Effects of oxygen/glucose/glutamine feeding on insect cell baculovirus protein expression: a study on epoxide hydrolase production.
The recombinant protein yields for batch cultures of the insect cell baculovirus expression system have been significantly enhanced by oxygen, glucose, and glutamine feeding. The improvement in both volumetric and specific yields was based on influencing the metabolism of infected cells. Oxygen was absolutely required for viral replication and high protein expression in infected cells. Increases of 200% in volumetric yield and 100% in specific yield of recombinant epoxide hydrolase were achieved by controlling the dissolved oxygen (DO) level to near 35% saturation. An additional 100% increase was achieved by glucose and glutamine feeding. Results indicated that the intracellular metabolite pool was not adequate for recombinant protein overproduction. Finally, the specific protein yield, based on initial infection cell density, in high cell density spinner flasks and bioreactors of spent media with glucose and glutamine feeding was equivalent to that freshly diluted cultures.
Wang
M Y
MY
Center for Agricultural Biotechnology, Maryland Biotechnology Institute, College Park.
Kwong
S
S
Bentley
W E
WE
D000818:Animals; D016367:Baculoviridae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D001709:Biotechnology / Q000379:methods; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D004851:Epoxide Hydrolases / Q000096:biosynthesis* / Q000235:genetics; D005285:Fermentation; D015870:Gene Expression / Q000235:genetics; D005947:Glucose / Q000494:pharmacology*; D005973:Glutamine / Q000494:pharmacology*; D009036:Moths / Q000166:cytology* / Q000187:drug effects / Q000382:microbiology*; D010100:Oxygen / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D003470:Culture Media; D011994:Recombinant Proteins; D005973:Glutamine; D004851:Epoxide Hydrolases; D005947:Glucose; D010100:Oxygen