More sensitive and simpler immune complex transfer enzyme immunoassay for antithyroglobulin igg in serum

✍ Scribed by Takeyuki Kohno; Tomohiro Mitsukawa; Shigeru Matsukura; Yoshizo Tsunetoshi; Eiji Ishikawa

• Publisher: John Wiley and Sons
• Year: 1989
• Tongue: English
• Weight: 568 KB
• Volume: 3
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/jcla.1860030306

✦ Synopsis

A more sensitive and simpler immune complex transfer enzyme immunoassay for antithyroglobulin IgG in serum and its use for the measurement of antithyroglobulin IgG in healthy subjects and patients with thyroid diseases are described. Antithyroglobulin IgG in test serum was reacted simultaneously with dinitrophenyl thyroglobulin and thyroglobulinp-D-galactosidase conjugate. The complex formed of antithyroglobulin IgG, dinitrophenyl thyroglobulin, and thyroglobulin-P-D-galactosidase conjugate was trapped onto two polystyrene balls coated with aff inity-purified rabbit (antidinitrophenyl bovine serum albumin) IgG. The polystyrene balls were washed to eliminate nonspecific IgG in the test serum, and the complex was eluted from the polystyrene balls with dinitrophenyl-L-lysine and transferred to two polystyrene balls coated with affinity-purified rabbit (anti-human IgG y-chain) IgG. P-D-Galactosidase activity bound to the polystyrene balls was assayed by fluorimetry using 4-methylumbelliferyl-P-D-galactoside as substrate. As compared with the previous immune complex transfer enzyme immunoassay, the procedure was simplified by reducing one incubation step and one washing step, and the detection limit of antithyroglobulin IgG in serum (0.1 p,g/liter) was lowered 100-fold. More careful and extensive examination than in a previous study revealed the presence of antithyroglobulin IgG in a large proportion of healthy subjects and in all patients with Graves' disease and chronic thyroiditis.