title: A four-step sandwich radioimmunoassay for direct selection of monoclonal antibodies to allergen molecules.
abstract: A 4-step radioimmunoassay has been devised for direct identification of monoclonal antibodies (MAb) directed to IgE-binding molecules. Polyvinyl chloride wells coated with purified anti-mouse kappa chain MAb (187-1) were successively incubated with: (1) MAb-containing hybridoma supernatants, (2) allergen extract. (3) allergic patients' serum pool, and (4) 125I-labeled anti-human IgE antiserum, to detect MAb-allergen-IgE complexes. MAb to allergens from Parietaria judaica pollen and Dermatophagoides mites have been selected with this screening procedure. The affinity-purified allergen molecules completed the binding of IgE to allergen extracts coated to paper discs in a RAST inhibition assay, confirming the anti-allergen specificity of the selected MAb. This screening method is sensitive enough to allow detection of MAb directed to poorly represented allergens.
mesh_terms: D000485:Allergens / Q000276:immunology*; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D000918:Antibody Specificity; D002846:Chromatography, Affinity; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007073:Immunoglobulin E / Q000276:immunology* / Q000302:isolation & purification; D007145:Immunoglobulin kappa-Chains / Q000276:immunology; D051379:Mice; D008925:Mites / Q000276:immunology; D010944:Plants / Q000276:immunology; D011852:Radioallergosorbent Test; D011863:Radioimmunoassay
chemical_list: D000485:Allergens; D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D007145:Immunoglobulin kappa-Chains; C026408:anti-IgE antibodies; D007073:Immunoglobulin E