Discrimination of epitopes identified by monoclonal antibodies by competitive binding to nitrocellulose bound antigens.
A new method is described for determining the distribution of epitopes identified by monoclonal antibodies. The method utilizes nitrocellulose membranes as a solid support for antigens which are rapidly adsorbed to nitrocellulose by vacuum-blotting and then used in competitive antibody binding assays. The distribution of epitopes is established by the reciprocal cross-blocking of radiolabeled antibody by increasing concentrations of unlabeled antibody. When unlabeled antibody does not block the binding of labeled antibody to antigen, the 2 antibodies recognize distinct epitopes. When unlabeled antibody blocks the binding of labeled antibody to antigen, the 2 antibodies recognize the same epitope. The method is rapid, sensitive and should be applicable to screening monoclonal antibodies to any epitope.
Freeman
J W
JW
Chatterjee
A
A
Busch
H
H
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001667:Binding, Competitive; D002466:Cell Nucleolus / Q000276:immunology; D003101:Collodion; D000939:Epitopes; D006367:HeLa Cells; D006801:Humans; D010750:Phosphoproteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010750:Phosphoproteins; D003101:Collodion