Enhanced antibody production following intermediate addition based on flux analysis in mammalian cell continuous culture.
Generally, mammalian cells utilize glucose and glutamine as primary energy sources. To investigate the effect of energy sources on metabolic fluxes and antibody production, glucose- or glutamine-limited serum-free continuous culture of hybridoma 3A21 cells, which produce anti-ribonuclease A antibody, was carried out. The cell volume and dry cell weight were evaluated under various steady-state conditions. The specific consumption and production rates were evaluated on the basis of dry cell weight. On the basis of these results, the fluxes of the metabolic pathway were calculated. It was found that increasing the specific growth rate causes the specific ATP and antibody production rates to decrease. The fluxes between malate and pyruvate also decreased with the increase in specific growth rate. To increase the ATP production rate under steady-state conditions by the enhancement of fluxes between malate and pyruvate, the reduced metabolic fluxes were increased by an intermediate (pyruvate, malate, and citrate) addition. As a result, higher specific ATP and antibody production rates were achieved following the intermediate addition at a constant dilution rate.
Omasa
Takeshi
T
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan. omasa@bio.eng.osaka-u.ac.jp
Furuichi
Keisuke
K
Iemura
Tomoya
T
Katakura
Yoshio
Y
Kishimoto
Michimasa
M
Suga
Ken-Ichi
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D006825:Hybridomas / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008954:Models, Biological
D000911:Antibodies, Monoclonal; D016895:Culture Media, Serum-Free