Abstract
CHO cells that produce human recombinant beta-interferon (β-IFN) have been grown under several different culture conditions in an attempt to increase the yields of the therapeutic glycoprotein whilst maintaining consistent glycosylation and minimizing intermolecular aggregation. Supplementation of cultures with either sodium butyrate or sodium chloride increases the productivity of β-IFN. However, sodium butyrate alters the glycosylation profile with an increase in the proportion of more highly branched complex Nlinked glycans. A shift in temperature during the culture period from 37°C to 30°C maintains consistent glycosylation of β-IFN for long culture periods with increased overall productivity and reduced aggregation compared to 37°C cultures. The use of Cytopore 1 microcarriers that entrap the cells also increases volumetric productivity of non-aggregated β-IFN while maintaining consistent glycosylation throughout the culture period.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Andersen, D.C., Bridges, T., Gawlitzek, M., and Hoy, C., 2000, Multiple cell culture factors can affect the glycosylation of Asn-184 in CHO-produced tissue-type plasminogen activator, Biotechnol. Bioeng. 70: 25–31.
Bigge, J.C., Patel, T.P., Bruce, J. A., Goulding, P.M. Charles, S.M. and Parekh, R.B., 1995, Nonselective and efficient fluorescent labeling of glycans using 2-amino benzamide and anthranilic acid, Anal. Biochem. 230: 229–238.
Butler, M., 2005, Animal cell cultures: recent achievements and perspective in the production of biopharmaceuticals, Appl. Microbiol. Biotechnol. DOI: 10.1007/s00253-005-1980-8 in press.
Conradt, H.S., Egge, H., Peter-Katalinic, J., Reiser, W., Siklosi, T., and Schaper, K., 1987, Structure of the carbohydrate moiety of human interferon-b secreted by a recombinant Chinese hamster ovary cell line, J. Biol. Chem. 30: 14600–14605.
Giovannoni, G., Munschauer, F.E. 3rd, and Deinsenhammer, F., 2002, Neutralising antibodies to interferon beta during the treatment of multiple sclerosis, J. Neural Neurosurg. Psychiatry 73: 465–469.
Guile, G.R., Rudd, P.M., Wing, D.R., Prime, S.B., and Dwek, R.A., 1996, A rapid highresolution high-performance liquid chromatographic method for separating glycan mixtures and analyzing oligosaccharides profiles, Anal. Biochem. 240: 210–226.
Kagawa, Y., Takasaki, S., Utsumi, J., Hosoi, K., Shimizu, H., Kochibe, N., and Kobata, A., 1988, Comparative study of the asparagine-linked sugar chains of natural human interferon-β1 produced by three different mammalian cells, J. Biol. Chem. 33: 17508–17515.
Lamotte, D., Buckberry, L., Monaco, L., Soria, M., Jenkins, N., Engasser, J., and Marc, A., 1999, Na-butyrate increases the production and α2–6 sialylation of recombinant interferon-γexpressed by α2–6-sialyltransferase engineered CHO cells, Cytotechnol. 29: 55–64.
Karpusas, M., Nolte, M., Benton, C.B., Meier, W., Lipscomb, W.N. and Goelz, S., 1997, The crystal structure of human interferonβ-at 2.2-å resolution, Proc. Natl. Acad. Sci. 94: 11813–11818.
Kim, N.S. and Lee, G.M., 2002, Response of recombinant Chinese hamster ovary cells to hyperosmotic pressure: effect of Bcl-2 overexpression, J. Biotechnol. 95: 237–248.
Oh, S.K.W., Vig, P., Chua, F., Teo, W.K. and Yap, M.G.S., 1993, Substantial overproduction of antibodies by applying osmotic pressure and sodium butyrate, Biotechnol. Bioeng. 42: 601–610.
Olejnik, A., Grajek, W., and Marecik,R., 2003, Effect of hyperosmolarity on recombinant protein productivity in baculovirus expression system, J. Biotechnol. 102: 291–300.
Rodriguez, J., Spearman, M., Huzel, N., and Butler, M., 2005, Enhanced production of monomeric interferon-by CHO cells through the control of culture conditions, Biotechnol. Prog. 21: 22–30.
Runkel, L., Meier, W., Pepinsky, R.B., Karpusas, M., Whitty, A., Kimball, K., Brickelmaier, M., Muldowney, C., Jones, W., and Goelz, S.E., 1998, Structural and functional differences between glycosylated and non-glycosylated forms of human interferon-beta (IFN-beta), Pharm. Res. 15: 641–649.
Spearman, M., Rodriguez, J., Huzel, N., and Butler, M., 2005, Production and glycosylation of recombinant-interferon in suspension and Cytopore microcarrier cultures of CHO cells, Biotechnol. Prog. 21: 31–39.
Sung, Y.H. and Lee, G.M., 2005, Enhanced human thrombopoietin production by sodium butyrate addition to serum-free suspension culture of Bcl-2-overexpressing CHO cells, Biotechnol. Prog. 21: 50–57.
Sung, Y.H., Song, Y.J., Lim, S.W., Chung, J.Y., and Lee, M.G., 2004, Effect of sodium butyrate on the production, heterogeneity and biological activity of human thrombopoietin by recombinant Chinese hamster ovary cells, J. Biotechnol. 112: 323–335.
Wurm, F. M., 2004, Production of recombinant protein therapeutics in cultivated mammalian cells, Nature Biotechnol. 22: 1393–1398.
Yang, M. and Butler, M., 2000, Effects of ammonia on CHO cell growth, erythropoietin production, and glycosylation, Biotechnol. Prog. 68: 370–380.
Yang, M. and Butler, M., 2002, Effects of ammonia and glucosamine on the heterogeneity of erythropoietin glycoforms, Biotechnol. Prog. 18: 129–138.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Springer
About this paper
Cite this paper
Spearman, M., Rodriguez, J., Huzel, N., Sunley, K., Butler, M. (2007). Effect of Culture Conditions on Glycosylation of Recombinant beta-Interferon in CHO Cells. In: Smith, R. (eds) Cell Technology for Cell Products., vol 3. Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-5476-1_11
Download citation
DOI: https://doi.org/10.1007/978-1-4020-5476-1_11
Publisher Name: Springer, Dordrecht
Print ISBN: 978-1-4020-5475-4
Online ISBN: 978-1-4020-5476-1
eBook Packages: Chemistry and Materials ScienceChemistry and Material Science (R0)