4.7 Article

Engineering trienzyme cascade-triggered fluorescent immunosensor platform by sequentially integrating alkaline phosphatase, tyrosinase and horseradish peroxidase

Journal

CHINESE CHEMICAL LETTERS
Volume 34, Issue 4, Pages -

Publisher

ELSEVIER SCIENCE INC
DOI: 10.1016/j.cclet.2022.06.077 1001-8417

Keywords

Enzyme cascade; Alkaline phosphatase; Tyrosinase; Horseradish peroxidase; Cardiac troponin I

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In this study, a trienzyme cascade-triggered fluorescent immunosensor platform was constructed by integrating alkaline phosphatase (ALP), tyrosinase (TYR), and horseradish peroxidase (HRP). The platform utilized a rapid in situ fluorogenic reaction to produce a yellow fluorescent compound. The developed immunosensor showed significant enhancements in fluorescence and successfully detected a cardiac biomarker in clinical samples.
Mulit-enzyme cascades are a major type of chemical transformations and play a crucial role in biolog-ical signal transduction and metabolism. Herein, a trienzyme cascade-triggered fluorescent immunosen-sor platform was constructed by sequentially integrating alkaline phosphatase (ALP), tyrosinase (TYR) and horseradish peroxidase (HRP). The proposed platform was based on HRP-induced a rapid in situ fluorogenic reaction between dopamine (DA) and 1,5-dihydroxynaphthalene (DHA) to produce a strong yellow azamonardine fluorescent compound (AFC). The obtained AFC was clearly characterized by high -resolution mass spectrum, 1 H NMR, 13 C NMR and theoretical calculations. The integration of the two -enzyme system (TYR and HRP) or three-enzyme system (ALP, TYR and HRP) led to a maximum of 400.0 -fold and 250.0-fold fluorescence enhancements, respectively. Using cardiac troponin I (cTnI) as the model antigen, a trienzyme cascade-triggered fluorescent immunosensor platform was developed for quantita-tive detecting cTnI in a wide linear range from 2 ng/mL to 150 ng/mL with a detection limit of 0.67 ng/mL. In addition, the proposed platform was successfully applied in detection of cTnI in serum of clin-ical patients. Overall, the developed fluorescent immunosensor performs powerful implications for re-searching enzyme cascade systems in the field of biomedicine.(c) 2023 Published by Elsevier B.V. on behalf of Chinese Chemical Society and Institute of Materia Medica, Chinese Academy of Medical Sciences.

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