Immunoblotting for the detection of TSH receptor autoantibodies.
Immunoblotting was optimized to detect autoantibodies to TSH receptors from human and porcine thyroid tissue and to determine their epitope specificity. Autoantibodies to putative TSH receptor proteins in thyroid particulate membranes were detected in approximately 35% of sera from patients with Graves' disease. However, despite modifications to increase immunoblotting sensitivity and specificity, only a minority (less than 15%) of Graves' disease sera contained autoantibodies that identified epitopes within TSH affinity-purified human or porcine receptor proteins. In these sera there was no correlation between the TSH receptor antibody titre, determined by radioreceptor assay, and receptor epitope reactivity. The sensitivity of immunoblotting was limited by reduced transfer of purified receptor from the gel. However, in addition, the inability to immunoblot the purified receptor with a majority of Graves' sera, under conditions designed to enhance receptor renaturation, appears to reflect a strict conformational requirement for immunoreactivity. Immunoblotting of purified receptors therefore has a limited application in detecting, and defining the epitope reactivity of, TSH receptor autoantibodies.
Leedman
P J
PJ
Burnet Clinical Research Unit, Walter & Eliza Hall Institute of Medical Research, Victoria, Australia.
Harrison
P J
PJ
Harrison
L C
LC
D000818:Animals; D001323:Autoantibodies / Q000097:blood*; D000939:Epitopes; D006111:Graves Disease / Q000276:immunology; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D011989:Receptors, Thyrotropin / Q000276:immunology*; D012680:Sensitivity and Specificity; D013552:Swine; D013961:Thyroid Gland / Q000276:immunology
D001323:Autoantibodies; D000939:Epitopes; D011989:Receptors, Thyrotropin