Dopamine-Based Paper Analytical Device for Truly Equipment-Free and Naked-Eye Biosensing Based on the Target-Initiated Catalyzed Oxidation

The development of low cost, portable, and disposable biosensors for equipment-free and naked-eye biosensing is in eager demand for their widespread application in biomedical field, but it is still a challenge. Herein, we propose a novel paper analytical device (PAD) for truly equipment-free and naked-eye biosensing using dopamine as the chromogenic agent based on target-initiated catalyzed oxidation reaction. The dopamine-functionalized PAD (DPAD) possesses a significant three-dimensional net structure, excellent hydrophilicity, and unique response toward G-quadruplex DNAs against other DNAs, benefiting the bio/chemo reaction occurrence to assay target biomolecules. In light of the exceptional properties, the fabricated DPAD was applied in the analysis of Dam MTase through target-triggered exponential isothermal amplification. The recognition and methylation of H1 by Dam MTase contribute to formation of abundant hemin/G-quadruplexes, which catalyze oxidation of dopamine into dopachrome and reduce the dopamine amount on the DPAD surface. In comparison with the case in which Dam MTase is absent, an evident deep pink signal originating from dopachrome is observed directly by the naked eye and relied on Dam MTase concentrations. Therefore, truly equipment-free and naked-eye detection of Dam MTase is achieved with a detection limit of 1.46 U/mL. The fabricated DPAD not only achieves Dam MTase-visualized detection but also permits the accurate determination of other analytes by varying recognizable DNA's sequences, thus offering a universal biosensor and depicting significant potential for widespread applications in biomedical field.