Analysis of antibodies that recognize the B-epitopes formed on protein antigens by glutaraldehyde treatment: an...

Immunochemical analysis of antigenic determinants (AD) formed on protein macromolecules by glutaraldehyde (GA) treatment with subsequent block of free aldehyde groups with glycine was performed. The structure of the epitopes was analyzed using BSA and ovalbumin (Ova) treated with GA with subsequent block of the active groups with glycine and other amino acids. The products of reaction of GA with Gly, Lys, Pro, and His were used as hapten-like antigens that mimic GA antigenic determinants (GA-AD). Indirect and competitive enzyme-linked immunoassay and immunoaffinity chromatography revealed two regions of epitope density in the structure of the GA determinant. It is shown that the product of GA reaction with Gly efficiently blocks the antibodies raised against GA-treated proteins. Based on this finding, a method of assay of the peptide-specific antibodies of the immune complexes with the peptide--protein conjugate in the presence of antibodies to the linking agents is suggested.