Supplementing glycosylation: A review of applying nucleotide-sugar precursors to growth medium to affect therapeutic recombinant protein glycoform distributions
Open accessAbstract
Glycosylation is a critical quality attribute (CQA) of many therapeutic proteins, particularly monoclonal antibodies (mAbs), and is a major consideration in the approval of biosimilar biologics due to its effects to therapeutic efficacy. Glycosylation generates a distribution of glycoforms, resulting in glycoproteins with inherent molecule-to-molecule heterogeneity, capable of activating (or failing to activate) different effector functions of the immune system. Glycoforms can be affected by the supplementation of nucleotide-sugar precursors, and related components, to culture growth medium, affecting the metabolism of glycosylation. These supplementations has been demonstrated to increase nucleotide-sugar intracellular pools, and impact glycoform distributions, but with varied results. These variations can be attributed to five key factors: Differences between cell platforms (enzyme/transporter expression levels); differences between recombinant proteins produced (glycan-site accessibility); the fermentation and sampling timeline (glucose availability and exoglycosidase accumulation); glutamine levels (affecting ammonia levels, which impact Golgi pH, as well as UDP-GlcNAc pools); and finally, a lack of standardized metrics for observing shifts in glycoform distributions (glycosylation indices) across different experiments. The purpose of this review is to provide detail and clarity on the state of the art of supplementation strategies for nucleotide-sugar precursors for affecting glycosylation in cell culture processes, and to apply glycosylation indices for standardized comparisons across the field.
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UDP-Gal is the immediate intracellular precursor for terminal galactosylation of N-linked glycans.
“UDP-Gal is the immediate intracellular precursor for terminal galactosylation of N -linked glycans [49].”
From Soft sensing of intracellular states for CHO cell bioprocessing with Ensemble Kalman Filters (2026) · cites this paper as [49]
Section 2.1 (p. 6) describes nucleotide-sugars as glycosylation substrates, and Section 3.3.2 (p. 19) reports that galactose precursor feeding increases galactosylation. Figure 1 (p. 48) labels a UDP-Gal transporter, but the available text does not explicitly identify UDP-Gal as the immediate intracellular precursor for terminal galactosylation of N-linked glycans. (LLM, conf=0.80)
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InfluentialSoft sensing of intracellular states for CHO cell bioprocessing with Ensemble Kalman Filters
Yu, L., del Rio Chanona, A., & Kontoravdi, C. (2026) · Journal of Process Control · doi:10.1016/j.jprocont.2026.103875
- Authors
- Blondeel, E. J. M., & Aucoin, M. G.
- Affiliations
- University of Waterloo (CA)
- Corresponding author
- Aucoin, Marc G.
- ORCID
- Aucoin, Marc G.
- Venue
- Biotechnology Advances
- Publisher
- Elsevier BV
- Published
- 2018-09
- Volume
- 36
- Issue
- 5
- Pages
- 1505-1523
- ISSN
- 0734-9750 (ISSN-L)
- OpenAlex
- W2809077322
- Cited by
- 46
- References
- 10
- Citations by year
- 2026: 8 · 2025: 4 · 2024: 4 · 2023: 5 · 2022: 4 · 2021: 7 · 2020: 8 · 2019: 4 · 2018: 1
- FWCI
- 0.50
- Type
- journal article
- Language
- en
- Open access
- green
- hdl.handle.net
- Licence
- cc-by-nc-nd
- www.elsevier.com/tdm/userlicense/1.0/ (tdm)
- Locations
- Biotechnology Advances (publishedVersion)
- PubMed (publishedVersion)
- UWSpace (University of Waterloo) (submittedVersion, cc-by-nc-nd)
- Funders
- NSERC Strategic Network Grant (MabNet - NETGP 380070-08)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Keywords
- glycosylation, monoclonal antibodies, culture medium supplementation, biosimilars, UDP-GlcNAc, exoglycosidases, glutamine
- MeSH terms
- Animals, Antibodies, Monoclonal, Cricetulus, Culture Media, Glycosylation, Cricetinae, Nucleotides, Recombinant Proteins, CHO Cells, Bioreactors
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