Antibody binding of deletion mutants of Asp f 2, the major Aspergillus fumigatus allergen. Biochem Biophys Res Commun 2000 Apr 21;270(3):1128-35
Date
04/25/2000Pubmed ID
10772962DOI
10.1006/bbrc.2000.2546Scopus ID
2-s2.0-0034696795 (requires institutional sign-in at Scopus site) 24 CitationsAbstract
Asp f 2, a 268 amino acid major allergen from Aspergillus fumigatus (Af) demonstrated nine linear IgE binding regions. It is not known whether any of these linear epitopes are also conformatory epitopes. Hence, we constructed deletion mutants of Asp f 2 devoid of one or more epitopes, and the IgE binding of these proteins with sera from patients with ABPA was compared with the full-length Asp f 2 expressed in E. coli and Pichia. The Pichia expressed protein reacted weakly with IgE, but strongly with IgG of ABPA sera compared to E. coli expressed Asp f 2. Weak IgE binding only was seen when the C-terminal or N-terminal was deleted, while depletion of both ends negated all reactivity. The monoclonal antibody IL-B8 and IgE and IgG of ABPA sera bound significantly to the Asp f 2 E-4 fragment indicating that the major B-cell epitope is located at the N-terminal end of Asp f 2.
Author List
Tang B, Banerjee B, Greenberger PA, Fink JN, Kelly KJ, Kurup VPMESH terms used to index this publication - Major topics in bold
AllergensAmino Acid Sequence
Aspergillosis, Allergic Bronchopulmonary
Aspergillus fumigatus
Binding Sites, Antibody
Cloning, Molecular
Escherichia coli
Fungal Proteins
Humans
Immunoglobulin E
Immunoglobulin G
Molecular Sequence Data
Pichia
Recombinant Proteins
Reference Values
Sequence Deletion
