Detection of specific antigen within circulating immune complexes: validation of the assay and its application to food antigen-antibody complexes formed in healthy and food-allergic subjects

Abstract

A simple two-step method for the detection of specific antigen within immune complexes is described. The immune complexes are precipitated from serum by polyethylene glycol, dissociated by incubation in acid pH buffer and adsorbed onto the surface of polystyrene tubes. The antigen is detected by the binding of a radiolabelled affinity-purified specific antibody. The assay can detect the antigen within both antigen- and antibody-excess immune complexes of any immunoglobulin class, and can also allow semiquantitative comparison of different samples. Immune complexes containing food protein antigens after eating have been found in the serum of both normal subjects and atopic patients; the latter group showed higher mean levels of antigen-specific immune complexes. The method can be adopted for large-scale screening of clinical samples for suspected antigens if suitable antisera are available.