Abstract
This work reports a novel electrochemical immunoassay protocol with signal amplification for determination of low-abundance protein (free prostate-specific antigen, PSA, used as a model) with high sensitivity and high selectivity by coupling metal sulfide (PbS)-based nanolabels with cleavage of the corresponding lead ion-induced DNAzymes. The assay mainly consists of an antigen-antibody immunoreaction with metal nanolabel in a transparent 96-well polystyrene microplate, the release of metal ions from the nanolabel, and cleavage of metal ion-induced DNAzyme. The signal is amplified by the labeled redox tag (ferrocene) on the DNAzyme-based sensor. In the presence of target analyte, the sandwiched immunocomplex can be formed between the primary antibody on the microplate and the corresponding metal sulfide nanolabel. The carried nanolabel can release numerous metal ions by acid, and induce the cleavage of the corresponding DNAzyme, thus resulting in the change of electrochemical signal. Under optimal conditions, the DNAzyme-based immunoassay presents an obvious electrochemical response for the detection of PSA, and allows detection of PSA at a concentration as low as 0.1 pg mL(-1). Intra-assay and interassay coefficients of variation (CV) were less than 9.5% and 10%, respectively. No significant differences at the 0.05 significance level were encountered in the analysis of 12 clinical serum specimens between the developed immunoassay and a commercially available enzyme-linked immunosorbent assay (ELISA).
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