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Related Experiment Videos

Recombinant human TSH receptor expressed in E. coli

A Nakai1, Y Sawai, K Miura

  • 1Department of Internal Medicine, Fujita Health University School of Medicine, Aichi, Japan.

Clinica Chimica Acta; International Journal of Clinical Chemistry
|July 4, 1997
PubMed
Summary

Researchers developed recombinant human TSH receptor (TSHR) proteins to detect TSHR autoantibodies (TRAb). These antigens can differentiate between thyroid stimulating antibodies (TSAb) and blocking antibodies (TSBAb) in autoimmune thyroid disease.

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Area of Science:

  • Endocrinology
  • Immunology
  • Molecular Biology

Background:

  • Autoimmune thyroid diseases (AITDs) involve autoantibodies targeting the TSH receptor (TSHR).
  • Distinguishing between thyroid stimulating antibodies (TSAb) and thyroid stimulating blocking antibodies (TSBAb) is crucial for diagnosis and management.
  • Current diagnostic methods for TSHR autoantibodies (TRAb) can be complex and require specific reagents.

Purpose of the Study:

  • To express recombinant extracellular domains of the human TSHR as potential antigens.
  • To develop a method for detecting TRAb and differentiating TSAb from TSBAb.
  • To evaluate the utility of these recombinant proteins in diagnosing AITDs.

Main Methods:

  • Expressed the full extracellular domain (20-408 aa) and two sub-domains (20-218 aa and 217-408 aa) of human TSHR using E. coli and pGEX.3X.

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  • Utilized Western blotting to analyze sera from patients with Graves' disease, idiopathic myxedema, and healthy controls.
  • Assessed antibody reactivity against the full-length and truncated TSHR fusion proteins.

    • Main Results:

    • The full-length TSHR fusion protein (T) reacted with sera from patients with AITDs (Graves' and idiopathic myxedema) but not with control sera.
    • Graves' disease sera (positive for TSAb) reacted with both TSHR sub-domains (5' and 3').
    • Idiopathic myxedema sera (positive for TSBAb) did not react with either TSHR sub-domain.

    Conclusions:

    • Recombinant TSHR extracellular domain proteins can serve as effective antigens for detecting TRAb.
    • The differential reactivity of TSHR sub-domains allows for the distinction between TSAb and TSBAb.
    • These recombinant antigens offer a promising tool for improved diagnosis and characterization of autoimmune thyroid diseases.