Effect of addition of proton carriers in culture medium on growth and secretion of hybridoma cell line OKT3.
Monoclonal antibodies (MAb) constitute the centre of all in-vitro diagnostic measures and almost all in-vivo therapeutic manoeuvres now. Production emphasis for these antibodies is having a current shift from animal-based large-scale culture to in-vitro bioreactor-based high-density culture. One of the major difficulties in high-density culture is end-metabolite accumulation in batch and fed-batch cultures in the forms of H+, NH4+ etc.. thereby reducing cellular growth and secretions. In the present study, effects of added proton carries--NAD and NADP--over and above the metabolic pools of the molecules, were examined on the cellular growth and secretion kinetics. Although NADP fortification showed a remarkable improvement in cellular growth (time dependent 200-300% improvements compared to controls) and size, cumulative MAb titre was better with NAD fortification. Combined additional loads of the proton carriers would be interesting to study in high density culture conditions.
Datta
D
D
Cellular Engineering Laboratory, School of Biomedical Engineering, Indian Institute of Technology, Mumbai-400 076.
Kundu
P K
PK
Dasgupta
S
S
Fulzele
K
K
Sewlikar
S
S
D000465:Algorithms; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000187:drug effects / Q000378:metabolism*; D008239:Lysine / Q000494:pharmacology; D051379:Mice; D016853:Muromonab-CD3 / Q000096:biosynthesis*; D009243:NAD / Q000494:pharmacology; D011522:Protons; D013056:Spectrophotometry, Ultraviolet; D013268:Stimulation, Chemical
D003470:Culture Media; D016853:Muromonab-CD3; D011522:Protons; D009243:NAD; D008239:Lysine