Role of environmental conditions on the expression levels, glycoform pattern and levels of sialyltransferase for hFSH produced by recombinant CHO cells.
A recombinant CHO cell line in which the expression of human follicle stimulating hormone (hFSH) was under the control of the beta actin promoter was maintained in steady state perfusion cultures on a protein free medium. The level of expression of the hFSH was controlled by varying the steady state level of dissolved oxygen (10-90% of air saturation) and of sodium butyrate (0-1.5mM). Under these conditions, the specific productivity of hFSH (qFSH) varied from 0.7 to 4.8 ng hFSH/10(6) cells/h. As the specific productivity of hFSH increased, there was a shift in the FSH isoforms to the lower pI fractions, corresponding to increased sialic acid content. As the specific productivity of hFSH increased, shifting the isoform distribution towards the lower pI isoforms, that the sialyltransferase enzymic activity also increased.
Chotigeat
W
W
Department of Biotechnology, University of New South Wales, Sydney, Australia.
Watanapokasin
Y
Y
Mahler
S
S
Gray
P P
PP
D000199:Actins / Q000235:genetics; D000818:Animals; D001709:Biotechnology / Q000295:instrumentation / Q000379:methods; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D016466:CHO Cells; D006224:Cricetinae; D046508:Culture Techniques / Q000379:methods*; D005640:Follicle Stimulating Hormone / Q000096:biosynthesis*; D015870:Gene Expression; D006031:Glycosylation; D006801:Humans; D007700:Kinetics; D010100:Oxygen / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012794:Sialic Acids / Q000378:metabolism; D012799:Sialyltransferases / Q000378:metabolism*; D013997:Time Factors
D000199:Actins; D002087:Butyrates; D011994:Recombinant Proteins; D012794:Sialic Acids; D020148:Butyric Acid; D005640:Follicle Stimulating Hormone; D012799:Sialyltransferases; D010100:Oxygen