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Clinical Chemistry 33: 2281-2283, 1987;
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Clinical Chemistry, Vol 33, 2281-2283, Copyright © 1987 by American Association for Clinical Chemistry

Double-label time-resolved immunofluorometry of lutropin and follitropin in serum

I Hemmila, S Holttinen, K Pettersson and T Lovgren
Wallac Biochemical Laboratory, Turku, Finland.

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.


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Copyright © 1987 by the American Association for Clinical Chemistry.