Biotechnological Products and Process Engineering

Applied Microbiology and Biotechnology

, Volume 85, Issue 3, pp 535-542

First online:

Glycosylation pattern of humanized IgG-like bispecific antibody produced by recombinant CHO cells

  • Wook-Dong KimAffiliated withDepartment of Biotechnology, Graduate School of Engineering, Osaka University
  • , Miwako TokunagaAffiliated withDepartment of Biotechnology, Graduate School of Engineering, Osaka University
  • , Hiroyuki OzakiAffiliated withDepartment of Biotechnology, Graduate School of Engineering, Osaka University
  • , Takuya IshibashiAffiliated withToyobo Biologics Inc.
  • , Kohsuke HondaAffiliated withDepartment of Biotechnology, Graduate School of Engineering, Osaka University
  • , Hiroyuki KajiuraAffiliated withThe International Center for Biotechnology, Osaka University
  • , Kazuhito FujiyamaAffiliated withThe International Center for Biotechnology, Osaka University
  • , Ryutaro AsanoAffiliated withDepartment of Biomolecular Engineering, Graduate School of Engineering, Tohoku University
  • , Izumi KumagaiAffiliated withDepartment of Biomolecular Engineering, Graduate School of Engineering, Tohoku University
    • , Takeshi OmasaAffiliated withDepartment of Biotechnology, Graduate School of Engineering, Osaka University Email author 
    • , Hisao OhtakeAffiliated withDepartment of Biotechnology, Graduate School of Engineering, Osaka University

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Abstract

The glycosylation pattern of a humanized anti-EGFR×anti-CD3 bispecific single-chain diabody with an Fc portion (hEx3-scDb-Fc) produced by recombinant Chinese hamster ovary cells was evaluated and compared with those of a recombinant humanized anti-IL-8 antibody (IgG1) and human serum IgG. N-Linked oligosaccharide structures were estimated by two-dimensional high-performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. No sialylation was observed with purified hEx3-scDb-Fc and the anti-IL-8 antibody. From the analysis of neutral oligosaccharides, approximately more than 90% of the N-linked oligosaccharides of hEx3-scDb-Fc and the anti-IL-8 antibody were alpha-1,6-fucosylated. The galactosylated biantennary oligosaccharides comprise over 40% of the total N-linked oligosaccharides in both hEx3-scDb-Fc and the anti-IL-8 antibody. The fully galactosylated biantennary oligosaccharides from hEx3-scDb-Fc and the anti-IL-8 antibody accounted for only 10% of the N-linked; however, more than 20% of the N-linked oligosaccharides were fully galactosylated biantennary oligosaccharides in human serum IgG. The glycosylation pattern of hEx3-scDb-Fc was quite similar to that of the anti-IL-8 antibody.

Keywords

Bispecific diabody Bispecific IgG-like antibody Glycosylation Immunoglobulin G Chinese hamster ovary cells