Crossed Immunoelectrophoretic and Crossed Radioimmunoelectrophoretic Studies Employing a Model Allergen from Codfish

Abstract

A standard reference pattern in crossed immunoelectrophoresis (CIE) of the parvalbumin fraction of a cod white muscle extract was established. Crossed line immunoelectrophoresis (CLIE), tandem CIE and crossed radioimmunoelectrophoresis (CRIE) were used to identify the major allergens, thereby establishing a basis for future work employing quantitative immunoelectrophoretic techniques in the study of codfish allergens. Rocket immunoelectrophoresis and rocket-line immunoelectrophoresis were used to demonstrate antigenic determinants on polypeptide fragments of the major allergen, Allergen M. The results show the usefulness of these techniques in studies of antigenic determinants on non-precipitating polypeptides. CRIE with 8 patient’s sera showed radiostaining indicating IgE binding corresponding to at least 7 precipitates in the crude extract CIE preparations. The purified DS 22 fraction was shown to contain 2 CRIE-positive (IgE-binding) precipitates. When using Allergen M, these precipitates were also demonstrated, one of them in trace amounts only.