Effects of oxygen on recombinant protein production by suspension cultures of Spodoptera frugiperda (Sf-9) insect cells.
Spodoptera frugiperda (Sf-9) insect cells have been grown in serum-free medium in 250-ml spinner flasks. The maximum cell density obtained in these cultures was dependent on the aeration rate of the culture. Similar yields of uninfected cells were obtained when cultures were stirred in spinner flasks at 80 rev min-1 and in a 4-1 stirred-tank bioreactor and the dissolved oxygen in the bioreactor was controlled at 20% of air saturation. Cells were infected with a recombinant baculovirus at different multiplicities of infection: the timing and maximum level of expression of the recombinant protein were dependent on the multiplicity of infection, the cell density at infection, and on the aeration rate of the culture. Oxygen-limited growth resulted in undetectable levels of recombinant protein (< 6 ng recombinant protein 10(-7) cells). Compared with the maximum yields observed in spinner flask cultures, higher levels of recombinant protein were produced when cells were grown and infected in the bioreactor. The level of dissolved oxygen in the bioreactor was controlled at 50% of air saturation.
Wellcome Diagnostics Ltd, Wellcome Research Laboratories, Beckenham, UK.
R I
RI
Scott
J H
JH
Blanchard
C H
CH
Ferguson
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001709:Biotechnology; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D016174:Hepacivirus / Q000235:genetics; D007313:Insecta; D007700:Kinetics; D010100:Oxygen / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis* / Q000235:genetics
D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; D010100:Oxygen