, supplemented with FCS, were used as controls. Initial testing of media in 75 cm T flasks demonstrated that achievable cell densities (0.13-2.39 x 10 cells/mL) and IgG concentrations (5.2-18.7 mg/mL), were very much media-dependent. Performance with PFHM and TurboDomaHP-I were best, and comparable with IMDM/5% fetal calf serum (FCS) in terms of maximum IgG concentration, while the usage of RPMI 1640 or TurboDomaHP-II as base media was not favorable for this hybridoma. TurboDomaHP-I was selected for further studies in Cell-Pharm1000 bioreactor, due to its optimization posibilities. IMDM/SSR was excluded from these experiments due to its high cost. Mab concentration increases in Cell-Pharm1000 began after 10 days and reached maximum values between 16 and 31 days, in correspondence with the medium and operation mode used. The only disadvantage observed with TurboDomaHP-I was the long cell latency phase. The latter led to the addition of 1% of FCS and 500 μM ferric citrate to this medium for scaling up studies in the Cell-Pharm2500 bioreactor. This medium fortification led to a drop in latency of 1.6 and 1.4 times, relative to values seen for TurboDomaHP-I and IMDM/5% FCS in Cell-Pharm1000 and also allowed the reclonning of cells extracted from the bioreactor extracapillary space without adaptation to another culture medium formulation. Our results indicate that Cell-Pharm2500 bioreactor could produce about 14.4 grams of this antibody per month, harvesting 420 mL every 48 hours if the plateau phase could be maintained for 30 days.