[Detection and analysis of IgG antibodies in sera of patient with atopic dermatitis (AD) to individual wheat polypeptides by immunoblotting techniques].

To detect IgG antibodies in sera of patients with atopic dermatitis (AD) to individual wheat polypeptides, wheat flour proteins were extracted with tris-citrate buffered saline containing urea and 2-mercaptoethanol (UTC solvent), and fractionated as polypeptides of molecular weights between 12 and 100 KDa. By means of the immunoblotting techniques, specific IgG antibodies to fractionated proteins could be detected in sera from healthy individuals and AD patients. Twenty AD patients' sera with high titers of wheat specific RAST (scores of more than 3) reacted not only to five major wheat components (33-38, 40-45, 55-60, 80, 90-98 KDa), which stained intensively with Coomassie brilliant blue G-250, but also to polypeptides of 14, 16, 22, 29, 70 KDa. Twelve control sera reacted to a limited number of the five major wheat components, and levels of specific IgG antibodies increased with age. The sera of AD patients in the present study contained increased level of IgG antibodies to the five major components, and additionally to several minor components, 70 KDa, 29 KDa and much smaller proteins.