Identifying the differences in mechanisms of mycophenolic acid controlling fucose content of glycoproteins expressed in different CHO cell lines.
In the biopharmaceutical industry, glycosylation is a critical quality attribute that can modulate the efficacy of a therapeutic glycoprotein. Obtaining a consistent glycoform profile is desired because molecular function can be defined by its carbohydrate structures. Specifically, the fucose content of oligosaccharides in glycoproteins is one of the most important attributes that can significantly affect antibody-dependent cellular cytotoxicity (ADCC) activity. It is therefore important to understand the fucosylation pathway and be able to control fucosylation at the desired level to match predecessor materials in late stage and biosimilar programs. Several strategies were explored in this study and mycophenolic acid (MPA) was able to finely modulate the fucose content with the least undesired side effects. However, the response was significantly different between CHO cell lines of different lineages. Further experiments were then performed for a deeper understanding of the mechanism of fucosylation in different CHO cell lines. Results indicated that changes in the intracellular nucleotide involved in fucosylation pathway after MPA treatment are the main cause of the differences in fucosylation level response in different CHO cell lines. Differences in MPA metabolism in the various CHO cell lines directly resulted in different levels of afucosylation measured in antibodies produced by the CHO cell lines. Biotechnol. Bioeng. 2016;113: 2367-2376. © 2016 Wiley Periodicals, Inc.
Cell Culture Development, Biogen, 5000 Davis Drive, Research Triangle Park, North Carolina, 27709. an.zhang@biogen.com.
An
A
Zhang
Cell Culture Development, Biogen, 5000 Davis Drive, Research Triangle Park, North Carolina, 27709.
Valerie Liu
VL
Tsang
Cell Culture Development, Biogen, Cambridge, Massachusetts, 02142.
Lam R
LR
Markely
Cell Culture Development, Biogen, Cambridge, Massachusetts, 02142.
Lutfiye
L
Kurt
Cell Culture Development, Biogen, 5000 Davis Drive, Research Triangle Park, North Carolina, 27709.
Yao-Ming
YM
Huang
Cell Culture Development, Biogen, Cambridge, Massachusetts, 02142.
Shashi
S
Prajapati
Cell Culture Development, Biogen, Cambridge, Massachusetts, 02142.
Rashmi
R
Kshirsagar
D000818:Animals; D016466:CHO Cells / Q000145:classification* / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods; D003412:Cricetulus; D005643:Fucose / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D009173:Mycophenolic Acid / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology
D006023:Glycoproteins; D005643:Fucose; D009173:Mycophenolic Acid
Chinese hamster ovary (CHO); antibody-dependent cellular cytotoxicity (ADCC); fucosylation; inosine monophosphate dehydrogenase (IMPDH); mycophenolic acid