Cited by 207Glycosylation and Glycoproteins ResearchViral Infectious Diseases and Gene Expression in InsectsCarbohydrate Chemistry and Synthesis

The DOI matches its Crossref record; 44 of the 44 references it cites were found in a registry or an index.

Glycomics Profiling of Chinese Hamster Ovary Cell Glycosylation Mutants Reveals N-Glycans of a Novel Size and Complexity

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Abstract

Identifying biological roles for mammalian glycans and the pathways by which they are synthesized has been greatly facilitated by investigations of glycosylation mutants of cultured cell lines and model organisms. Chinese hamster ovary (CHO) glycosylation mutants isolated on the basis of their lectin resistance have been particularly useful for glycosylation engineering of recombinant glycoproteins. To further enhance the application of these mutants, and to obtain insights into the effects of altering one specific glycosyltransferase or glycosylation activity on the overall expression of cellular glycans, an analysis of the N-glycans and major O-glycans of a panel of CHO mutants was performed using glycomic analyses anchored by matrix-assisted laser desorption ionization-time of flight/time of flight mass spectrometry. We report here the complement of the major N-glycans and O-glycans present in nine distinct CHO glycosylation mutants. Parent CHO cells grown in monolayer versus suspension culture had similar profiles of N- and O-GalNAc glycans, although the profiles of glycosylation mutants Lec1, Lec2, Lec3.2.8.1, Lec4, LEC10, LEC11, LEC12, Lec13, and LEC30 were consistent with available genetic and biochemical data. However, the complexity of the range of N-glycans observed was unexpected. Several of the complex N-glycan profiles contained structures of m/z approximately 13,000 representing complex N-glycans with a total of 26 N-acetyllactosamine (Gal beta1-4GlcNAc)(n) units. Importantly, the LEC11, LEC12, and LEC30 CHO mutants exhibited unique complements of fucosylated complex N-glycans terminating in Lewis(x) and sialyl-Lewis(x) determinants. This analysis reveals the larger-than-expected complexity of N-glycans in CHO cell mutants that may be used in a broad variety of functional glycomics studies and for making recombinant glycoproteins.

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  1. Stanley, 1989, Mol. Cell. Biol., 9, 377

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Metadata

Authors
North, S. J., Huang, H.-H., Sundaram, S., Jang-Lee, J., Etienne, A. T., Trollope, A., Chalabi, S., Dell, A., Stanley, P., & Haslam, S. M.
Affiliations
  • Imperial College London (GB)
  • Albert Einstein College of Medicine (US)
Corresponding author
  • Stanley, Pamela
  • Haslam, Stuart M.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2010-02
Volume
285
Issue
8
Pages
5759-5775
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2033604747
Cited by
207
References
44
Citations by year
2026: 5 · 2025: 7 · 2024: 8 · 2023: 6 · 2022: 10 · 2021: 12 · 2020: 11 · 2019: 15 · 2018: 17 · 2017: 16 · 2016: 8 · 2015: 11 · 2014: 21 · 2013: 17 · 2012: 13
FWCI
5.78
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by-nc)
  • PubMed (publishedVersion)
Funders
  • Wellcome Trust
  • National Institutes of Health
  • Directorate for Biological Sciences
  • U.S. Public Health Service (NCI RO1-36434)
  • Biotechnology and Biological Sciences Research Council (BB/F008309/1, B19088)
  • National Cancer Institute
  • National Institute of General Medical Sciences (GM 62116, U54GM062116)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
  • Physical Sciences › Chemistry › Organic Chemistry › Carbohydrate Chemistry and Synthesis
Keywords
glycosylation mutants, CHO cells, N-glycans, O-glycans, glycomics, lectin resistance, glycosyltransferases, N-acetyllactosamine, fucosylated N-glycans, CD15, sialyl Lewis X
MeSH terms
Animals, Carbohydrate Sequence, Cricetulus, Glycosylation, Cricetinae, Mutation, Polysaccharides, CHO Cells

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