Applied Microbiology and Biotechnology

, Volume 85, Issue 3, pp 535–542

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

  • Wook-Dong Kim
  • Miwako Tokunaga
  • Hiroyuki Ozaki
  • Takuya Ishibashi
  • Kohsuke Honda
  • Hiroyuki Kajiura
  • Kazuhito Fujiyama
  • Ryutaro Asano
  • Izumi Kumagai
  • Takeshi Omasa
  • Hisao Ohtake
Biotechnological Products and Process Engineering

DOI: 10.1007/s00253-009-2152-z

Cite this article as:
Kim, WD., Tokunaga, M., Ozaki, H. et al. Appl Microbiol Biotechnol (2010) 85: 535. doi:10.1007/s00253-009-2152-z

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 

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Wook-Dong Kim
    • 1
  • Miwako Tokunaga
    • 1
  • Hiroyuki Ozaki
    • 1
  • Takuya Ishibashi
    • 2
  • Kohsuke Honda
    • 1
  • Hiroyuki Kajiura
    • 3
  • Kazuhito Fujiyama
    • 3
  • Ryutaro Asano
    • 4
  • Izumi Kumagai
    • 4
  • Takeshi Omasa
    • 1
  • Hisao Ohtake
    • 1
  1. 1.Department of Biotechnology, Graduate School of EngineeringOsaka UniversitySuitaJapan
  2. 2.Toyobo Biologics Inc.OtsuJapan
  3. 3.The International Center for BiotechnologyOsaka UniversitySuitaJapan
  4. 4.Department of Biomolecular Engineering, Graduate School of EngineeringTohoku UniversitySendaiJapan

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