Discrimination of epitopes identified by monoclonal antibodies by competitive binding to nitrocellulose bound antigens

James W. Freeman, Amitava Chatterjee, Harris Busch

Research output: Contribution to journalArticle

8 Scopus citations

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 languageEnglish (US)
Pages (from-to)259-265
Number of pages7
JournalJournal of Immunological Methods
Volume78
Issue number2
DOIs
StatePublished - Apr 22 1985

Keywords

  • epitopes
  • immunoassay
  • monoclonal antibody
  • nitrocellulose binding

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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