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Related Experiment Videos

Competitive solid-phase immunoassay of testosterone using time-resolved fluorescence.

E Bertoft, J U Eskola, V Näntö

    FEBS Letters
    |July 23, 1984
    PubMed
    Summary

    This study presents a new time-resolved fluorescence immunoassay for measuring testosterone in serum. The assay demonstrates high sensitivity and excellent correlation with radioimmunoassay, offering a reliable method for testosterone determination.

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    Area of Science:

    • Biochemistry
    • Analytical Chemistry
    • Immunology

    Background:

    • Accurate testosterone determination is crucial for diagnosing various endocrine disorders.
    • Existing methods like radioimmunoassay (RIA) have limitations, necessitating development of alternative assays.

    Purpose of the Study:

    • To develop and validate a competitive solid-phase immunoassay for testosterone quantification in serum samples.
    • To evaluate the performance of time-resolved fluorescence detection in this immunoassay.

    Main Methods:

    • A competitive solid-phase immunoassay utilizing testosterone conjugate-coated polystyrene microtiter strips.
    • Comparison of Europium-labelled polyclonal and monoclonal antibodies for testosterone detection.
    • Time-resolved fluorescence detection for signal measurement.

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    Main Results:

    • The immunoassay achieved a detection limit of 15 fmol testosterone per assay using a polyclonal antibody.
    • Excellent correlation was observed between the developed immunoassay and RIA (r = 0.982).
    • Both polyclonal and monoclonal antibodies showed similar behavior, with polyclonal exhibiting higher sensitivity.

    Conclusions:

    • The developed time-resolved fluorescence immunoassay is a sensitive and reliable method for serum testosterone determination.
    • This assay offers a viable alternative to RIA for clinical and research applications.
    • The use of Europium-labelled antibodies provides a robust platform for fluorescence immunoassays.