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Related Experiment Videos

A quick method to detect circulating anti-thyroid hormone autoantibodies

S Savastano1, A P Tommaselli, R Valentino

  • 1Cattedra di Endocrinologia, Facoltà di Medicina e Chirurgia, Università Federico II, Napoli, Italy.

Journal of Endocrinological Investigation
|January 1, 1995
PubMed
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This study introduces a new chromatographic method to detect anti-thyroid hormone autoantibodies (THBA) and measure thyroid hormone (TH) levels. The method accurately identifies THBA, aiding in diagnosing thyroid conditions with potentially misleading hormone readings.

Area of Science:

  • Endocrinology
  • Clinical Chemistry
  • Analytical Chemistry

Background:

  • Circulating anti-thyroid hormone autoantibodies (THBA) can interfere with accurate thyroid hormone (TH) measurements.
  • Existing methods for THBA detection and TH quantification in their presence may be complex or time-consuming.
  • Accurate assessment of TH levels is crucial for diagnosing and managing thyroid disorders.

Purpose of the Study:

  • To develop and validate a straightforward chromatographic method for detecting THBA.
  • To determine the affinity constant of THBA.
  • To accurately quantify total thyroid hormone (TH) levels in samples containing THBA.

Main Methods:

  • A chromatographic technique using Sephadex LH 20 columns was employed.
  • Samples were incubated with radiolabeled T3 or T4 tracers and an inhibitor to prevent plasma protein binding.

Related Experiment Videos

  • Separation of free TH from THBA-bound TH was achieved using TRIS buffer elution, followed by methanol elution for THBA-bound TH quantification.

    • Main Results:

    • The method successfully distinguished THBA-positive samples from controls based on radioactivity levels.
    • THBA-bound TH were eluted and quantified, allowing for the calculation of total TH levels.
    • Validation against two standardized methods confirmed the reliability of the proposed assay.

    Conclusions:

    • The developed chromatographic method is a feasible and efficient tool for detecting THBA.
    • This assay enables accurate measurement of total TH levels even in the presence of THBA.
    • The method offers a rapid solution for samples with suspected spuriously high free and total TH values, aiding in thyroid diagnostics.