Comparison of different methods for localizing antigenic regions in histone H2A

Abstract

Four different variations of enzyme-linked immunosorbent assays (ELISA) were used to analyze the antigenic structure of histone H2A. Eleven natural and 10 synthetic peptides of H2A were tested for their capacity to bind antibodies raised against the complete molecule in a direct binding assay. Results were compared to those obtained in a direct test using several peptide-BSA conjugates. The capacity of peptides to inhibit the reaction between H2A antibodies and the complete H2A molecule or large fragments of it was also measured. Inhibition assays detected antigenic activity in a large number of peptides than did direct binding assays. Antisera raised against eight synthetic, unconjugated peptides all reacted with histone H2A in ELISA. Using as probes peptides of 14-21 residues, at least 11 antigenic regions could be recognized, indicating that virtually the entire H2A polypeptide chain possessed antigenic activity.