Novel methods to determine the epitopes on the asparagine-linked oligosaccharides of glycoproteins.
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.
Ohbayashi
H
H
Department of Biochemistry, University of Tokyo, Japan.
Endo
T
T
Yamashita
K
K
Kuroki
M
M
Matsuoka
Y
Y
Kobata
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001215:Asparaginase; D001216:Asparagine; D002417:Cattle; D002855:Chromatography, Thin Layer; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D051379:Mice; D008832:Microchemistry; D009844:Oligosaccharides / Q000276:immunology*; D011107:Polylysine / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006023:Glycoproteins; D009844:Oligosaccharides; D011107:Polylysine; D001216:Asparagine; D001215:Asparaginase