Development of MnO2 nanosheets nanozyme-based colorimetric and photothermal dual-signal platform for monitoring alkaline phosphatase activity

Herein, a dual-signal biosensing platform for monitoring alkaline phosphatase (ALP) activity was developed. This platform combined the oxidase-mimicking activity of MnO2 nanosheets (NS) with the colorimetric and photo- thermal properties of oxidized 3,3 ',5,5 '-tetramethylbenzidine (oxTMB). MnO2 NS effectively catalyzed the oxidation of TMB to oxTMB, resulting in a color change from colorless to blue and a significant light absorption band at 652 nm. Meanwhile, oxTMB demonstrated pronounced photothermal properties, with the solution exhibiting a notable increase in temperature upon irradiation with an 808 nm laser. In the presence of ALP, 2Phospho-L-ascorbic acid trisodium salt (AAP) was converted into L-ascorbic acid (L-AA), which then reacted with MnO2 NS. Consequently, MnO2 NS was inhibited from oxidizing TMB to oxTMB. Based on this principle, a dual- readout assay that integrated both colorimetric and photothermal signals for ALP detection was developed. The detection limits were 0.08 mU/mL for the colorimetric method and 0.12 mU/mL for the photothermal method. Furthermore, the platform was successfully applied to human serum samples, demonstrating its effectiveness in detecting ALP in complex biological matrices. The combination of colorimetric and photothermal responses enhanced both the accuracy and reliability of ALP detection, making this platform a promising tool for biomedical applications.