Novel methods to determine the epitopes on the asparagine-linked oligosaccharides of glycoproteins |
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Authors: | H Ohbayashi T Endo K Yamashita M Kuroki Y Matsuoka A Kobata |
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Affiliation: | Department of Biochemistry, University of Tokyo, Japan. |
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Abstract: | Oligosaccharides obtained from glycoproteins by hydrazinolysis followed by N-acetylation were covalently coupled to an amino-bonded thin-layer plate or to a poly-L-lysine coating on the microtiter plate. Bound oligosaccharides can be directly detected by immunostaining or by enzyme-linked immunosorbent assay (ELISA) using a mouse monoclonal antibody. These methods are simple and require small amounts of oligosaccharides and antibodies. The methods were successfully applied to determine the epitope of F48-60, a monoclonal antibody which reacts with the N-linked sugar chains of nonspecific cross-reacting antigen 2, but not with those of carcinoembryonic antigen. The results revealed that the antibody recognizes the Gal beta 1----3GlcNAc beta 1----group. |
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