Skip to main content
SpringerLink
Log in

Analysis and Manipulation of Recombinant Glycoproteins Manufactured in Mammalian Cell Culture

  • Chapter
Handbook of Industrial Cell Culture

  • 583 Accesses

Abstract

Mammalian cell culture has progressed in recent years from a lab-scale enterprise, mainly used for in vitro testing of chemical agents, to a major source of therapeutic and prophylactic agents. Industrial-scale cell culture had its origins in vaccine production from adherent fibroblasts in roller bottles; however, the dominant technology employed today uses suspension-adapted Chinese Hamster ovary (CHO) or hybridoma cells in large-scale (up to 12,000 L) tanks.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 229.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 299.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 299.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Kornfeld, R. and Kornfeld, S. (1985) Assembly of asparagine-linked oligosaccharides. Annu. Rev. Biochem. 54, 631–664.

    Article  CAS  Google Scholar 

  2. Fagioli, C. and Sitia, R. (2001) Glycoprotein quality control in the endoplasmic reticulum. Mannose trimming by endoplasmic reticulum mannosidase I times the proteasomal degradation of unassembled immunoglobulin subunits. J. Biol. Chem. 276, 12,885–12,892.

    Google Scholar 

  3. Bianucci, A. M. and Chiellini, F. (2000) A 3D model for the human hepatic asialoglycoprotein receptor (ASGP-R). J. Biomol. Struct. Dyn. 3,435–451.

    Article  Google Scholar 

  4. Jefferis, R., Lund, J., and Pound, J. D. (1998) IgG-Fc mediated effector functions: molecular definition of interaction sites for effector ligands and the role of glycosylation. Immunol. Rev. 163, 59–76.

    Article  CAS  Google Scholar 

  5. Jackson, P. (1996) The analysis of fluorophore-labeled carbohydrates by polyacrylamide gel electrophoresis. Mol. Biotechnol. 5, 101–123.

    Article  CAS  Google Scholar 

  6. Lee, K. B., Loganathan, D., Merchant, Z. M., and Linhardt, R. J. (1990) Carbohydrate analysis of glycoproteins. A review. Appl. Biochem. Biotechnol. 23, 53–80.

    Article  Google Scholar 

  7. Sutton, C. W., O’Neill, J. A., and Cottrell, J. S. (1994) Site-specific characterization of glycoprotein carbohydrates by exoglycosidase digestion and laser desorption mass spectrometry. Anal. Biochem. 218, 34–46.

    Article  CAS  Google Scholar 

  8. James, D. C., Goldman, M. H., Hoare, M., Jenkins, N., Oliver, R. W. A., Green, B. N., et al. (1996) Post-translational processing of recombinant human interferon-gamma in animal expression systems. Protein Sci. 5, 331–340.

    CAS  Google Scholar 

  9. Hooker, A. D., Green, N. H., Baines, A. J., Bull, A. T., Jenkins, N., Strange, P. G., et al. (1999) Constraints on the transport and glycosylation of recombinant IFN-gamma in Chinese hamster ovary and insect cells. Biotechnol. Bioeng. 63, 559–572.

    Article  CAS  Google Scholar 

  10. James, D. C., Freedman, R. B., Hoare, M., Ogonah, O. W., Rooney, B. C., Larionov, O. A., et al. (1995) N-glycosylation of recombinant human interferon-y produced in different animal expression systems. Bio-technology 13, 592–596.

    CAS  Google Scholar 

  11. Ogonah, O. W., Freedman, R. B., Jenkins, N., Patel, K., and Rooney, B. C. (1996) Isolation and characterization of an insect-cell line able to perform complex N-linked glycosylation on recombinant proteins. Bio-technology 14, 197–202.

    CAS  Google Scholar 

  12. Kemp, P. A., Jenkins, N., Clark, A. J., and Freedman, R. B. (1996) The glycosylation of human recombinant alpha-1-antitrypsin expressed in transgenic mice. Biochem. Soc. Trans. 24, 339.

    Google Scholar 

  13. Jenkins, N., Parekh, R. B., and James, D. C. (1996) Getting the glycosylation right—implications for the biotechnology industry. Nat. Biotechnol. 14, 975–981.

    Article  CAS  Google Scholar 

  14. Hayter, P. M., Curling, E. M. A., Baines, A. J., Jenkins, N., Salmon, I., Strange, P. G., et al. (1991) Chinese hamster ovary cell growth and interferon production kinetics in stirred batch culture. Applied Microbio. Biotechnol. 34, 559–564.

    Article  CAS  Google Scholar 

  15. Regoeczi, E., Kay, J. M., Chindemi, P. A., Zaimi, O., Suyama, K. L.(1991) Transferrin glycosylation in hypoxia. Biochem. Cell Biol. 69, 239–244.

    Article  CAS  Google Scholar 

  16. Yang, M. and Butler, M. (2000). Effect of ammonia on the glycosylation of human recombinant erythropoietin in culture. Biotechnol. Prog. 16, 751–759.

    Article  CAS  Google Scholar 

  17. Borys, M. C., Linzer, D. I., and Papoutsakis, E. T. (1993) Culture pH affects expression rates and glycosylation of recombinant mouse placental lactogen proteins by Chinese hamster ovary (CHO) cells. Bio-technology 6, 720–724.

    Google Scholar 

  18. Grabenhorst, E., Schlenke, P., Pohl, S., Nimtz, M., and Conradt, H. S. (1999) Genetic engineering of recombinant glycoproteins and the glycosylation pathway in mammalian host cells. Glycoconj. J. 16, 81–97.

    Article  CAS  Google Scholar 

  19. Sowa, Y. and Sakai, T. (2000) Butyrate as a model for “gene-regulating chemoprevention and chemotherapy.” Biofactors 12, 283–287.

    Article  CAS  Google Scholar 

  20. Lamotte, D., Eon-Duval, A., Acerbis, G., Distefano, G., Monaco, L., Soria, M., et al. (1997) Controlling the glycosylation of recombinant proteins expressed in animal cells by genetic and physiological engineering, in Animal Cell Technology (Carrondo, M. J. T., ed.) Kluwer Academic Publishers, The Netherlands, pp. 761–765.

    Chapter  Google Scholar 

  21. Murray, K. and Dickson, A. J. (1997) Dichloroacetate inhibits glutamine oxidation by decreasing pyruvate availability for transamination. Metabolism 46, 268–472.

    Article  CAS  Google Scholar 

  22. Gu, X. and Wang, D. I. (1998) Improvement of interferon-gamma sialylation in Chinese hamster ovary cell culture by feeding of N-acetylmannosamine. Biotechnol. Bioeng. 58, 642–648.

    Article  CAS  Google Scholar 

  23. Green, N. H., Hooker, A. D., James, D. C., Baines, A. J., Strange, P. G., Jenkins, N., et al. (1997) Control of interferon-gamma glycosylation by the addition of defined lipid supplements to batch cultures of recombinant chinese hamster ovary cells, in Animal Cell Technology, Basic & Applied Aspects, Vol 8. (Funatsu, A., ed.) Kluwer Academic Publishers, The Netherlands, pp. 339–345.

    Chapter  Google Scholar 

  24. Monaco, L., Marc, A., Eon-Duval, A., Acerbis, G., Distefano, G., Lamotte, D., et al. (1996) Genetic engineering of α2,6-sialyltransferase in recombinant CHO cells and its effects on the sialylation of recombinant interferon-y. Cytotechnology 22, 197–203.

    Article  CAS  Google Scholar 

  25. Lund, J., Tanaka, T., Takahashi, N., Sarmay, G., Arata, Y., and Jefferis, R. (1990) A protein structural change in aglycosylated IgG3 correlates with loss of huFcyRI and huFcγRII binding and/or activation. Mol. Immunol. 27, 1145–1153.

    Article  CAS  Google Scholar 

  26. Sarmay, G., Lund, J., Rozsnyay, Z., Gergely, J., and Jefferis, R. (1992) Mapping and comparison of the interaction sites on the Fc region of IgG responsible for triggering antibody dependent cellular cytotoxicity (ADCC) through different types of human Fc receptor. Mol. Immunol. 29, 633–639.

    Article  CAS  Google Scholar 

  27. Morgan, A., Jones, N. D., Nesbitt, A. M., Chaplin, L., Bodmer, M. W., and Emtage, J. S. (1995) The N-terminal end of the CH2 domain of chimeric human IgG1 anti-HLA-DR is necessary for Clq, FcγRI and FcγRII binding. Immunology 86, 319–324.

    CAS  Google Scholar 

  28. Crocker, P. R., Hartnell, A., Munday, J., and Nath, D. (1997) The potential role of sialoadhesin as a macrophage recognition molecule in health and disease. Glycoconj. J. 14, 601–609.

    Article  CAS  Google Scholar 

  29. Bragonzi, A., Distefano, G., Buckberry, L. D., Acerbis, G., Foglieni, C., Lamotte, D., et al. (2000) A new Chinese hamster ovary cell line expressing alpha-2,6-sialyltransferase used as universal host for the production of human-like sialylated recombinant glycoproteins. Biochim. Biophys. Acta 1474, 273–282.

    Article  CAS  Google Scholar 

  30. Lee, E. U., Roth, J., and Paulsen, J. C. (1989) Alteration of terminal glycosylation sequences on N-linked oligosaccharides of chinese hamster ovary cells by expression of β-galactoside y2,6-sialyltransferase. J. Biol. Chem. 264, 13,848–13,855.

    Google Scholar 

  31. Monaco, L., Tagliabue, R., Soria, M. R., and Uhlen, M. (1994) An in vitro amplification approach for the expression of recombinant proteins in mammalian cells. Biotechnol. Appl. Biochem. 20, 157–171.

    CAS  Google Scholar 

  32. Mattox, S., Walrath, K., Ceiler, D., Smith, D. F., and Cummings, R. D. (1992) A solid-phase assay for the activity of CMPNeuAc-Gal β1,4G1cNAc-R α2,6-sialyltransferase. Anal. Biochem. 206, 430–436.

    Article  CAS  Google Scholar 

  33. Lipsky, N. G. and Pagano, R. E. (1985). A vital stain for the Golgi apparatus. Science 228, 745–747.

    Article  CAS  Google Scholar 

  34. Marks, D. L., Wu, K., Paul, P., Kamisaka, Y., Watanabe, R., and Pagano, R. E. (1999) Oligomerization and topology of the Golgi membrane protein glucosylceramide synthase. J. Biol. Chem. 274, 451–456.

    Article  CAS  Google Scholar 

  35. Hayter, P. M., Curling, E. M., Gould, M. L., Baines, A. J., Jenkins, N., Salmon, I., et al. (1993) The effect of dilution rate on CHO cell physiology and recombinant interferon-g production in glucose-limited chemostat cultures. Biotechnol. Bioeng. 42, 1077–1085.

    Article  CAS  Google Scholar 

  36. Fukuda, M. and Kobata, A. (1994) Glycobiology, A Practical Approach, IRL Press, Oxford, UK.

    Google Scholar 

  37. Waldburger, J. M., Masternak, K., Muhlethaler-Mottet, A., Villard, J., Peretti, M, Landmann, S., et al. (2000) Lessons from the bare lymphocyte syndrome: molecular mechanisms regulating MHC class II expression. Immunol. Rev. 178, 148–165.

    Article  CAS  Google Scholar 

  38. Kaufman, R. J., Wasley, L. C., Spiliotes, A. J., Gossels, S. D., Latt, S. A., Larsen, G. R., et al. (1985) Coamplification and coexpression of human tissue-type plasminogen activator and murine dihydrofolate reductase sequences in Chinese hamster ovary cells. Mol. Cell Biol. 5, 1750–1759.

    CAS  Google Scholar 

  39. Weikert, S., Papac, D., Briggs, J., Cowfer, D., Tom, S., Gawlitzek, M., et al. (1999) Engineering chinese hamster ovary cells to maximize sialic acid content of recombinant glycoproteins. Nat. Biotechnol. 17, 1116–1121.

    Article  CAS  Google Scholar 

  40. Shields, R. L., Namenuk, A. K., Hong, K., Meng, Y. G., Rae, J., Briggs, J., et al. (2000) High resolution mapping of the binding site on human IgG1 for FcγRI, FcγRII, FcyRIII and FcRn and design of IgG1 variants with improved binding to FcyR. J. Biol. Chem. 276, 6591–6604.

    Article  Google Scholar 

  41. Sondermann, P., Huber, R., Oosthuizen, V., and Jacob, U. (2000) A structural basis for immune complex recognition: the 3.2Å crystal structure of the human IgG1 Fc-fragment-FcγRIII complex. Nature 406, 267–273.

    Article  CAS  Google Scholar 

  42. Lund, J., Takahashi, N., Popplewell, A., Goodall, M., Pound, J. D., Tyler, R., King D. J., and Jefferis, R. (2000) Expression and characterization of truncated forms of humanized L243 IgG1 : architectural features can influence synthesis of its oligosaccharide chains and affect superoxide production triggered through human Fcy receptor I. Eur. J. Biochem. 267, 7246–7256.

    Article  CAS  Google Scholar 

  43. Jassal, R., Jenkins, N., Charlwood, J., Camilleri, P., Jefferis, R., and Lund, J. (2001) Sialylation of human IgG-Fc carbohydrate by transfected rat alpha2,6-sialyltransferase. Biochem. Biophys. Res. Commun. 286, 243–249.

    Article  CAS  Google Scholar 

  44. Deisenhofer, J. (1981) Crystallographic refinement and atomic models of a human Fc fragment and its complex with fragment B of protein A from Staphylococcus aureus at 2.9- and 2.8Å resolution. Biochemistry 20, 2361–2370.

    Article  CAS  Google Scholar 

  45. Hoffmann, A., Nimtz, M., Wurster, U., and Conradt, H. S. (1994) Carbohydrate structures of β-trace protein from human cerebrospinal fluid- evidence for brain-type N-glycosylation. J. Neurochem. 63, 2185–2196.

    Article  CAS  Google Scholar 

  46. Tsuchiya, N., Endo, T., Matsuta, K., Yoshinoya, S., Aikawa, T., Kosuge, E., et al. (1989) Effects of galactose depletion from oligosaccharide chains on immunological activities of human IgG. J. Rheumatol. 16, 285–290.

    CAS  Google Scholar 

  47. Lund, J., Takahashi, N., Pound, J. D., Nakagawa, H., Goodall, M., and Jefferis, R. (1995). Oligosaccharide-protein interactions in IgG can modulate recognition by Fcy receptors. FASEB J. 9, 115–119.

    CAS  Google Scholar 

  48. Mimura, Y., Church, S., Ghirlando, R., Ashton, P. R., Dong, S., Goodall, M., et al. (2000). The influence of glycosylation on the thermal stability and effector function expression of human IgG1-Fc: properties of a series of human glycoforms. Molec. Immunol. 37, 697–706.

    Article  CAS  Google Scholar 

  49. Umana, P., Jean-Mairet, J., Moudry, R., Amstutz, H., and Bailey, J. E. (1999). Engineered glycoforms of an antineuroblastoma IgG1 with optimized antibody-dependent cellular cytotoxic activity. Nat. Biotechnol. 17, 176–180.

    Article  CAS  Google Scholar 

Download references

Authors
  1. Nigel Jenkins
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Eli Lilly and Company, Indianapolis, IN, USA

    Victor A. Vinci PhD

  2. Dow AgroSciences, Indianapolis, IN, USA

    Sarad R. Parekh PhD

Rights and permissions

Reprints and permissions

Copyright information

© 2003 Springer Science+Business Media New York

About this chapter

Cite this chapter

Jenkins, N. (2003). Analysis and Manipulation of Recombinant Glycoproteins Manufactured in Mammalian Cell Culture. In: Vinci, V.A., Parekh, S.R. (eds) Handbook of Industrial Cell Culture. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59259-346-0_1

Download citation

  • DOI: https://doi.org/10.1007/978-1-59259-346-0_1

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61737-315-2

  • Online ISBN: 978-1-59259-346-0

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation