Modulation of therapeutic antibody effector functions by glycosylation engineering: Influence of Golgi enzyme localization domain and co‐expression of heterologous β1, 4‐N‐acetylglucosaminyltransferase III and Golgi α‐mannosidase II
Abstract
The effector functions elicited by IgG antibodies strongly depend on the carbohydrate moiety linked to the Fc region of the protein. Therefore several approaches have been developed to rationally manipulate these glycans and improve the biological functions of the antibody. Overexpression of recombinant β1,4‐N‐acetylglucosaminyltransferase III (GnT‐III) in production cell lines leads to antibodies enriched in bisected oligosaccharides. Moreover, GnT‐III overexpression leads to increases in non‐fucosylated and hybrid oligosaccharides. Such antibody glycovariants have increased antibody‐dependent cellular cytotoxicity (ADCC). To explore a further variable besides overexpression of GnT‐III, we exchanged the localization domain of GnT‐III with that of other Golgi‐resident enzymes. Our results indicate that chimeric GnT‐III can compete even more efficiently against the endogenous core α1,6‐fucosyltransferase (α1,6‐FucT) and Golgi α‐mannosidase II (ManII) leading to higher proportions of bisected non‐fucosylated hybrid glycans (“Glyco‐1” antibody). The co‐expression of GnT‐III and ManII led to a similar degree of non‐fucosylation as that obtained for Glyco‐1, but the majority of the oligosaccharides linked to this antibody (“Glyco‐2”) are of the complex type. These glycovariants feature strongly increased ADCC activity compared to the unmodified antibody, while Glyco‐1 (hybrid‐rich) features reduced complement‐dependent cytotoxicity (CDC) compared to Glyco‐2 or unmodified antibody. We show that apart from GnT‐III overexpression, engineering of GnT‐III localization is a versatile tool to modulate the biological activities of antibodies relevant for their therapeutic application. © 2006 Wiley Periodicals, Inc.
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Since expression of GnTIII blocks fucosylation, CHO cells were engineered with both GnTIII and Golgi mannosidase II (ManII) for IgG production.
“Since expression of the β1,4-mannosyl-glycoprotein 4-β-N-acetylglucosaminyltransferase III (GnTIII) blocks fucosylation, CHO cells were engineered with both GnTIII and Golgi mannosidase II (ManII) for IgG production (Davies et al., 2001;Ferrara et al., 2006).”
From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Ferrara et al., 2006
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- Authors
- Ferrara, C., Brünker, P., Suter, T., Moser, S., Püntener, U., & Umaña, P.
- Affiliations
- ETH Zurich (CH)
- Corresponding author
- Umaña, Pablo
- Venue
- Biotechnology and Bioengineering
- Publisher
- Wiley
- Published
- 2006-01-24
- Volume
- 93
- Issue
- 5
- Pages
- 851-861
- ISSN
- 0006-3592 (ISSN-L)
- 1097-0290
- OpenAlex
- W2152155460
- Cited by
- 305
- References
- 3
- Citations by year
- 2026: 5 · 2025: 7 · 2024: 10 · 2023: 5 · 2022: 14 · 2021: 15 · 2020: 11 · 2019: 18 · 2018: 18 · 2017: 17 · 2016: 24 · 2015: 15 · 2014: 21 · 2013: 16 · 2012: 19
- FWCI
- 7.60
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biotechnology and Bioengineering (publishedVersion)
- PubMed (publishedVersion)
- 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
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Keywords
- therapeutic antibodies, antibody glycosylation, Golgi alpha-mannosidase II, Fc glycosylation, antibody-dependent cellular cytotoxicity, complement-dependent cytotoxicity, antibody effector functions
- MeSH terms
- Animals, Antibody-Dependent Cell Cytotoxicity, B-Lymphocytes, Cell Line, Complement System Proteins, Cricetulus, Cytotoxicity, Immunologic, Genetic Vectors, Glycosylation, Golgi Apparatus, Cricetinae, Humans, Immunoglobulin G, Killer Cells, Natural, Oligosaccharides, Protein Processing, Post-Translational, Recombinant Fusion Proteins, Transfection, Protein Engineering, CHO Cells, N-Acetylgalactosaminyltransferases, Receptors, IgG, Antigens, CD20, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Protein Sorting Signals, alpha-Mannosidase, Cell Line, Tumor
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