We developed a triple-readout probe for colorimetric, fluorescent, and fluorescence-lifetime sensing of alkaline phosphatase (ALP) through the hydrolyzed ascorbic acid phosphate (AAP)-mediated formation of silver nanoparticles (AgNPs) on Ag+-deposited MoS2 quantum dots (QDs). Ag+ ions were self-assembled on a monolayer MoS2 QD surface through the formation of Ag-S bonds. When ALP hydrolyzed AAP in an alkaline buffer, the resultant ascorbic acid (AA) triggered the reduction of the bound Ag+ ions into AgNPs on the MoS2 QD surface. The resultant AgNPs induced an efficient fluorescence quenching of the MoS2 QDs through simultaneous static and dynamic quenching processes, generated an intense surface plasmon resonance peak, and triggered a reduction in the fluorescence lifetime of the MoS2 QDs. Electron microscopy and spectroscopic techniques revealed the successful fabrication of Ag+-deposited MoS2 QDs and the ALP-mediated formation of AgNPs on the MoS2 QD surface. The linear quantification ranges for ALP were 0.05-2.5, 0.1-4, and 1-4 units L-1 in the fluorescent, colorimetric, and fluorescence-lifetime detection modes, respectively. In addition, the proposed probe integrated with an ALP-linked sandwich immunoassay exhibited high sensitivity and selectivity for the fluorescence sensing of rabbit immunoglobulin G with a detection limit of 8 pg mL-1 and linear range of 25-1000 pg mL-1. The sensitivity of the probe is comparable to those of previously reported immunoassays involving ultrasensitive electrochemical detection, hydrogen evolution reactions, or electron spin resonance. The probe integrated with the sandwich assay serves as a promising platform for the detection of target proteins in clinical samples.
Keywords: Alkaline phosphatase; Immunoassay; MoS2 quantum dots; Silver nanoparticles; Triple-modal sensors.
© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.