Increased in vivo effector function of human IgG4 isotype antibodies through afucosylation
Open accessAbstract
For some antibodies intended for use as human therapeutics, reduced effector function is desired to avoid toxicities that might be associated with depletion of target cells. Since effector function(s), including antibody-dependent cell-mediated cytotoxicity (ADCC), require the Fc portion to be glycosylated, reduced ADCC activity antibodies can be obtained through aglycosylation of the human IgG1 isotype. An alternative is to switch to an IgG4 isotype in which the glycosylated antibody is known to have reduced effector function relative to glycosylated IgG1 antibody. ADCC activity of glycosylated IgG1 antibodies is sensitive to the fucosylation status of the Fc glycan, with both in vitro and in vivo ADCC activity increased upon fucose removal ("afucosylation"). The effect of afucosylation on activity of IgG4 antibodies is less well characterized, but it has been shown to increase the in vitro ADCC activity of an anti-CD20 antibody. Here, we show that both in vitro and in vivo activity of anti-CD20 IgG4 isotype antibodies is increased via afucosylation. Using blends of material made in Chinese hamster ovary (CHO) and Fut8KO-CHO cells, we show that ADCC activity of an IgG4 version of an anti-human CD20 antibody is directly proportional to the fucose content. In mice transgenic for human FcγRIIIa, afucosylation of an IgG4 anti-mouse CD20 antibody increases the B cell depletion activity to a level approaching that of the mIgG2a antibody.
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The same effect of afucosylated Fc domain glycans enhancing FcγRIIIa binding was observed for human IgG4.
“Recently, the same effect was observed for human IgG4 (Gong et al., 2016).”
From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Gong et al., 2016
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- Authors
- Gong, Q., Hazen, M., Marshall, B., Crowell, S. R., Ou, Q., Wong, A. W., Phung, W., Vernes, J.-M., Meng, Y. G., Tejada, M., Andersen, D., & Kelley, R. F.
- Affiliations
- Genentech (US)
- Corresponding author
- Kelley, Robert F.
- ORCID
- Venue
- mAbs
- Publisher
- Informa UK Limited
- Published
- 2016-05-24
- Volume
- 8
- Issue
- 6
- Pages
- 1098-1106
- ISSN
- 1942-0862 (ISSN-L)
- 1942-0870
- PMC
- PMC4968109
- OpenAlex
- W2403376704
- Cited by
- 26
- References
- 34
- Citations by year
- 2026: 2 · 2025: 2 · 2024: 4 · 2023: 4 · 2021: 4 · 2020: 3 · 2019: 2 · 2018: 3 · 2017: 2
- FWCI
- 2.03
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- 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 › Viral Infectious Diseases and Gene Expression in Insects
- Keywords
- IgG4 antibodies, afucosylation, Fc glycosylation, antibody-dependent cellular cytotoxicity, anti-CD20 monoclonal antibody, FcγRIIIa, B-cell depletion, therapeutic antibodies
- MeSH terms
- Animals, Antibodies, Monoclonal, Antibody-Dependent Cell Cytotoxicity, B-Lymphocytes, Blood, Cricetulus, Female, Fucose, Glycosylation, Cricetinae, Humans, Immunoglobulin G, Lymph Nodes, Lymphocyte Depletion, Mice, Inbred C57BL, Mice, Transgenic, Spleen, CHO Cells, Antigens, CD20, Mice
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