期刊
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
卷 94, 期 1, 页码 69-80出版社
SPRINGER
DOI: 10.1007/s00253-011-3814-1
关键词
Chinese hamster ovary (CHO) cell; Antibody production; Glycosylation control; Sialylation; Beta-galactosyl alpha-2,6 sialyltransferase (ST6Gal I); Bispecific diabody
资金
- New Energy and Industrial Technology Development Organization (NEDO) of Japan
- National Institute of Biomedical Innovation (NIBIO)
- Japan Society for the Promotion of Science (JSPS)
- Grants-in-Aid for Scientific Research [23686118, 23360371] Funding Source: KAKEN
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.
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