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Mol Immunol. 2016 May;73:69-75. doi: 10.1016/j.molimm.2016.03.002. Epub 2016 Apr 6.

Fc-galactosylation modulates antibody-dependent cellular cytotoxicity of therapeutic antibodies.

Author information

1
Pharma Technical Development Analytics Biologics, Roche Diagnostics GmbH, Penzberg, Germany. Electronic address: marco.thomann@roche.com.
2
Pharma Biotech Manufacturing Penzberg, Roche Diagnostics GmbH, Penzberg, Germany.
3
Pharma Technical Development Analytics Biologics, Roche Diagnostics GmbH, Penzberg, Germany.
4
Analytical Development & Quality Control, Genentech Inc., South San Francisco, CA, USA.

Abstract

The therapeutic activity of monoclonal antibodies can involve immune cell mediated effector functions including antibody-dependent cellular cytotoxicity (ADCC), an activity that is modulated by the structure of Fc-glycans, and in particular the lack of core fucose. The heterogeneity of these glycostructures and the inherent variability of traditional PBMC-based in vitro ADCC assays, have made it challenging to quantitatively assess the impact of other glycostructures on ADCC activity. We applied a quantitative NK cell based assay to generate a database consisting of Fc-glycostructure and ADCC data from 54 manufacturing batches of a CHO-derived monoclonal antibody. Explorative analysis of the data indicated that, apart from afucosylation, galactosylation levels could influence ADCC activity. We confirmed this hypothesis by demonstrating enhanced ADCC upon enzymatic hypergalactosylation of four different monoclonal antibodies derived using standard CHO manufacturing processes. Furthermore we quantitatively compare the effects of galactosylation and afucosylation in the context of glycan heterogeneity and demonstrate that while galactose can influence ADCC activity, afucosylation remains the primary driver of this activity.

KEYWORDS:

ADCC; Afucosylation; Antibody; CDC; Effector function; Fc-glycan; Fucosylation; Galactosylation; IgG1; Natural killer cell

PMID:
27058641
DOI:
10.1016/j.molimm.2016.03.002
[Indexed for MEDLINE]
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