The application of the QDs/H2O2 chemiluminescence system in HRP assay and HRP-based immunoassay

Abstract

Horseradish peroxidase (HRP) is usually used as a label enzyme in immunoassay so the method used for HRP detection in enzyme immunoassay (EIA) plays a key role in sensitivity and precision. The catalytic activity of HRP does not strictly follow classic Michaelis-Menten kinetics, probably due to the inactivation of the enzyme at high concentrations of H2O2. In this paper, a highly sensitive alternative procedure for the HRP assay using H2O2-sensitive CdTe quantum dots as a chemiluminescence (CL) system is reported. This method can measure a much more accurate and reliable value of Km (187 mM H2O2) in comparison with the standard detection method. This system also was applied to thyroid hormone (T4) detection using HRP-based immunoassay. The QD/H2O2 system exhibits a higher linear range of 0.2–16 μg/dL with the improved LOD value of 0.06 μg/dL and selective response to T4, which was better than the commercial colorimetric immunoassay. Meanwhile, the proposed method has been successfully applied to the clinical determination of T4 in the serum samples, and the results confirmed an excellent correlation with the conventional ELISA method (R2 = 0.9832), indicating the potential applications of the method for clinical diagnosis as well.