RRC ID 52607
著者 Okuda T, Fukui A.
タイトル Generation of anti-oligosaccharide antibodies that recognize mammalian glycoproteins by immunization with a novel artificial glycosphingolipid.
ジャーナル Biochem Biophys Res Commun
Abstract Here we report a new method for the efficient generation of antibodies that recognize the fine structures of oligosaccharides on glycoproteins. We found a newly designed artificial glycosphingolipid carrying a very long chain fatty acid to be a strong immunogen in mice, with the serum of immunized mice containing antibodies recognizing the oligosaccharide structure of the immunogen. First, we found that conjugation of a simple ceramide analogue to target oligosaccharides could enhance the immunogenicity of these oligosaccharides in these immunized mice. This effect was confirmed in mice immunized with the artificial glycosphingolipids carrying 6'-Sialyl-LacNAc, 3'-Sialyl-LacNAc and LacNAc. Next, we tried to improve the immunogenic enhancing effect of the ceramide analogue. In a model experiment using 6'-Sialyl-LacNAc oligosaccharide, we manipulated the alkyl chains to several lengths, and found that a longer alkyl chain length of the fatty acid correlated with high immunogenicity. Among these we examined, artificial glycosphingolipids conjugated with a ceramide analogue carrying a very long chain fatty acid (lignoceric acid) showed the strongest immunogenicity. By using the artificial glycosphingolipid containing 6'-Sialy-LacNAc and lignoceric acid, we succeeded in the generation of a new anti-6'-Sialyl-LacNAc antibody that recognizes 6'-Sialyl-LacNAc carrying glycoproteins but does not bind to 6'-Sialyllactose, asialo-glycoporoteins and glycoproteins carrying 3'-Sialyl-LacNAc. These results indicate that the established technology is valuable for the targeted generation of monoclonal antibodies against glycoproteins containing specific oligosaccharide structures.
巻・号 497(4)
ページ 983-989
公開日 2018-3-18
DOI 10.1016/j.bbrc.2018.02.113
PII S0006-291X(18)30342-5
PMID 29458022
MeSH Animals Antibodies, Monoclonal / immunology* Glycoproteins / chemistry Glycoproteins / immunology Glycosphingolipids / chemical synthesis Glycosphingolipids / chemistry Glycosphingolipids / immunology Immunization Methods Mice Oligosaccharides / immunology* Sensitivity and Specificity
IF 2.985
引用数 7
リソース情報
ヒト・動物細胞 Sp2/0-Ag14(RCB0209)