title: Conformational differences of human IgE on hydrophobic and hydrophilic solid phases detected by monoclonal antibodies.
abstract: Very sensitive assays of IgE are required for determining prevalence of allergic reactions in children. In order to develop a sensitive two-site IRMA two kinds of mAb were produced. Antibodies specific for D epsilon 1 determinants were derived from immunization with a 40 kDa papain Fc fragment. They bound equally native and 56 degrees C heated IgE. D epsilon 2 specific mAb were obtained after immunization with IgE anti-D epsilon 1 complex and were selected on the basis of their inability to bind heated IgE. In a two-site assay on plastic plates, D epsilon 1 specific mAb led to the binding of IgE but always prevented further binding of anti-D epsilon 1 mAb, anti-human kappa chain mAb or allergen on bound IgE. This was not true when CNBr activated cellulose was used. The influence of the nature of the solid phase disappeared when D epsilon 2 specific mAb were coated on plastic tubes. In this case, the binding of a second mAb with identical or different fine specificity was observed. The best matched pair was E 164 (anti-D epsilon 2) on the solid phase and 6H10 (anti-D epsilon 1) as a tracer. As little as 0.2 UI/ml of IgE could be detected in a 2 hr test.
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000939:Epitopes / Q000276:immunology; D005260:Female; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis* / Q000276:immunology; D007141:Immunoglobulin Fc Fragments / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010969:Plastics; D011487:Protein Conformation; D011863:Radioimmunoassay
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D007141:Immunoglobulin Fc Fragments; D010969:Plastics; D007073:Immunoglobulin E