Hybridomas in a bioreactor cascade: modeling and determination of growth and death kinetics |
| |
Authors: | W A M Bakker T Schäfer H H Beeftink J Tramper C D De Gooijer |
| |
Institution: | (1) Department of Food Science, Food and Bioprocess Engineering Group, Wageningen Agricultural University, P.O. Box 8129, 6700 EV Wageningen, The Netherlands;(2) Department of the Science of Food of Animal Origin, Faculty of Veterinary Medicine, Utrecht University, P.O. Box 80175, 3508 TD Utrecht, The Netherlands |
| |
Abstract: | Hybridomas were cultured under steady-state conditions in a series of two continuous stirred-tank reactors (CSTRs), using a serum-free medium. The substrate not completely converted in the first CSTR, was transported with the cells to the second one and very low growth rates, high death rates, and lysis of viable cells were observed in this second CSTR. These conditions are hardly accessible in a single vessel, because such experiments would be extremely time-consuming and unstable due to a low viability. In contrast to what is often observed in literature, kinetic parameters could thus be derived without the neccessity for extrapolation to lower growth rates. Good agreement with literature averages for other hybridomas was found. Furthermore, showing that the reactor series is a valuable research tool for kinetic studies under extreme conditions, the possibility to observe cell death under stable and defined steady-state conditions offers interesting opportunities to investigate apoptosis and necrosis. Additionally, a model was developed that describes hybridoma growth and monoclonal antibody production in the bioreactor cascade on the basis of glutamine metabolism. Good agreement between the model and the experiments was found.Abbreviation MAb
Monoclonal antibody
Nomenclature
C
AConcentration of any (mol m-3) component A
D Dilution rate (s-1)
K
dDeath-rate constant (mol m-3)
K
lLysis-rate constant (mol m-3)
K
sMonod constant (mol m-3)
m Maintenance coefficient (mol cell-1 s-1)
q Specific consumption (mol cell-1 s-1) or production rate
t Time (s)
X Cell concentration (cell m-3)
Y Yield coefficient (cell mol-1)
Greek symbols
d
Specific death rate (s-1)
l
Specific lysis rate (s-1) of viable cells
net
Net specific growth (s-1) rate
true
True specific growth (s-1) rate |
| |
Keywords: | hybridomas serum-free medium monoclonal antibodies reactor series kinetics modeling |
本文献已被 PubMed SpringerLink 等数据库收录! |
|