Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity

Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. The resulting antibody demonstrated enhanced antibody-dependent cell-mediated...

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Bibliographic Details
Published inmAbs Vol. 8; no. 8; pp. 1498 - 1511
Main Authors Calow, Jenny, Behrens, Anna-Janina, Mader, Sonja, Bockau, Ulrike, Struwe, Weston B, Harvey, David J, Cormann, Kai U, Nowaczyk, Marc M, Loser, Karin, Schinor, Daniel, Hartmann, Marcus W W, Crispin, Max
Format Journal Article
LanguageEnglish
Published United States Taylor & Francis 16.11.2016
Subjects
Animals
Antibody-Dependent Cell Cytotoxicity - immunology
Bioreactors - microbiology
Glycosylation
Humans
Mice
Rituximab - biosynthesis
Rituximab - chemistry
Rituximab - immunology
Tetrahymena thermophila
recombinant expression
monoclonal antibody
ADCC
N-glycosylation pattern
rituximab
ciliate
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Summary:Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. The resulting antibody demonstrated enhanced antibody-dependent cell-mediated cytotoxicity, which we attribute to unusual N-linked glycosylation. Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. In addition, terminal glucose structures were identified that match biosynthetic precursors of human glycosylation. Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity.
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Supplemental data for this article can be accessed on the publisher's website.
These authors share first co-authorship.
ISSN:1942-0862
1942-0870
DOI:10.1080/19420862.2016.1228504