Abstract
Thyroid-stimulating hormone (TSH) is widely used as a marker of thyroid function. A rapid TSH assay enables diagnosis during the first visit at the doctor’s office aiding to faster and cost-effective medical treatment. To accomplish such an assay method europium(III) chelate nanoparticles were coated with anti-TSH monoclonal antibody. Captured anti-TSH monoclonal antibody was immobilized onto single wells by streptavidin–biotin chemistry and the assay was carried out in dry chemistry format using 5 μl of sample in a 30 μl assay volume in the commercial AiO immunoassay system. The developed TSH nanoparticle assay was performed in a kinetic mode using a 10-min incubation time. The analytical sensitivity of the developed assay was 0.0012 mIU l −1 corresponding to the fourth generation TSH assay and less than 0.02 mIU l −1 when serum-based matrix was used for calibrators. The dynamic range of the assay was more than three orders of magnitude and no high-dose hook effect was observed at 100 mIU l −1 of TSH. Correlation with an automated commercial assay was good ( y = 0.96 ± 0.02, intercept = 0.12 ± 0.09, S y | x = 0.49, R = 0.988). Intra- and inter-assay variations were 4–14 and 6–17%, respectively. The developed quantitative one-step all-in-one dry reagent time-resolved fluorometric immunoassay has great potential for rapid analysis of serum thyroid-stimulating hormone in a point-of-care environment when antibody-coated high specific activity Eu(III) nanoparticles were used as labels.
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