In Situ Pt Staining Method for Simple, Stable, and Sensitive Pressure-Based Bioassays
2018
Li, Jiuxing | Liu, Fang | Zhu, Zhi | Liu, Dan | Chen, Xiaofeng | Song, Yanling | Zhou, Leiji | Yang, Chaoyong
Pressure-based bioassays (PASS) integrate a molecular recognition process with a catalyzed gas generation reaction, enabling sensitive and portable quantitation of biomarkers in clinical samples. Using platinum nanoparticles (PtNPs) as a catalyst has significantly improved the sensitivity of PASS compared with protein enzyme-based detection. However, PtNPs are easily deactivated during storage or after being decorated with antibodies. Moreover, nonspecific adsorption of PtNPs on substrates has been a problem, resulting in significant backgrounds. To solve these problems of PtNP-based detection, we report a robust, simple, stable, and sensitive Pt staining method for PASS. Detection antibody-decorated gold nanoparticles (AuNPs) are used to perform enzyme-linked immunosorbent assay, followed by Pt staining to stain AuNPs with Ag and Pt bimetallic shells (Au@AgPtNPs), which endow AuNPs with catalytic activity. The concentration of targets can be quantitatively determined by measuring the pressure due to O₂ gas (g) formed by the decomposition of H₂O₂ catalyzed by Au@AgPtNPs. C-reactive protein and avian influenza hemagglutinin 5 neuraminidase 1 can be quantitatively detected with detection limits of 0.015 and 0.065 ng/mL, respectively. The simple, stable, and sensitive properties of the Pt staining-based method will largely broaden the applications of PASS in clinical diagnosis and biomedicine.
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