Fluorescence sandwich enzyme-linked immunosorbent assay for detecting human interleukin-2 receptors.
A very sensitive fluorescence sandwich enzyme-linked immunosorbent assay (FS-ELISA) for the detection of soluble and cell-associated human interleukin-2 receptors (IL-2R) has been developed. For use in this assay system, two anti-human IL-2R monoclonal antibodies, H 48 and biotinylated HA 26, were selected from six monoclonal antibodies directed against different epitopes on the IL-2R molecule. The FS-ELISA specifically detected human IL-2R in both the culture supernatants and cell lysates of human IL-2R-bearing cells and the assay displayed a 10(3)-fold increase in sensitivity over the usual colorimetric IL-2R ELISA. The IL-2R molecules in supernatants and lysates of the PHA-stimulated mononuclear cells were of 50,000 and 60,000 molecular weight, respectively, as estimated by size exclusion high performance liquid chromatography.
D000911:Antibodies, Monoclonal / Q000276:immunology; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000032:analysis; D008213:Lymphocyte Activation; D008970:Molecular Weight; D011971:Receptors, Immunologic / Q000032:analysis* / Q000276:immunology; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D003470:Culture Media; D000939:Epitopes; D011971:Receptors, Immunologic; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins