Journal Article

Production of non-fucosylated antibodies by co-expression of heterologous GDP-6-deoxy-<span class="smallCaps">d</span>-lyxo-4-hexulose reductase

Hans Henning von Horsten, Christiane Ogorek, Véronique Blanchard, Christian Demmler, Christoph Giese, Karsten Winkler, Matthias Kaup, Markus Berger, Ingo Jordan and Volker Sandig

in Glycobiology

Published on behalf of Society for Glycobiology

Volume 20, issue 12, pages 1607-1618
Published in print December 2010 | ISSN: 0959-6658
Published online July 2010 | e-ISSN: 1460-2423 | DOI: http://dx.doi.org/10.1093/glycob/cwq109
Production of non-fucosylated antibodies by co-expression of heterologous GDP-6-deoxy-d-lyxo-4-hexulose reductase

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All IgG-type antibodies are N-glycosylated in their Fc part at Asn-297. Typically, a fucose residue is attached to the first N-acetylglucosamine of these complex-type N-glycans. Antibodies lacking core fucosylation show a significantly enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) and an increased efficacy of anti-tumor activity. In cases where the clinical efficacy of an antibody is to some extent mediated by its ADCC effector function, afucosylated N-glycans could help to reduce dose requirement and save manufacturing costs. Using Chinese hamster ovary (CHO) cells as a model, we demonstrate here that heterologous expression of the prokaryotic enzyme GDP-6-deoxy-d-lyxo-4-hexulose reductase within the cytosol can efficiently deflect the fucose de novo pathway. Antibody-producing CHO cells that were modified in this way secrete antibodies lacking core fucose as demonstrated by MALDI-TOF mass spectrometry and HPAEC-PAD monosaccharide analysis. Engineering of the fucose de novo pathway has led to the construction of IgGs with a strongly enhanced ADCC effector function. The method described here should have broad practical applicability for the development of next-generation therapeutic antibodies.

Keywords: antibody; cell culture; core fucose; glycoengineering; sugar nucleotide metabolism

Journal Article.  7636 words.  Illustrated.

Subjects: Carbohydrates

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