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Monoclonal antibodies to three distinct epitopes on human IgE: their use for determination of allergen-specific IgE
Authors:F Sánchez-Madrid  G Morago  A L Corbi  J Carreira
Affiliation:Departamento de Investigación, Alergia e Immunología Abelló, Julián Camarillo, 8 Madrid, 17 Spain
Abstract:Three different monoclonal antibodies (MAb) against human immunoglobulin E have been obtained which specifically bind to human myeloma and polyclonal IgE. The antibodies showed high avidities for soluble IgE (0.7 X 10(9) to 3.3 X 10(9) M-1). These MAb defined three distinct epitopes on IgE. A mixture of these antibodies in combination with an 125I-labelled anti-mouse Kappa chain MAb has been used to measure allergen-specific IgE. This determination was performed by a solid-phase radioimmunoassay using allergen extracts coated to either chemically activated paper discs or to polyvinyl chloride wells. This method is 4-10 times more sensitive than other previously reported procedures. A similar technique has also been applied to detect individual allergens in immunoblots of allergen extracts.
Keywords:IgE determination  monoclonal antibodies  allergen molecules  BSA  bovine serum album  DMEM  Dulbeco's modified Eagle's medium  FCS  fetal calf serum  HE  monoclonal antibody(es) to human IgE  HG  monoclonal antibody(es) to human IgG  Ig  immunoglobulin(s)  MAb  monoclonal antibody(ies)  PAGE  polacrylamide gel electrophoresis  PBS  0.15 NaCl  0.01 M sodium phosphate  pH 7.2  SDS  sodium dodecyl sulfate
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