Preparation and characterization of monoclonal antibodies to an N-linked oligosaccharide
| Autor: | Nobuko Miyazawa, Atsushi Nishikawa, Shigeru Fujii, Hirokazu Matsukawa, Naoyuki Taniguchi, Seizo Masutani, Takashi Shimano, Takesada Mori |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
medicine.drug_class
Blotting Western Biophysics Carbohydrates Asialoglycoproteins Biotin Oligosaccharides Chitobiose Monoclonal antibody Biochemistry Epitope Immunoglobulin G Antigen-Antibody Reactions chemistry.chemical_compound Antibody Specificity medicine Humans Molecular Biology chemistry.chemical_classification Hybridomas biology Hydrolysis Antibodies Monoclonal Cell Biology Oligosaccharide Molecular biology chemistry Polyclonal antibodies biology.protein Indicators and Reagents Antibody Glycoprotein Glucosidases |
| Zdroj: | Analytical biochemistry. 188(1) |
| ISSN: | 0003-2697 |
| Popis: | Two monoclonal antibodies to an N-linked oligosaccharide, MT-5 and MT-9, have been prepared by immunization with a pyridylaminated, asialylated, galactosylated, fucosylated, bisected biantennary sugar. The reactivity of these antibodies was monitored by their reaction with human asialoglycophorin in a solid-phase enzyme-linked immunosorbent assay. Both antibodies reacted with the sugar chains of various human glycoproteins such as immunoglobulin G, transferrin, gamma-glutamyl transpeptidase, alpha 1-acid glycoprotein, and alpha-fetoprotein. Treatment of asialoglycophorin with beta-N-acetylhexosaminidase or alpha-mannosidase resulted in reduction of the binding to these antibodies. The reactivity of MT-5 to asialoglycophorin was slightly inhibited by D-mannose and N-acetylglucosamine, whereas that of MT-9 was inhibited by D-mannose, N-acetyl-D-glucosamine, chitobiose, and L-fucose. The epitope specificity of MT-5 appears to be a sugar chain containing biantennary N-acetyl-D-glucosamine residues, the bisected N-acetyl-D-glucosamine residue, and a trimannosyl core. The epitope to which MT-9 is directed may be a complex made up of beta-mannose, chitobiose, and L-fucose. These studies indicate that immunization with pyridylaminated sugars can produce antibodies that recognize N-linked oligosaccharides. Monoclonal/polyclonal antibodies to the N-linked sugar chains of glycopeptides would be useful in such studies of proteins. |
| Databáze: | OpenAIRE |
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