Enhancement of sialylation on humanized IgG-like bispecific antibody by overexpression of α2,6-sialyltransferase derived from Chinese hamster ovary cells

Masayoshi Onitsuka, Wook Dong Kim, Hiroyuki Ozaki, Akira Kawaguchi, Kohsuke Honda, Hiroyuki Kajiura, Kazuhito Fujiyama, Ryutaro Asano, Izumi Kumagai, Hisao Ohtake, Takeshi Omasa

Research output: Contribution to journalArticlepeer-review

29 Citations (Scopus)

Abstract

Improvement of glycosylation is one of the most important topics in the industrial production of therapeutic antibodies. We have focused on terminal sialylation with alpha-2,6 linkage, which is crucial for anti-inflammatory activity. In the present study, we have successfully cloned cDNA of beta-galactosyl alpha-2,6 sialyltransferase (ST6Gal I) derived from Chinese hamster ovary (CHO) cells regardless of reports that stated this was not endogenously expressed in CHO cells. After expressing cloned ST6Gal I in Escherichia coli, the transferase activity was confirmed by HPLC and lectin binding assay. Then, we applied ST6Gal I to alpha-2,6 sialylation of the recombinant antibody; the ST6Gal I expression vector was transfected into the CHO cell line producing a bispecific antibody. The N-glycosylation pattern of the antibody was estimated by HPLC and sialidase digestion. About 70% of the total N-linked oligosaccharide was alpha-2,6 sialylated in the transfected cell line whereas no sialylation was observed in the non-transfected cell line. The improvement of sialylation would be of practical importance for the industrial production of therapeutic antibodies.

Original languageEnglish
Pages (from-to)69-80
Number of pages12
JournalApplied Microbiology and Biotechnology
Volume94
Issue number1
DOIs
Publication statusPublished - 2012 Apr

Keywords

  • Antibody production
  • Beta-galactosyl alpha-2,6 sialyltransferase (ST6Gal I)
  • Bispecific diabody
  • Chinese hamster ovary (CHO) cell
  • Glycosylation control
  • Sialylation

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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