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

作　　者：Yujie Sun Lei Wen Huili Ma Wenlin Ma Zhenqian Fu Yinhui Li Chengwu Zhang Lin Li Jinhua Liu 

机构地区：[1]Institute of Advanced Materials(IAM),Key Laboratory of Flexible Electronics(KLOFE),Nanjing Tech University,Nanjing 211816,China [2]Key Laboratory for Green Organic Synthesis and Application of Hunan Province,Key Laboratory of Environmentally Friendly Chemistry,Application of Ministry of Education,College of Chemistry,Xiangtan University,Xiangtan 411105,China [3]School of Basic Medical Sciences,Shanxi Medical University,Taiyuan 310003,China

出　　处：《Chinese Chemical Letters》2023年第4期444-448,共5页中国化学快报（英文版）

基　　金：the National Natural Science Foundation of China (Nos. 22174065, 21974119);the Science and Technology Planning Project of Hunan Province (No. 2020RC_(3)046);Hunan Provincial Natural Science Foundation of China (No.2019JJ30020)。

摘　　要：Mulit-enzyme cascades are a major type of chemical transformations and play a crucial role in biological signal transduction and metabolism. Herein, a trienzyme cascade-triggered fluorescent immunosensor 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 highresolution mass spectrum,1H NMR,^(13)C NMR and theoretical calculations. The integration of the twoenzyme 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(c Tn I) as the model antigen, a trienzyme cascade-triggered fluorescent immunosensor platform was developed for quantitative detecting c Tn I in a wide linear range from 2 ng/m L to 150 ng/m L with a detection limit of 0.67ng/m L. In addition, the proposed platform was successfully applied in detection of c Tn I in serum of clinical patients. Overall, the developed fluorescent immunosensor performs powerful implications for researching enzyme cascade systems in the field of biomedicine.

关 键 词：Enzyme cascade Alkaline phosphatase TYROSINASE Horseradish peroxidase Cardiac troponin I 

分 类 号：O657.3[理学—分析化学] R313[理学—化学]