Effect of lipid supplements on the production and glycosylation of recombinant interferon-γ expressed in CHO cells

The effects of lipids on the glycosylation of recombinant human interferon-γ expressed in a Chinese Hamster Ovary cell line were investigated in batch culture. Lipids form an essential part of the N-glycosylation pathway, and have been shown to improve cell viability. In control (serum-free) medium the proportion of fully-glycosylated interferon-γ deteriorated reproducibly with time in batch culture, but the lipoprotein supplement ExCyte was shown to minimise this trend. Partially substituting the bovine serum albumin content of the medium with a fatty-acid free preparation also improved interferon-γ glycosylation, possibly indicating that oxidised lipids carried on Cohn fraction V albumin may damage the glycosylation process.

[1]  A. Bull,et al.  Glucose‐limited chemostat culture of chinese hamster ovary cells producing recombinant human interferon‐γ , 1992, Biotechnology and bioengineering.

[2]  R B Freedman,et al.  High-resolution separation of recombinant human interferon-gamma glycoforms by micellar electrokinetic capillary chromatography. , 1994, Analytical biochemistry.

[3]  R. Parekh,et al.  Different culture methods lead to differences in glycosylation of a murine IgG monoclonal antibody. , 1992, The Biochemical journal.

[4]  J. Young,et al.  Effect of Culture Conditions on IgM Antibody Structure, Pharmacokinetics and Activity , 1993, Bio/Technology.

[5]  S. Grzesiek,et al.  1H, 13C, and 15N NMR backbone assignments and secondary structure of human interferon-gamma. , 1992, Biochemistry.

[6]  M. Butler.  Mammalian cell biotechnology. A practical approach. , 1991 .

[7]  S E Ealick,et al.  Three-dimensional structure of recombinant human interferon-gamma. , 1991, Science.

[8]  K. Gull,et al.  Recombinant human interferon-gamma. Differences in glycosylation and proteolytic processing lead to heterogeneity in batch culture. , 1990, The Biochemical journal.

[9]  S. Krag,et al.  Dolichol metabolism in Chinese hamster ovary cells. , 1992, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[10]  J. Sambrook,et al.  Cell-free synthesis of enzymically active tissue-type plasminogen activator. Protein folding determines the extent of N-linked glycosylation. , 1992, The Biochemical journal.

[11]  D A Cumming,et al.  Glycosylation of recombinant protein therapeutics: control and functional implications. , 1991, Glycobiology.

[12]  E. Bause.  Model studies on N-glycosylation of proteins. , 1984, Biochemical Society transactions.

[13]  N Jenkins,et al.  Glycosylation of recombinant proteins: problems and prospects. , 1994, Enzyme and microbial technology.

[14]  P. G. Strange,et al.  Chinese hamster ovary cell growth and interferon production kinetics in stirred batch culture , 1991, Applied Microbiology and Biotechnology.

[15]  F. Franek,et al.  Serum-free medium for hybridoma and parental myeloma cell cultivation. , 1986, Methods in enzymology.

[16]  M. Farrar,et al.  The molecular cell biology of interferon-gamma and its receptor. , 1993, Annual review of immunology.

[17]  A. Bull,et al.  The effect of the dilution rate on CHO cell physiology and recombinant interferon‐γ production in glucose‐limited chemostat culture , 1993, Biotechnology and bioengineering.

[18]  A. Rosenwald,et al.  Regulation of glycosylation. Three enzymes compete for a common pool of dolichyl phosphate in vivo. , 1990, The Journal of biological chemistry.

[19]  C. Goochee,et al.  Glycosidase Activities in Chinese Hamster Ovary Cell Lysate and Cell Culture Supernatant , 1993, Biotechnology progress.

[20]  R. Freedman,et al.  Changes in the Glycosylation of Interferon-γ during Culture , 1993 .

[21]  Ylva Gavel,et al.  Sequence differences between glycosylated and non-glycosylated Asn-X-Thr/Ser acceptor sites: implications for protein engineering , 1990, Protein engineering.

[22]  R. Jefferis,et al.  Glycosylation of human IgG subclass and mouse IgG2b heavy chains secreted by mouse J558L transfectoma cell lines as chimeric antibodies. , 1993, Human antibodies and hybridomas.

[23]  M. Lehrman.  Biosynthesis of N-acetylglucosamine-P-P-dolichol, the committed step of asparagine-linked oligosaccharide assembly. , 1991, Glycobiology.

[24]  W. Miller,et al.  Production of tPA in recombinant CHO cells under oxygen‐limited conditions , 1993, Biotechnology and bioengineering.

[25]  C. Goochee,et al.  The Oligosaccharides of Glycoproteins: Bioprocess Factors Affecting Oligosaccharide Structure and their Effect on Glycoprotein Properties , 1991, Bio/Technology.