Highly sensitive-beta-D-galactosidase-linked immunosorbent assay for anti-myelin basic protein antibody.
A highly sensitive enzyme-linked immunosorbent assay (ELISA) procedure has been devised for the determination of anti-myelin basic protein antibodies. First, rabbit anti-myelin basic protein (MBP) sera to be assayed were incubated with MBP immobilized on small pieces of silicone rubber. Next, anti-MBP bound specifically to the solid phase was allowed to react either with Fab' of guinea pig anti-rabbit IgG conjugated with beta-D-galactosidase from Escherichia coli or with protein A conjugated with beta-D-galactosidase. The amount of fixed anti-MBP antibodies was assayed by fluorometric determination of the enzyme activity using 4-methylumbelliferyl-beta-D-galactoside as a substrate. The sensitivity of the present assay was at least an order of magnitude higher than the hitherto available radioimmunoassay and ELISA.
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005696:Galactosidases; D006168:Guinea Pigs; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D004676:Myelin Basic Protein / Q000276:immunology*; D001616:beta-Galactosidase
D000906:Antibodies; D004676:Myelin Basic Protein; D005696:Galactosidases; D001616:beta-Galactosidase