期刊
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
卷 477, 期 2, 页码 299-304出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.abb.2008.05.021
关键词
endo-beta-N-acetylglucosaminidase; endo-M; monoclonal antibody; N-linked complex-type sialo-oligosaccharide; transglycosylation; neoglycolipid
资金
- Ministry of Education, Science, Sports and Culture of Japan [10556079]
- Japan Society for the Promotion of Science
- Mizutani Foundation for Glycoscience
- Grants-in-Aid for Scientific Research [10556079] Funding Source: KAKEN
We attempted to obtain the monoclonal antibody specific for the N-linked complex-type sialo-oligosaccharide in glycoproteins. We first synthesized a chimeric immunoantigen having an N-linked complex-type of oligosaccharide of glycopeptide, which was bound to a p-formylphenyl Compound and conjugated with phosphatidylethanolamine dimylistoyl using the transglycosylation activity of a microbial endoglycosidase (Endo-M) and a reductive animation reaction. This preparative method was convenient and provided a good yield. By immunizing mice with this chimeric neoglycolipid, the monoclonal antibody for the complex-type of sialo-oligosaccharide was obtained in the culture fluid of the cell line even though it was relatively unstable. The monoclonal antibody reacted with various glycoproteins having complex-type sialo-oligosaccharides, but not with those having complex-type asialo-oligosaccharides and high mannose types of oligosaccharides, or with any glycosphingolipids. One of epitopes of this monoclonal antibody seemed to be an alpha-2,6-linked sialic acid at the non-reducing end of the sialo-oligosaccharide of the glycoprotein. (C) 2008 Elsevier Inc. All rights reserved.
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