Functional importance of amino acid residues making up peptide antigenic determinants

Clemencia Pinilla, Jon R. Appel, Richard A. Houghten

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38 Scopus citations


The functional importance of each amino acid residue making up the antigenic determinants of three different peptide-mAb interactions was determined using complete series of substitution analogs of the three immunizing synthetic peptides. Fingerprint substitution profiles for the three different antigenic determinants were obtained separately by direct and competitive ELISA. Competitive ELISA was found to offer the advantage of being able to measure the concn of each peptide substitution analog necessary to inhibit antibody binding to the original peptide. In this manner, the relative functional contribution to antibody binding of each amino acid residue making up the antigenic determinant was determined and termed the relative positional importance factor (RPIF). Each antigenic determinant was found to contain one very highly specific residue (i.e., highest RPIF) that was, on average, the least replaceable with any of the natural l-amino acids (the average decrease in recognition ranged 250- to 28, 000-fold). At the other extreme, two or three positions in each antigenic determinant were found to be only weakly involved in recognition. These positions were considered redundant since the average decrease in recognition of the substitution analogs for these positions was found to be 20-fold or less. The remaining antigenic determinant residues exhibited the fine specificity common to antigen-antibody interactions in that only relatively conservative substitutions for these residues were recognized by their respective antibodies. It is of interest that the positional arrangement of specific and nonspecific residues were different for each of the three antigenic determinants examined.

Original languageEnglish (US)
Pages (from-to)577-585
Number of pages9
JournalMolecular Immunology
Issue number6
StatePublished - Apr 1993
Externally publishedYes


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