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Cited by 127Monoclonal and Polyclonal Antibodies ResearchGlycosylation and Glycoproteins ResearchProtein purification and stability

Quantitative evaluation of fucose reducing effects in a humanized antibody on Fcγ receptor binding and antibody-dependent cell-mediated cytotoxicity activities

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Chung, Shan, Quarmby, Valerie, Gao, Xiaoying, Ying, Yong, Lin, Linda, Reed, Chae, Fong, Chris, Lau, Wendy +4 more·2012·mAbs
DOI 10.4161/mabs.19941PMC PMC3355491
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Abstract

The presence or absence of core fucose in the Fc region N-linked glycans of antibodies affects their binding affinity toward FcγRIIIa as well as their antibody-dependent cell-mediated cytotoxicity (ADCC) activity. However, the quantitative nature of this structure-function relationship remains unclear. In this study, the in vitro biological activity of an afucosylated anti-CD20 antibody was fully characterized. Further, the effect of fucose reduction on Fc effector functions was quantitatively evaluated using the afucosylated antibody, its "regular" fucosylated counterpart and a series of mixtures containing varying proportions of "regular" and afucosylated materials. Compared with the "regular" fucosylated antibody, the afucosylated antibody demonstrated similar binding interactions with the target antigen (CD20), C1q and FcγRIa, moderate increases in binding to FcγRIIa and IIb, and substantially increased binding to FcγRIIIa. The afucosylated antibodies also showed comparable complement-dependent cytotoxicity activity but markedly increased ADCC activity. Based on EC 50 values derived from dose-response curves, our results indicate that the amount of afucosylated glycan in antibody samples correlate with both FcγRIIIa binding activity and ADCC activity in a linear fashion. Furthermore, the extent of ADCC enhancement due to fucose depletion was not affected by the FcγRIIIa genotype of the effector cells.

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  • This increased affinity allows better therapeutic activity of Mabs used in cancer therapy, by triggering the antibody-dependent-cell-mediated cytotoxicity (ADCC) pathway.

    “This increased affinity allows better therapeutic activity of Mabs used in cancer therapy, by triggering the antibody-dependent-cell-mediated cytotoxicity (ADCC) pathway (Niwa et al., 2005;Iida et al., 2006;Chung et al., 2012).”

    From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Chung et al., 2012

    The ADCC assay section reports that increased FcγRIIIa binding appeared to produce a proportional increase in ADCC activity. The Discussion also reports better responses to cancer antibodies in patients with a higher-affinity receptor variant. But the paper does not establish that increasing an antibody’s affinity itself improves clinical therapeutic outcomes; its direct antibody findings concern binding and ADCC assays. (LLM, conf=0.84)

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Cited by

Sorted by citations

Authors
Chung, S., Quarmby, V., Gao, X., Ying, Y., Lin, L., Reed, C., Fong, C., Lau, W., Qiu, Z. J., Shen, A., Vanderlaan, M., & Song, A.
Affiliations
Genentech (US)
Corresponding author
Chung, Shan
ORCID
Venue
mAbs
Publisher
Informa UK Limited
Published
2012-05
Volume
4
Issue
3
Pages
326-340
ISSN
  • 1942-0862 (ISSN-L)
  • 1942-0870
DOI
10.4161/mabs.19941
PMC
PMC3355491
OpenAlex
W2078353854
Cited by
127
References
2
Citations by year
2026: 6 · 2025: 2 · 2024: 11 · 2023: 5 · 2022: 14 · 2021: 12 · 2020: 13 · 2019: 11 · 2018: 12 · 2017: 9 · 2016: 9 · 2015: 11 · 2014: 5 · 2013: 7
FWCI
5.55
Type
journal article
Language
en
Open access
Locations
  • mAbs (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
  • 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
afucosylated antibody, core fucose, FcγRIIIa, Fcγ receptor binding, antibody-dependent cellular cytotoxicity, anti-CD20 monoclonal antibody, structure-function relationship, complement-dependent cytotoxicity, FcγRIIa, FcγRIIB, Fc effector functions
MeSH terms
Animals, Antibody Affinity, Antibody-Dependent Cell Cytotoxicity, Fucose, Humans, Oxidation-Reduction, Protein Binding, Protein Processing, Post-Translational, Receptors, IgG, Antigens, CD20, Quantitative Structure-Activity Relationship, Antibodies, Monoclonal, Humanized

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