Sensitive time-resolved immunofluorometric assay of thyrotropin in serum.
We have developed a time-resolved solid-phase immunofluorometric assay for thyrotropin (TSH). The assay is performed in white opaque microtitration wells which are coated with a monoclonal capture antibody. Serum TSH binds simultaneously to the solid phase and to a biotinylated monoclonal detection antibody. The degree of biotinylated antibody binding is quantitated with streptavidin conjugated to thyroglobulin which is heavily labelled with the Eu3+ chelator 4,7-bis [chlorosulfophenyl] -1,10-phenanthroline -2,9-dicarboxylic acid (BCPDA). The final fluorescent complex is measured on the solid phase with time-resolved fluorometry. The assay requires two incubation steps and can be completed in 5 hours. The detection limit is 0.03 milli-int. units/L. The present assay was compared with two immunoradiometric assays and gave satisfactory results.
Reichstein
E
E
CyberFluor, Inc., Toronto, Ontario, Canada.
Morton
R C
RC
Diamandis
E P
EP
D000911:Antibodies, Monoclonal; D005067:Euthyroid Sick Syndromes / Q000175:diagnosis; D005455:Fluorescent Antibody Technique; D006801:Humans; D006980:Hyperthyroidism / Q000175:diagnosis; D007037:Hypothyroidism / Q000175:diagnosis; D013960:Thyroid Function Tests / Q000379:methods; D013972:Thyrotropin / Q000097:blood*; D013997:Time Factors
D000911:Antibodies, Monoclonal; D013972:Thyrotropin