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Time-resolved immunofluorometric assay for thyroxine-binding globulin in serum.
Y K Tan1, M J Khosravi, E P Diamandis
1CyberFluor Inc., Toronto, Ontario, Canada.
Journal of Immunoassay
|January 1, 1989
Summary
A novel immunofluorometric assay for thyroxine-binding globulin (TBG) offers a sensitive and reliable method for measuring TBG levels in serum. This assay demonstrates excellent precision and accuracy, correlating well with existing methods.
Area of Science:
- Biochemistry
- Immunology
- Analytical Chemistry
Background:
- Thyroxine-binding globulin (TBG) is crucial for thyroid hormone transport.
- Accurate measurement of TBG is essential for diagnosing thyroid disorders.
Purpose of the Study:
- To develop and validate a new "sandwich"-type immunofluorometric assay for serum TBG quantification.
- To assess the assay's performance characteristics, including sensitivity, precision, and accuracy.
Main Methods:
- Utilized a solid-phase monoclonal antibody and a biotinylated monoclonal antibody for TBG capture.
- Employed a time-resolved fluorescence detection system with a europium chelate for signal amplification.
- The assay involved a 151-fold sample pre-dilution and 90 minutes of incubation.
Main Results:
- The assay exhibited a broad dynamic range (0-100 mg/L) and a low detection limit (0.4 mg/L).
- Intra-assay and inter-assay coefficients of variation were 4.5% and 5.4%, respectively.
- Mean analytical recovery was 103%, with high correlation to radioimmunoassay (r=0.96) and immunoradiometric methods (r=0.95).
Conclusions:
- The developed immunofluorometric assay provides a precise, accurate, and sensitive method for serum TBG measurement.
- This assay is a viable alternative to existing methods for clinical diagnostics and research.