Purification of a major allergen, Asp f 2 binding to IgE in allergic bronchopulmonary aspergillosis, from culture flitrate of Aspergillus fumigatus

B. Banerjee, V. P. Kurup*, P. A. Greenberger, D. R. Hoffman, D. S. Nair, J. N. Fink

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

47 Scopus citations

Abstract

Background: Most cases of allergic bronchopulmonary aspergillosis (ABPA) are caused by the fungus Aspergillus fumigatus. Successful treatment of this disease depends on early diagnosis with the use of well-characterized and relevant antigens/allergens of the organism. Objective: The aim of this study was to purify and characterize relevant proteins from A. fumigatus that could be used in the reliable diagnosis of ABPA. Methods: Monoclonal antibodies were raised against A. fumigatus culture filtrate antigens. A Concanavalin A nonbinding protein fraction was purified with use of one of the monoclonal antibody immunoaffinity columns. The purified protein was analyzed on sodium dodecylsulfate-polyacrylamide gel electrophoresis gel and Western blots. The sensitivity and specificity of the purified protein were evaluated by RAST and ELISA with sera from 25 patients with ABPA, from 10 with allergic asthma, and from 10 normal control subjects. Results: The 37 kD Concanavalin A nonbinding protein reacted specifically with IgE antibodies in patients with ABPA. Among the 25 patients with ABPA studied, 96% had IgE antibody against the allergen, whereas none of the subjects with allergic asthma who had positive results on the skin prick test or normal control subjects had a reaction. Both RAST and ELISA results exhibited strong correlation with IgE binding. This allergen exhibited N-terminal sequence identity to a recombinant allergen Asp f 2. Conclusions: A 37 kD protein with complete N- terminal homology to Asp f 2 is a major allergen of A. fumigatus that significantly reacts with IgE antibody in patients with ABPA, but does not elicit reaction in Aspergillus-sensitive subjects with asthma and normal control subjects.

Original languageEnglish (US)
Pages (from-to)821-827
Number of pages7
JournalJournal of Allergy and Clinical Immunology
Volume99
Issue number6 I SUPPL.
DOIs
StatePublished - Jan 1 1997

Keywords

  • Allergen
  • Allergic bronchopulmonary aspergillosis
  • Aspergillus fumigatus
  • ELISA
  • Monoclonal antibody
  • RAST

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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