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Cited by 41Viral Infectious Diseases and Gene Expression in InsectsGlycosylation and Glycoproteins ResearchProtein purification and stability

A theoretical estimate for nucleotide sugar demand towards Chinese Hamster Ovary cellular glycosylation

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del Val, Ioscani Jimenez, Polizzi, Karen M., Kontoravdi, Cleo·2016·Scientific Reports
DOI 10.1038/srep28547PMC PMC4921913
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Abstract

Glycosylation greatly influences the safety and efficacy of many of the highest-selling recombinant therapeutic proteins (rTPs). In order to define optimal cell culture feeding strategies that control rTP glycosylation, it is necessary to know how nucleotide sugars (NSs) are consumed towards host cell and rTP glycosylation. Here, we present a theoretical framework that integrates the reported glycoproteome of CHO cells, the number of N-linked and O-GalNAc glycosylation sites on individual host cell proteins (HCPs), and the carbohydrate content of CHO glycosphingolipids to estimate the demand of NSs towards CHO cell glycosylation. We have identified the most abundant N-linked and O-GalNAc CHO glycoproteins, obtained the weighted frequency of N-linked and O-GalNAc glycosites across the CHO cell proteome, and have derived stoichiometric coefficients for NS consumption towards CHO cell glycosylation. By combining the obtained stoichiometric coefficients with previously reported data for specific growth and productivity of CHO cells, we observe that the demand of NSs towards glycosylation is significant and, thus, is required to better understand the burden of glycosylation on cellular metabolism. The estimated demand of NSs towards CHO cell glycosylation can be used to rationally design feeding strategies that ensure optimal and consistent rTP glycosylation.

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  • Nucleotide sugar donors (NSDs) serve as essential substrates for N-linked glycosylation, a critical quality attribute that strongly influences the safety, efficacy, and pharmacokinetics of recombinant protein products.

    “Specifically, nucleotide sugar donors (NSDs) serve as essential substrates for N -linked glycosylation, a critical quality attribute that strongly influences the safety, efficacy, and pharmacokinetics of recombinant protein products [11,12].”

    From Soft sensing of intracellular states for CHO cell bioprocessing with Ensemble Kalman Filters (2026) · cites this paper as [11]

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Cited by

Sorted by citations

Authors
del Val, I. J., Polizzi, K. M., & Kontoravdi, C.
Affiliations
Imperial College London (GB)
ORCID
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2016-06-27
Volume
6
Issue
1
Pages
28547-28547
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/srep28547
PMC
PMC4921913
OpenAlex
W2469312676
Cited by
41
References
4
Citations by year
2026: 4 · 2025: 3 · 2024: 4 · 2023: 1 · 2022: 5 · 2021: 5 · 2020: 6 · 2019: 2 · 2018: 5 · 2017: 4 · 2016: 1
FWCI
1.71
Type
journal article
Language
en
Open access
Licence
Locations
  • Scientific Reports (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Spiral (Imperial College London) (submittedVersion, cc-by)
Funders
  • Research Councils UK
  • Engineering and Physical Sciences Research Council (BB/I017011/1)
  • Biotechnology and Biological Sciences Research Council (BB/I017011/1)
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
sugar nucleotides, recombinant therapeutic proteins, N-glycosylation, O-GalNAc glycosylation, glycoproteome, glycosphingolipids, stoichiometric coefficients, host cell proteins, CHO cells
MeSH terms
Sugars, Animals, Carbohydrates, Cricetulus, Glycoproteins, Glycosylation, Cricetinae, Nucleotides, Recombinant Proteins, CHO Cells, Cell Culture Techniques, Proteome

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