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N-Glycosylation of Recombinant Human Interferon-γ Produced in Different Animal Expression Systems

Bio/Technology volume 13, pages 592–596 (1995)Cite this article

Abstract

Recombinant human interferon-γ (IFN-γ) was expressed in Chinese hamster ovary cells, baculovirus-infected Sf9 insect cells and the mammary gland of transgenic mice. The N-linked carbohydrate populations associated with both Asn25 and Asn97 glycosylation sites were characterized by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) in combination with exoglycosidase array sequencing. A site-specific analysis of dual (2N) and single (IN) site-occupancy variants of IFN-γ derived from Chinese hamster ovary cells showed that N-glycans were predominantly of the complex bi- and triantennary type. Although Asn25-linked glycans were substituted with a core fucose residue, Asn97 in N-glycans were predominantly non-fucosylated, and truncated complex and high-mannose oligosaccha-ride chains were also evident. Transgenic mouse derived IFN-γ exhibited considerable site-specific variation in N-glycan structures. Asn97-linked carbohydrates were of the complex, core fucosylated type, Asn25-linked carbohydrates were mainly of the oligomannose type, with smaller proportions of hybrid and complex N-glycans. Carbohydrates associated with both glycosylation sites of IFN-γ from Sf9 insect cells were mainly tri-mannosyl core structures, with fucosylation confined to the ASn25 site. These data demonstrate the profound influence of host cell type and protein structure on the N-glycosylation of recombinant proteins.

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Author notes

  1. Nigel Jenkins

    Present address: Department of Biological Sciences, de Montfort University, The Gateway, Leicester, LE1 9BH, U.K.

Authors and Affiliations

  1. Research School of Biosciences, University of Kent, Canterbury, CT2 7NJ, U.K.

    David C. James, Robert B. Freedman, Olotu W. Ogonah, Barrie C. Rooney & Nigel Jenkins

  2. The Advanced Centre for Biochemical Engineering, Department of Chemical and Biochemical Engineering, University College London, Torrington Place, London, WC1E 7JE, U.K.

    David C. James & Michael Hoare

  3. Shemyakin Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya, 16/10, 117871 GSP, Moscow, V-437, Russia

    Oleg A. Larionov, Vasily N. Dobrovolsky & Oleg V. Lagutin

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Correspondence to David C. James.

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James, D., Freedman, R., Hoare, M. et al. N-Glycosylation of Recombinant Human Interferon-γ Produced in Different Animal Expression Systems. Nat Biotechnol 13, 592–596 (1995). https://doi.org/10.1038/nbt0695-592

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