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Structural and Functional Differences Between Glycosylated and Non-glycosylated Forms of Human Interferon-β (IFN-β)
Abstract
Purpose Two recombinant IFN-beta products have been approved for the treatment of multiple sclerosis, a glycosylated form with the predicted natural amino acid sequence (IFN-beta-1a) and a non-glycosylated form that has a Met-1 deletion and a Cys-17 to Ser mutation (IFN-beta-1b). The structural basis for activity differences between IFN-beta-1a and IFN-beta-1b, is determined. Methods In vitro antiviral, antiproliferative and immunomodulatory assays were used to directly compare the two IFN-beta products. Size exclusion chromatography (SEC), SDS-PAGE, thermal denaturation, and X-ray crystallography were used to examine structural differences. Results IFN-beta-1a was 10 times more active than IFN-beta-1b with specific activities in a standard antiviral assay of 20 x 10(7) IU/mg for IFN-beta-1a and 2 x 10(7) IU/mg for IFN-beta-1b. Of the known structural differences between IFN-beta-1a and IFN-beta-1b, only glycosylation affected in vitro activity. Deglycosylation of IFN-beta-1a produced a decrease in total activity that was primarily caused by the formation of an insoluble disulfide-linked IFN precipitate. Deglycosylation also resulted in an increased sensitivity to thermal denaturation. SEC data for IFN-beta-1b revealed large, soluble aggregates that had reduced antiviral activity (approximated at 0.7 x 10(7) IU/mg). Crystallographic data for IFN-beta-1a revealed that the glycan formed H-bonds with the peptide backbone and shielded an uncharged surface from solvent exposure. C…
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“Studies have demonstrated that proper glycosylation profiles can improve recombinant protein properties such as increase their stability and half-life in blood circulation and decrease their immunogenicity (Ashwell and Harford, 1982;Runkel et al., 1998;Ghaderi et al., 2010;Ghaderi et al., 2012;Jefferis, 2016a;Jefferis, 2016b;Kuriakose et al., 2016).”
From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Runkel et al., 1998
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Metadata
- Authors
- Runkel, L., Meier, W., Pepinsky, R. B., Karpusas, M., Whitty, A., Kimball, K., Brickelmaier, M., Muldowney, C., Jones, W., & Goelz, S. E.
- Affiliations
- Biogen (United States) (US)
- Corresponding author
- Runkel, Laura
- ORCID
- Whitty, Adrian
- Venue
- Pharmaceutical Research
- Published
- 1998-04
- Volume
- 15
- Issue
- 4
- Pages
- 641-649
- ISSN
- 0724-8741 (ISSN-L)
- 1573-904X
- OpenAlex
- W329292291
- Cited by
- 304
- References
- 34
- Citations by year
- 2026: 4 · 2025: 4 · 2024: 4 · 2023: 5 · 2022: 9 · 2021: 10 · 2020: 11 · 2019: 8 · 2018: 10 · 2017: 8 · 2016: 16 · 2015: 10 · 2014: 16 · 2013: 16 · 2012: 13
- FWCI
- 3.46
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- Locations
- Pharmaceutical Research (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › interferon and immune responses
- Health Sciences › Medicine › Clinical Neurology › Multiple Sclerosis Research Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
- Keywords
- interferon-beta, glycosylation, recombinant interferon, multiple sclerosis, antiviral activity, protein stability, thermal denaturation, protein aggregation, disulfide bonds, protein structure
- MeSH terms
- Interferon beta-1a, Interferon beta-1b, Adjuvants, Immunologic, Antiviral Agents, Base Sequence, Cell Division, Cell Line, Cysteine, Encephalomyocarditis virus, Glycosylation, Humans, Major Histocompatibility Complex, Molecular Sequence Data, Mutation, Recombinant Proteins, Serine, Interferon-beta, Cardiovirus Infections
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