Epitope specificity and immunoaffinity purification of the major peanut allergen, Ara h I.
The antigenic and allergenic structure of Ara h I, a major allergen of peanuts, was investigated with the use of seven monoclonal antibodies obtained from BALB/c mice immunized with purified Ara h I. Previous work with monoclonal antibodies produced to allergens has primarily been done with inhalant allergens. Only recently have the major allergens of various foods been determined so that investigations with monoclonal antibodies into the allergenic epitopes could begin. When used as a solid phase in an ELISA, these monoclonal antibodies captured peanut antigen, which bound human IgE from patients with positive results to challenges to peanuts. The Ara h I monoclonal antibodies were found to be specific for peanut antigens when binding for other legumes was examined. In ELISA inhibition studies with the monoclonal antibodies, we identified four different antigenic sites on Ara h I. In related studies with pooled human IgE serum from patients with positive results to challenges to peanuts, we identified three similar IgE-binding epitopes. As a means of purifying the Ara h I allergen, we prepared an immunoaffinity column with monoclonal antibody 8D9. We eluted from this column the allergen Ara h I, which had a mean molecular weight of 63.5 kd and which bound human IgE from individual and pooled serum of patients with peanut sensitivity.
Burks
A W
AW
Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock.
Cockrell
G
G
Connaughton
C
C
Helm
R M
RM
D000485:Allergens / Q000276:immunology* / Q000302:isolation & purification; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D000941:Antigens / Q000276:immunology; D010367:Arachis / Q000276:immunology*; D001666:Binding Sites, Antibody; D002675:Child, Preschool; D002846:Chromatography, Affinity; D003876:Dermatitis, Atopic / Q000209:etiology / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D005260:Female; D005512:Food Hypersensitivity / Q000150:complications / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight
D000485:Allergens; D000911:Antibodies, Monoclonal; D000941:Antigens; D000939:Epitopes; D007073:Immunoglobulin E