title: Microparticle-enhanced nephelometric immunoassay of anti-thyroid peroxidase autoantibodies in thyroid disorders.
abstract: Crude thyroid peroxidase extracted from human thyroid microsomes was covalently bound onto polyacrylic and polyfunctional copolymerized microparticles. We observed agglutination of the thyroid peroxidase-microparticle conjugate with 13 monoclonal antibodies (mAbs) specific for epitopes on four different antigenic domains of human thyroid peroxidase (TPO; EC 1.11.1.7), after addition of anti-mouse immunoglobulins. We quantified agglutination by measuring with a specially designed nephelometer the light scattered by the conjugates. This allowed us to develop a microparticle-enhanced nephelometric immunoassay for human anti-TPO autoantibodies (aAbs) with defined epitopic specificity, based on the ability of aAbs to inhibit mAb-induced agglutination. Applied to patients with autoimmune thyroid diseases, this assay confirmed the polyclonality of anti-TPO aAbs and their preferential reactivity toward epitopes located on the A and B antigenic domains of the TPO molecule. The same specificities seem to be present in patients with Hashimoto thyroiditis or Graves disease.
authors.affiliation: Immunology Laboratory, Faculty of Medicine, Vandoeuvre, France.
authors.forename: A A
authors.initials: AA
authors.lastname: Harchali
authors.forename: P
authors.initials: P
authors.lastname: Montagne
authors.forename: J
authors.initials: J
authors.lastname: Ruf
authors.forename: M L
authors.initials: ML
authors.lastname: Cuillière
authors.forename: M C
authors.initials: MC
authors.lastname: Bene
authors.forename: G
authors.initials: G
authors.lastname: Faure
authors.forename: J
authors.initials: J
authors.lastname: Duheille
mesh_terms: D001323:Autoantibodies / Q000097:blood*; D000939:Epitopes / Q000276:immunology; D006111:Graves Disease / Q000276:immunology; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007453:Iodide Peroxidase / Q000276:immunology*; D008863:Microspheres; D009391:Nephelometry and Turbidimetry; D013959:Thyroid Diseases / Q000276:immunology*; D013967:Thyroiditis, Autoimmune / Q000276:immunology
chemical_list: D001323:Autoantibodies; D000939:Epitopes; C056626:thyroid microsomal antibodies; D007453:Iodide Peroxidase