title: Ultrasensitive enzyme-linked immunosorbent assay (ELISA) for the detection of picogram quantities of IgE.
abstract: Existing methods of measuring IgE in in vitro peripheral blood lymphocyte (PBL) cultures are not sufficiently sensitive to detect IgE when it is present in small amounts. This paper describes a modification of a two-site ELISA which increases the sensitivity of the assay 10-20-fold. By using the Fab' fragment of either rabbit or mouse monoclonal anti-IgE conjugated to alkaline phosphatase (AP) as the detector, the background of the assay was reduced sufficiently to permit signal amplification, using a commercially available amplified AP substrate. With this assay as little as 10 pg/ml of IgE could be detected. The interassay coefficient of variation was 15-18% between 1200 and 100 pg/ml IgE (n = 14) and there was a good correlation with a commercial IgE radioimmunoassay (RIA) (r = 0.98, n = 38).
authors.affiliation: Department of Allergy and Allied Respiratory Disorders, United Medical School, Guy's Hospital, London, U.K.
authors.forename: D M
authors.initials: DM
authors.lastname: Kemeny
authors.forename: D
authors.initials: D
authors.lastname: Richards
authors.forename: S
authors.initials: S
authors.lastname: Durnin
authors.forename: A
authors.initials: A
authors.lastname: Johannsson
mesh_terms: D000469:Alkaline Phosphatase; D000911:Antibodies, Monoclonal / Q000276:immunology; D003877:Dermatitis, Contact / Q000276:immunology; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D007073:Immunoglobulin E / Q000032:analysis*
chemical_list: D000911:Antibodies, Monoclonal; D007073:Immunoglobulin E; D000469:Alkaline Phosphatase