The effect of oxidative stress on the production of the recombinant protein, interferon gamma, produced by Chinese hamster ovary cells in stirred-batch culture.
The CHO320 cell line, engineered to produce human interferon gamma was investigated with regard to its susceptibility to oxidative stress. Batch cultures of the cells grown in a bench-top bioreactor exhibited no marked response to changes in oxygen concentration between 6% and 14% whereas cell growth and recombinant protein production were inhibited by increasing the oxygen to 20%. High concentrations of hydrogen peroxide (in excess of 200 microM) were required to inhibit growth of the CHO320 cells whereas concentrations of 50 microns and 100 microM had no effect on recombinant protein production. Buthionine sulphoximine (50 microM and 100 microM) completely depleted the cells of glutathione within 24 h; however, no quantitative effect on recombinant protein production was seen. It is concluded that the CHO320 cells are, possibly as a consequence of the long selection process they have undergone, very resistant to oxidative stress.
Dunster
C A
CA
Biology and Biochemistry Department, Brunel University of West London, Middlesex, UK.
Cheeseman
K H
KH
Maddix
S P
SP
D000818:Animals; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D016466:CHO Cells; D006224:Cricetinae; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007371:Interferon-gamma / Q000096:biosynthesis*; D018384:Oxidative Stress; D011994:Recombinant Proteins
D011994:Recombinant Proteins; D019328:Buthionine Sulfoximine; D007371:Interferon-gamma; D006861:Hydrogen Peroxide; D005978:Glutathione