Identification of wheat flour allergens by means of 2-dimensional immunoblotting

Abstract

Background: Wheat flour proteins are allergens for 60% to 70% of bakers with workplace-related respiratory symptoms. Objective: The aim of the study was to investigate the variability of IgE antibody patterns of wheat flour–sensitized bakers and to identify the most frequently recognized allergens. Methods: Water/salt-soluble wheat flour proteins from the cultivar Bussard were separated by using 2-dimensional gel electrophoresis with immobilized pH gradients. IgE-reactive proteins were identified by means of immunoblotting with sera of 10 subjects with baker’s asthma. Mass spectrometric fingerprinting was used to identify the proteins most frequently recognized by IgE. Results: The IgE immunoblots obtained with 10 different sera exhibited a remarkable heterogeneity. Each patient showed an individual IgE-binding pattern with 4 to 50 different allergen spots. Altogether, more than 100 IgE-binding protein spots were detected. Nine of the predominant IgE-binding protein spots were identified by using mass spectrometric fingerprinting. The obtained masses matched 2 different isoforms of glycerinaldehyde-3-phosphate dehydrogenase from Hordeum vulgare , triosephosphate isomerase from H vulgare , and serpin, a serine proteinase inhibitor from Triticum aestivum . Conclusions: The results show a great interindividual variation of IgE-binding patterns of wheat flour proteins in baker’s asthma. The clinical relevance of the identified 4 new allergens will be further investigated in the near future. (J Allergy Clin Immunol 2001;107:907-13.)