Immune Complex Transfer Enzyme Immunoassay for Anti-Thyroglobulin IgG Using 2,4-Dinitrophenyl-thyroglobulin, Biotinyl-thyroglobulin and Streptavidin-β-D-galactosidase Conjugate
作者:
Yoshikuni Yogi,
Takeyuki Kohno,
Kouichi Hirota,
Hirotaka Toshimori,
Shigeru Matsukura,
Toshiaki Setoguchi,
Eiji Ishikawa,
期刊:
Analytical Letters
(Taylor Available online 1993)
卷期:
Volume 26,
issue 10
页码: 2143-2159
ISSN:0003-2719
年代: 1993
DOI:10.1080/00032719308017458
出版商: Taylor & Francis Group
关键词: Enzyme immunoassay;Anti-thyroglobulin IgG;Biotin;Streptavidin;β-D-Galactosidase
数据来源: Taylor
摘要:
A sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for anti-thyroglobulin IgG in serum is described. Anti-thyroglobulin IgG in serum was reacted simultaneously with 2,4-dinitrophenyl-thyroglobulin and biotinyl-thyroglobulin. The immune complex formed of the three components was trapped onto polystyrene balls coated with (anti-2,4-dinitrophenyl group) IgG and, after washing, reacted with streptavidin-β-D-galactosidase conjugate. After washing, the immune complex was eluted from the polystyrene balls with ϵN-2,4-dinitrophenyl-L-lysine and transferred to polystyrene balls coated with (anti-human IgG γ-chain) IgG. β-D-Galactosidase activity bound to the last polystyrene balls was assayed by fluorometry. Biotinylthyroglobulin and streptavidin-β-D-galactosidase conjugate could be prepared more easily than thyroglobulin-β-D-galactosidase conjugate used in the previous immunoassay. Inactive β-D-galactosidase, used to eliminate interference by anti-β-D-galactosidase antibodies in the previous immunoassay, was not required. The present immunoassay was 300-fold more sensitive than the conventional enzyme immunoassay, although 10-fold less sensitive than the previous immunoassay. Antithyroglobulin IgG was demonstrated in all patients with autoimmune thyroid diseases and 54% of healthy subjects.
点击下载:
PDF (565KB)
返 回