Abstract
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.
Original language | English (US) |
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Pages (from-to) | 259-265 |
Number of pages | 7 |
Journal | Journal of Immunological Methods |
Volume | 78 |
Issue number | 2 |
DOIs | |
State | Published - Apr 22 1985 |
Keywords
- epitopes
- immunoassay
- monoclonal antibody
- nitrocellulose binding
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology