Glycosylation analysis of an aggregated antibody produced by Chinese hamster ovary cells in bioreactor culture

Masayoshi Onitsuka, Akira Kawaguchi, Ryutaro Asano, Izumi Kumagai, Kohsuke Honda, Hisao Ohtake, Takeshi Omasa

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

N-Glycosylation of therapeutic antibodies contributes not only to their biological function, but also to their stability and tendency to aggregate. Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates.

Original languageEnglish
Pages (from-to)639-644
Number of pages6
JournalJournal of Bioscience and Bioengineering
Volume117
Issue number5
DOIs
Publication statusPublished - 2014 May

Keywords

  • Antibody aggregation
  • Antibody production
  • Bispecific diabody
  • Cell culture
  • Chinese hamster ovary cell
  • Glycosylation heterogeneity

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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