Detection of house dust mite allergens and immunoblot analysis in asthmatic children

The mite allergen extracts (Dermatophagoides pteronyssinus and D. farinae), were fractionated by polyacrylamide gel electrophoresis and autoradiography. The molecular weight of the most apparent proteins of D. pteronyssinus was between 27,000 and 35,000, and the protein components of D. farinae were in the 9-100-kilodalton range. Specific IgE antibodies directed against mite allergens were utilized as probes to detect the allergens. Using immunoblot analysis and radiostained technique, D. pteronyssinus-sensitive patient sera displayed the major protein of allergens with molecular weights of 15,800 and 27,000-31,000, while D. farinae-sensitive patients' sera displayed the major protein of allergens with molecular weights of 12,000, 15,800 and 26,000-29,000. The electroblotting technique is fast, convenient, and highly suitable for both allergen composition studies and screening of antibody specificities.