HIGH CELL-DENSITY AND HIGH MONOCLONAL-ANTIBODY PRODUCTION THROUGH MEDIUM DESIGN AND RATIONAL CONTROL IN A BIOREACTOR

A simple feeding strategy was developed and successfully employed for nutritional control in a 2-L fed-batch culture of hybridoma cells. A p reviously developed stoichiometric model for animal cell growth was us ed to design a supplemental medium for feeding. Undialyzed feta I bovi ne serum and trace metals (Fe2+, SeO32-, Li+, Zn2+, and Cu2+) were fed to the cells periodically in addition to the automatic feeding of oth er nutrients in the supplemental medium. In this study, the maximum vi able cell density was increased from 6.3 x 10(6) to 1.7 x 10(7) cells/ mL, and the culture span was extended from 340 to 550 hours. The fina l monoclonal antibody titer achieved was 2400 mg/L. The specific produ ction rates for ammonia and lactate were further reduced from 0.0045 a nd 0.0048 in our previous fed-batch experiments to 0.0028 and 0.0036 m mol/10(9) cell h, respectively. Only 3.4% of the total glucose consump tion was converted into lactate, compared to 67% in a conventional bat ch culture. (C) 1996 John Wiley & Sons, Inc.