Novel and Sensitive Enzyme Immunoassay for Anti-Thyroglobulin Antibodies in Serum Using Dinitrophenyl Thyroglobulin and Thyroglobulin-Peroxidase Conjugate

Abstract
A novel and sensitive enzyme immunoassay for anti-thyroglobulin antibodies is described. Anti-thyroglobulin antibodies in test serum were reacted simultaneously with dinitrophenyl thyroglobulin and thyroglobulin-horseradish peroxidase conjugate. The complex formed of anti-thyroglobulin antibodies, dinitrophenyl thyroglobulin and thyroglobulin-peroxidase conjugate was trapped onto two affinity-purified rabbit (anti-dinitrophenyl bovine serum albumin) IgG-coated polystyrene balls. Peroxidase activity bound to the polystyrene balls was assayed by fluorimetry using 3-(4-hydroxyphenyl)propionic acid as substrate. This assay was more sensitive than the conventional enzyme immunoassays, and anti-thyroglobulin antibodies were demonstrated in part of healthy subjects and in all patients with Graves' disease and chronic thyroiditis.

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