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Development and basic performance verification of a rapid homogeneous bioassay for agonistic antibodies against the thyroid-stimulating hormone receptor.

Abstract
Graves' disease is a type of autoimmune hyperthyroidism caused by thyroid-stimulating antibodies (TSAb).1 The combination of a porcine thyroid cell bioassay and cyclic adenosine monophosphate (cAMP) immunoassay (TSAb-enzyme immunoassay; EIA) is a clinically approved TSAb measurement method. Due to the requirement of multiple procedures and a long assay time of 6 h in the TSAb-EIA, a simplified and rapid assay is desired. Herein, we developed a rapid homogeneous TSAb bioassay (rapid-TSAb assay) using the human embryonic kidney cell line (HEK293), engineered to express the human thyroid-stimulating hormone receptor (TSHR), along with a cAMP-dependent luminescence biosensor. The measurement consists of three steps: thawing frozen cells, blood sample addition, and luminescence detection. The procedures can be conducted within 1 h. The World Health Organization International Standard TSAb (NIBSC 08/204) stimulated the cells co-expressing TSHR and cAMP biosensor. The intra- and inter-assay coefficients of variance were < 10%. Stimulation activity using wild-type TSHR and chimeric TSHR (Mc4) almost completely correlated with the tested Graves' disease and normal samples. In the rapid-TSAb assay, the evaluation of 39 samples, including TSHR antibody-positive sera, yielded a sensitivity of 100.0% and a specificity of 90.9%, compared to the TSAb-EIA control. The rapid-TSAb assay enables simple and rapid measurement of TSAb and is promising for improving the diagnosis of autoimmune thyroid diseases.
AuthorsMotoki Hoshina, Shiomi Ojima, Atsushi Kawasaki, Kosuke Doi, Satoshi Ohta, Asuka Inoue, Hiroshi Murayama
JournalJournal of immunological methods (J Immunol Methods) Vol. 528 Pg. 113655 (May 2024) ISSN: 1872-7905 [Electronic] Netherlands
PMID38447802 (Publication Type: Journal Article)
CopyrightCopyright © 2023. Published by Elsevier B.V.
Chemical References
  • Receptors, Thyrotropin
  • Immunoglobulins, Thyroid-Stimulating
  • Receptors, G-Protein-Coupled
  • Thyrotropin
  • Autoantibodies
Topics
  • Animals
  • Swine
  • Humans
  • Receptors, Thyrotropin
  • HEK293 Cells
  • Immunoglobulins, Thyroid-Stimulating
  • Graves Disease
  • Receptors, G-Protein-Coupled
  • Thyrotropin
  • Biological Assay (methods)
  • Autoantibodies

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