Monoclonal antibody GOM-2 binds to blood group B-Ley active glycolipid antigens on human gastric cancer cells,KATO-III |
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Authors: | Shinobu Sueyoshi Hitomi Nagakura Akira Kato Setsuyoshi Uetsuki Yasuo Nakayama Masakazu Adachi |
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Affiliation: | (1) Formulation Research Institute, Otsuka Pharmaceutical Company, 771-01 Tokushima, Japan;(2) Japan Immunoresearch Laboratories, 370 Takasaki, Gunma, Japan |
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Abstract: | The antigen structure of a mouse monoclonal antibody, GOM-2, established by immunization with KATO-III human gastric cancer cells, was examined. GOM-2 reactive glycolipids were prepared from KATO-III cells and treated with endoglycoceramidase. Structural studies of ten GOM-2 reactive oligosaccharides by a combination of glycosidase digestions, methylation, and affinity chromatography on anUlex europeus agglutinin I (UEA-I) column revealed that nine of them had a Y-related B-active difucosylated determinant (B-Ley) and one had a B-active determinant. Affinity chromatography of the purified and modified oligosaccharides on an immobilized GOM-2 column demonstrated that GOM-2 has a novel binding specificity; it binds tightly to the biantennary structure carrying the B-Ley determinant at the termini or the branched structure carrying the B-Ley structure at two nonreducing termini.Abbreviations UEA-I Ulex europeus agglutinin I - PNA Arachis hypogaea agglutinin - Fuc l-fucose - Gal d-galactose - Glcol glucitol - GlcNAc N-acetyl-d-glucosamine - TBS 10mm Tris-HCl, pH 7.8, containing 150mm NaCl - PBS 10mm sodium phosphate buffer, pH 7.5, containing 150mm NaCl - HPLC high performance liquid chromatography. |
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Keywords: | B-Ley active glycolipids monoclonal antibody |
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