Double-label time-resolved immunofluorometry of lutropin and follitropin in serum.
We describe a procedure for the simultaneous immunofluorometric assay of lutropin and follitropin in human serum, based on the use of monoclonal antibodies and of the fluorescent lanthanides Eu3+ and Tb3+. The alpha-chain-specific antibody was used as a common capture antibody on the surfaces of microtitration strips. The anti-beta-follitropin antibody was labeled with Tb3+, the anti-beta-lutropin antibody with Eu3+. After the immunoreactions had taken place, the bound fractions of the labels were dissociated in a fluorescence enhancement solution of pivaloyltrifluoroacetone, trioctylphosphine oxide, and Triton X-100 surfactant. In this solution both lanthanides can be measured successively with a time-resolved fluorometer. The detection limit of the assay is 0.1 int. unit/L for lutropin and 1 int. unit/L for follitropin. Results correlated well with those by commercial immunofluorometric assays and radioimmunoassays.
D000911:Antibodies, Monoclonal; D005063:Europium; D005470:Fluorometry; D005640:Follicle Stimulating Hormone / Q000097:blood*; D006801:Humans; D007118:Immunoassay; D007986:Luteinizing Hormone / Q000097:blood*; D013725:Terbium
D000911:Antibodies, Monoclonal; D013725:Terbium; D005063:Europium; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone