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A Homogeneous Time-Resolved Fluorescence Immunoassay Method for the Measurement of Compound W
Biao Huang1, Huixin Yu1, Jiandong Bao1
1Research Department, Jiangsu Nuclear Medicine Research Laboratory and Jiangyuan Hospital, Wuxi, China.
Objective:
Using compound W (a 3,3'-diiodothyronine sulfate [T2S] immuno-crossreactive material)-specific polyclonal antibodies and homogeneous time-resolved fluorescence immunoassay assay techniques (AlphaLISA) to establish an indirect competitive compound W (ICW) quantitative detection method.
Method:
Photosensitive particles (donor beads) coated with compound W or T2S and rabbit anti-W antibody were incubated with biotinylated goat anti-rabbit antibody. This constitutes a detection system with streptavidin-coated acceptor particle. We have optimized the test conditions and evaluated the detection performance.
Results:
The sensitivity of the method was 5 pg/mL, and the detection range was 5 to 10 000 pg/mL. The intra-assay coefficient of variation averages <10% with stable reproducibility.
Conclusions:
The ICW-AlphaLISA shows good stability and high sensitivity and can measure a wide range of compound W levels in extracts of maternal serum samples. This may have clinical application to screen congenital hypothyroidism in utero.
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