[Food allergy: identifying and characterizing peanut allergens with patient sera and monoclonal antibodies].
The purpose of this study is to improve the diagnosis of peanut allergy. In order to standardize test substances for in vivo and in vitro diagnostic, the type I allergy-associated single components of peanuts have been identified and characterized with the aid of patients' sera and monoclonal antibodies. For allergen detection IEF-immunoprint-, SDS-PAGE-immunoblot- and 2-D electrophoresis-techniques have been used. A comparison of control sera and patients' sera showed that both contained peanut specific IgG-, IgA- and IgM-antibodies. In contrast, peanut-specific IgE-antibodies were only detectable with patients' sera. In IEF-immunoprint the most intensive IgE-bindings showed up in pl-range from pH 5.5 to 7.5. In SDS-PAGE-immunoblot major allergens could be identified at molecular weight ranges of 17, 30, 48 to 66 and 116 kD. Raising monoclonal antibodies against IgE-reactive components from peanut extract resulted in eight antibody-producing hybridoma cell lines, named PN-a to PN-h. ELISA-inhibition tests revealed common epitopes of monoclonal antibodies and patients' antibodies. Moreover, the monoclonal antibodies were tested to see whether they can be used for detection of hidden peanut allergens.
Uhlemann
L
L
Forschungsinstitut Borstel, FRG.
Becker
W M
WM
Schlaak
M
M
D000485:Allergens / Q000032:analysis* / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D010367:Arachis / Q000009:adverse effects* / Q000276:immunology; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005512:Food Hypersensitivity / Q000175:diagnosis*; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology*; D007136:Immunoglobulins / Q000097:blood / Q000276:immunology; D007525:Isoelectric Focusing; D051379:Mice; D008807:Mice, Inbred BALB C
D000485:Allergens; D000911:Antibodies, Monoclonal; D007106:Immune Sera; D007136:Immunoglobulins; D007073:Immunoglobulin E