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

• Published in: Journal of allergy and clinical immunology Vol. 107; no. 5; pp. 907 - 913
• Main Authors: Sander, Ingrid, Flagge, Angelika, Merget, Rolf, Halder, Thomas M., Meyer, Helmut E., Baur, Xaver
• Format: Journal Article Conference Proceeding
• Language: English
• Published: New York, NY Mosby, Inc 01.05.2001; Elsevier
• Subjects: Allergens - adverse effects; Allergens - immunology; Allergens - isolation & purification; Allergic diseases; Antibody Specificity; Asthma - blood; Asthma - etiology; Asthma - immunology; Baker’s asthma; Biological and medical sciences; Blotting, Western; Cooking; Electrophoresis, Gel, Two-Dimensional; Flour - adverse effects; Flour - analysis; glyceraldehyde-3-phosphate dehydrogenase; Glyceraldehyde-3-Phosphate Dehydrogenases - adverse effects; Glyceraldehyde-3-Phosphate Dehydrogenases - immunology; Glyceraldehyde-3-Phosphate Dehydrogenases - isolation & purification; glycerinaldehyde-3-phosphate dehydrogenase; Hordeum - enzymology; Hordeum vulgare; Humans; Hydrogen-Ion Concentration; Immunoglobulin E - blood; Immunoglobulin E - immunology; immunoglobulin E-binding protein; Immunopathology; Medical sciences; Occupational Diseases - blood; Occupational Diseases - etiology; Occupational Diseases - immunology; Other localizations; Plant Proteins - adverse effects; Plant Proteins - immunology; Plant Proteins - isolation & purification; Protein Isoforms - adverse effects; Protein Isoforms - immunology; Protein Isoforms - isolation & purification; serine proteinase inhibitors; serpin; serpins; Serpins - adverse effects; Serpins - immunology; Serpins - isolation & purification; Solubility; Species Specificity; Triose-Phosphate Isomerase - adverse effects; Triose-Phosphate Isomerase - immunology; Triose-Phosphate Isomerase - isolation & purification; triosephosphate isomerase; Triticum - adverse effects; Triticum - chemistry; Triticum - immunology; Triticum aestivum; Water; wheat flour allergens; wheat flour allergens; IPG; TBS; DTT; Baker’s asthma; serpin; glycerinaldehyde-3-phosphate dehydrogenase; pI; triosephosphate isomerase; EAST; PVDF; CHAPS; Allergy; Monocotyledones; Intramolecular oxidoreductases; Pathogenesis; IgE; Occupational exposure; Triose-phosphate isomerase; Glyceraldehyde-3-phosphate dehydrogenase (phosphorylating); Isomerases; Gramineae; Immunological investigation; Baker; Angiospermae; Immunoblotting assay; Obstructive pulmonary disease; Human; Immunopathology; Hordeum vulgare; Specific IgE; Respiratory disease; Enzyme; Asthma; Serpin; Wheat flour; Spermatophyta; Oxidoreductases; Triticum aestivum; Allergen
• ISSN: 0091-6749
• DOI: 10.1067/mai.2001.113761

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.)