Recombinant beta-galactosidase production in serum-free medium by insect cells in a 14-L airlift bioreactor.
Spodoptera frugiperda (Sf9) insect cells were successfully cultured in serum-free medium in a 14-L airlift bioreactor. Cell densities as high as 1 x 10(7) cells/mL were achieved with specific growth rates of approximately 0.0286 h-1 (doubling time of 24 h). This system was also used to demonstrate the expression of a reported gene, beta-galactosidase (beta-gal), when cells were infected with a recombinant baculovirus. Approximately 0.33 mg of beta-gal/mL (i.e., 104,000 units/mL) of medium were obtained at the 14-L scale, while about 0.95 mg of beta-gal/mL (i.e., 285,000 units/mL) of medium were obtained in small-scale shaker flasks. The difference was attributed to a suboptimal infection in the large scale. Specific oxygen consumption rates decreased from 5.58 x 10(-17) mol O2/cell.s in early exponential growth to 3.13 x 10(-17) mol O2/cell.s at 3 days post-infection.
King
G A
GA
Department of Chemical Engineering, Queen's University, Kingston, Ontario, Canada.
Daugulis
A J
AJ
Faulkner
P
P
Goosen
M F
MF
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001709:Biotechnology; D002455:Cell Division; D002460:Cell Line; D003470:Culture Media; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism; D007313:Insecta; D007700:Kinetics; D010101:Oxygen Consumption; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D001616:beta-Galactosidase / Q000096:biosynthesis* / Q000235:genetics
D003470:Culture Media; D011994:Recombinant Proteins; D001616:beta-Galactosidase; D005947:Glucose