Citation: Fan Lei, Jiang Qunying, Pan Min, Wang Wenxiao, Zhang Li, Liu Xiaoqing. Dual-Mode Sensing of Biomarkers by Mimic Enzyme-Natural Enzyme Cascade Signal Amplification[J]. Acta Chimica Sinica, ;2020, 78(5): 419-426. doi: 10.6023/A20030079 shu

Dual-Mode Sensing of Biomarkers by Mimic Enzyme-Natural Enzyme Cascade Signal Amplification

College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072

Corresponding author: Liu Xiaoqing, xiaoqingliu@whu.edu.cn
Received Date: 19 March 2020
Available Online: 6 May 2020
Fund Project: Project supported by the National Natural Science Foundation of China (No. 81602610) and the Fundamental Research Funds for the Central Universities (No. 2042018kf1006)the National Natural Science Foundation of China 81602610the Fundamental Research Funds for the Central Universities 2042018kf1006

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Highly sensitive and accurate analysis of significant biomarkers such as alkaline phosphatase (ALP) is essential for early detection and treatment of diseases. In this work, a fluorescence/UV-vis dual-mode sensing platform was constructed for amplified detection of ALP and pyrophosphate ion (PPi) based on mimic enzyme-natural enzyme cascade reactions. Cu-Based metal-organic frameworks HKUST-1 which possesses of oxidase-like activity and can effectively catalyze the oxidation of indicator o-phenylenediamine (OPD) by the surface-active sites were prepared. The oxidation products of OPD exhibit strong UV-vis absorption and fluorescent signals at 416 and 568 nm, respectively. After adding PPi, the catalytic activity of HKUST-1 was selectively inhibited due to the combination of PPi with Cu²⁺ on the surface of HKUST-1, that resulted in fluorescence and UV-vis signal reducing. Once ALP was introduced into the system, PPi can be specifically hydrolyzed into phosphate ions (Pi), and the oxidase-like activity of HKUST-1 recovered. Thus, the fluorescent and UV-vis signals were regenerated by an ALP-triggered mimic enzyme-natural enzyme cascade reaction. On account of the inhibition of oxidase-like activity of HKUST-1 by PPi and the recovery by ALP, an ultrasensitive dual-mode sensing platform of biomarkers based on mimic enzyme-natural enzyme cascade reactions has been developed. Under optimal conditions, the linear range of ALP by fluorescence/UV-vis detection is 0.02~3.5 and 0.04~3.5 nmol·L^-1, and the detection limit of fluorescence and UV-vis assay is as low as 0.0078 and 0.039 nmol·L^-1, respectively. As far as we know, it is the first time that the mimic enzyme-natural enzyme cascade reaction is applied to dual-mode bioanalysis. Due to the enzyme cascade amplification and dual-mode signal output, this developed strategy has the advantages of high sensitivity, low detection limit, high accuracy and reliability, and can realize ultrasensitive analysis of ALP in human serum samples, which shows great potential for clinical diagnosis.
- biomarker detection,
- alkaline phosphatase,
- dual-mode detection,
- mimic enzyme,
- biosensing
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