Antibody binding of deletion mutants of Asp f 2, the major Aspergillus fumigatus allergen.
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.
Tang
B
B
Department of Pediatrics, Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
Banerjee
B
B
Greenberger
P A
PA
Fink
J N
JN
Kelly
K J
KJ
Kurup
V P
VP
D000485:Allergens / Q000737:chemistry / Q000276:immunology*; D000595:Amino Acid Sequence; D001229:Aspergillosis, Allergic Bronchopulmonary / Q000097:blood / Q000276:immunology*; D001232:Aspergillus fumigatus / Q000235:genetics / Q000276:immunology*; D001666:Binding Sites, Antibody; D003001:Cloning, Molecular; D004926:Escherichia coli; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood*; D007074:Immunoglobulin G / Q000097:blood; D008969:Molecular Sequence Data; D010843:Pichia; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D012016:Reference Values; D017384:Sequence Deletion
D000485:Allergens; D005656:Fungal Proteins; D007074:Immunoglobulin G; D011994:Recombinant Proteins; C106810:allergen Asp f 2, Aspergillus fumigatus; D007073:Immunoglobulin E