Magnetic particle-based immunoassay of phosphorylated p53 using protein cage templated lead phosphate and carbon nanospheres for signal amplification

Phosphorylated p53 at serine 15 (phospho-p5315) is a potential biomarker of gamma-radiation exposure. In this paper, we described a new magnetic particle (MP)-based electrochemical immunoassay of human phospho-p5315 using carbon nanospheres (NS) and protein cage nanoparticles (PCN) for signal amplification. Greatly enhanced sensitivity was achieved for three reasons: 1) PCN and the p5315 signal antibody (p5315 Ab2) are linked to the carbon NS (PCN-p5315 Ab2-NS) as a label; 2) PCN increases the amount of metal ions in the cavity of each apoferritin; 3) MPs capture a large amount of primary antibodies. Protein cage templated metallic phosphates, instead of enzymes, as multi-labels have the advantage of eliminating the addition of mediator or immunoreagents and, thus, makes the immunoassay system simpler. Subsequent stripping voltametric analysis, detected olead ions on a disposable screen-printed electrode. The response current was proportional to the phospho-p5315 concentration in the range of 0.02 to 20 ng mL−1 with a detection limit of 0.01 ng mL−1, which was 30-fold lower than that of the ELISA measurement of phospho-p5315. This method shows an acceptable stability and reproducibility and the assay results for phospho-p5315-spiked human serum presented good recovery rates.

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