蒋金和
云师大化工学院杨通博士在top期刊《Biosensors and Bioelectronics》发表最新研究成果
2023-12-10 21:27
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2023年12月8日,Elsevier 旗下top期刊《Biosensors and Bioelectronics》在线发表了云南师范大学化学化工学院杨通博士团队的最新研究成果《Nanoplasmonic biosensors for multicolor visual analysis of acetylcholinesterase activity and drug inhibitor screening in point-of-care testing》。云南师范大学化学化工学院为唯一作者兼通讯作者单位

https://www.sciencedirect.com/science/article/abs/pii/S0956566323008540


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Abstract

The monitoring of acetylcholinesterase (AChE) activity and the screening of its inhibitors are significance of the diagnosis and drug therapy of nervous diseases. A metal ions-mediated signal amplification strategy was developed for the highly sensitive and multicolor assay of AChE activity and visually screening its drug inhibitors. After the specific reaction between AChE and acetylthiocholine (ATCh), the hydrolysis product thiocholine (TCh) can directly and decompose the α-FeOOH nanorods (NRs) to release amounts of Fe2+, which was regarded as Fenton reagent to efficiently catalyze H2O2 to produce ·OH. Then, the as-formed ·OH can further largely shorten the gold nanobipyramids (Au NBPs), generating a series of palpable color variations. The linear range for AChE activity was 0.01–500.0 U/L with the limit of detection as low as 0.0074 U/L. The vivid visual effects could be easily distinguished for the multicolor assay of AChE activity by naked eye in visible light. To achieve the point-of-care testing, Au NBPs were further assembled on polymeric electrospun nanofibrous films (ENFs) surface as test strips for the easy-to-use test of AChE activity by RGB values with a smartphone. Fascinatingly, this proposed strategy can be used for the visual screening AChE inhibitors or non-inhibitors. Comparing with the clinical drugs (rivastigmine tartrate, and donepezil), some natural alkaloids such as evodiamine, caffeine, camptothecin, and berberine hydrochloride were selected as inhibitor modes to confirm the drug screening capability of this method. This proposed strategy may have great potential in the other disease-related enzymatic biomarkers assay and the rapid screening of drug therapy.

Introduction

Signal amplification have been regarded as an effective strategy to improve the sensitivity of the very low content levels of biomolecules, such as biological enzyme, protein, incretion, nucleic acids, and metabolites(Wu et al. 2019, 2020). To date, many typical types of amplification strategies including enzymatic reaction,(Liu et al., 2018) chemical redox-cycling,(Chen et al., 2019) nanozyme-mediated catalytic reaction(Li et al., 2022b), DNA nanotechnology-mediated cycling (DNA nanomachines, hairpin DNA cascade amplification, DNAzyme-assistant, DNA origami, and so forth),(Dong et al., 2020; Gong et al., 2015; Li et al., 2021) and metal ions-mediated amplification,(Xianyu et al., 2014) have been successfully used to enhance the sensitivity of the above biomolecules. Wherein, metal ions-mediated signal amplification strategy gradually arouses the extensive research interests because of the wide sources of metal ions, the direct interactions and rapid response between metal ions and signal probes. For example, Cu2+ released from CuO nanoparticles directly induced the “click chemistry” reaction,(Qu et al., 2011) the interdiction of fluorescence resonance energy transfer (FRET) process,(Yang et al., 2018) or the activation of the signal-off photoelectrochemical immunoassay,(Luo et al., 2019) for achieving the sensitive detection of antibodies or proteins. With the nanoplasmonic feature of gold nanostructures, the Fe2+/H2O2 or Ag+/reductive products were introduced in analytical systems, which can selectively corrode the gold nanorods (Au NRs),(Liu et al., 2020; Ye et al., 2022) or deposited on gold nanobipyramids (Au NBPs),(Chen et al., 2018; Zhang et al., 2016) to develop the multicolor-based biosensing platforms. In addition, many achievements including the cation-exchange,(Zhang et al., 2016) the interconversion of oxidation-reduction metal ion pairs,(Chen et al., 2018) and metal ions-implanted loop DNA structures,(Chen et al., 2021) have been made to establish some sensitive and versatile biosensors. Comparing with the extra introduction of metal ions, the metal ions from nanomaterials themselves can more directly contact with signal probes, avoiding the extra introduction of redox reagents to simplify some tedious steps. Therefore, it is a more expected that the final products from target analytes and substrates could directly react with metal-based nanomaterials to release numerous metal ions, which would further rapidly and largely induce the signal output variations.

Acetylcholinesterase (AChE), as a vital biological enzyme in cholinergic nervous system, can hydrolyze neurotransmitter acetylcholine (ACh) into acetic acid and choline, which dynamically modulate the level of ACh(Wang et al., 2022). Either too higher or too lower AChE activity in vivo would cause some neurological disease. It is generally recognized that the loss of ACh with the amount of AChE will cause some neurodegenerative disease such as Parkinson, Alzheimer, or Huntington(Liu et al., 2021). On the contrary, the accumulation of ACh in body will result in the neurotransmission disorder and eventual death(Zhang et al., 2023). Therefore, it is very important for the monitoring of AChE activity and the screening of its inhibitors in the diagnosis and drug therapy of nervous diseases. Currently, varieties of sensing strategies have been developed for the testing AChE activity, or further screening of its inhibitor, including colorimetric assay,(Wang et al., 2022; Zhang et al., 2023) fluorescent assay,(Zhang et al., 2022; Zhao et al., 2023) chromatography-mass spectrometry combined technique (HPLC-HRMS),(Stuetz et al., 2020) electrochemical sensor,(Li et al., 2020) and so on. Among them, visually colorimetric analysis has been the focus of considerable popular in the assay of AChE activity because of their convenient, low cost, rapid readout, naked-eyes recognition, and great potential in point-of-care or ready-to-use testing(Liu et al., 2022; Wei et al., 2021). However, much effort has focused on the monochromatic analysis by utilizing the single wavelength characteristic of colorants or fluorophores (dyes, pigments, or oxidates from substrate-enzyme interactions), whose poor visual differentiation, low extinction coefficients, and unsatisfactory sensitivity still limited the dynamic monitoring of AChE activity and its visual screening application toward inhibitors.

By comparison, dual- or multi-colors-based colorimetric or fluorescent analysis could provide more vivid visual discrimination. On the one hand, the cutoff values or ranges of target analytes could be recognized with human vision response, which is beneficial for the rapidly semi-quantification of targets(Li et al., 2022a; Wang et al., 2017). On the other hand, the signal output of multicolor could be easily recorded from a photograph taken by a smartphone, whose colorimetric APP could directly measure their RGB values(Wang et al. 2019a, 2019b; Wei et al., 2021). Therefore, it is more valuable for monitoring AChE activity and visually screening inhibitors or non-inhibitors to construct a multicolor and versatile colorimetric POCT technique. Furthermore, with the development of pharmaceutical chemistry, visual drug screening is of great significance to rapidly screen new effective ingredients from chemical synthetic drugs or natural products in the course of new drug discovery.

In 2021, Song et al. reported a fluorescent sensing platform for the AChE assay with silicon quantum dots (QDs) as a fluorescence (FL) oscillator and FeOOH nanostructures as a quencher(Song et al., 2021). AChE reacted with acetylthiocholine (ATCh) to produce the enzymatic hydrolysate TCh, which can directly and decompose the FeOOH to release Fe2+ to restore the FL of Si QDs. Inspired by this work and the above recent research progress, an α-FeOOH nanorods (NRs)-mediated multicolor nanoplasmonic biosensor toward AChE activity evaluation and its inhibitors screening by an easy-to-use smartphone was established. As seen Scheme 1, making full use of the interaction between TCh and α-FeOOH NRs to release amount of Fe2+, Fe2+ as Fenton reagent can efficiently catalyze H2O2 to produce ·OH. Then, the ·OH could corrode the Au NBPs to induce the blue shifts of their longitudinal localized surface plasmon resonance (LSPR) absorption wavelength, resulting in the abundant color variations from brown, green, cyan, mazarine, violet, and pink in homogenous solution with the AChE varying activities. Furthermore, functional electrospun nanofibrous films (ENFs) have been regarded as a versatile test strip for establishing a point-of-care testing (POCT) platform, which also exhibited some easy-to-use sensing performances(Yang et al., 2020). Therefore, the Au NBPs were assembled on polymeric ENFs surface to develop a POCT test strip for the solid phase detection of AChE activity and its inhibitor screening with a smartphone platform。

扩展阅读:

杨通                              

男,19905月生,汉族,中共党员,博士研究生,分析化学专业,硕士研究生导师,云南省“兴滇英才支持计划”——青年人才

邮箱yangtong2018@ynnu.edu.cnyt09132149@163.com


主要研究方向

研究方向:化学与生物传感、纳米分析化学、功能性核酸传感技术、光谱分析及电分析化学

研究专长:功能性电纺纳米结构、多孔框架纳米结构的制备及其在可视化、光电或光热生化传感中的应用

研究业绩:近些年来,主持国家自然科学基金项目1项、云南省科技厅基础研究计划项目2云南省教育厅科研项目1项。以第一作者和通讯作者身份,已在Anal. Chem.Biosens. Bioelectron.Sens. Actuators, BFood Chem.TrAC, Trends Anal. Chem.ACS Appl. Mater. InterfacesNanoscale等分析化学、材料化学相关领域期刊,共发表SCI论文、国内核心期刊论文20余篇。授权国家发明专利1项。


主要招生专业

分析化学专业


教育与工作经历

201809月~至今云南师范大学 化学化工学院 应用化学系

201906月~202105月,云南省德宏州梁河县平山乡 驻村参与国家脱贫攻坚任务

201309月~201806月,西南大学 药学院 分析化学专业 博士/研究生(硕博连读)

200909月~201306月,曲靖师范学院 化学化工学院 应用化学专业 学士/本科


获奖成果和荣誉称号

1. 杨通 云南省兴滇英才支持计划”——青年人才,2019

2. 2022年指导马璐瑶等本科生参加微瑞杯第三届全国大学生化学实验创新设计大赛荣获西南赛区一等奖、全国总决赛一等奖(国家级A类赛事)

3. 2022年指导周爱齐等本科生参加第八届云南省互联网+”大学生创新创业大赛荣获铜奖


主持科研项目

1. 国家自然科学基金项目青年基金项目,基于电纺纤维固相传感平台的肿瘤标志物可视化分析方法与技术,项目批准号:21904114,起止年限:2020/01—2022/12

2. 云南省基础研究计划项目面上项目,基于DNA纳米机器的功能性电纺纤维生物传感膜的分析方法与技术,项目批准号:202201AT070028,起止年限:2022/06—2025/05

3. 云南省基础研究计划项目青年项目,静电纺纳米纤维传感界面的功能性调控及其在生物检测中的应用,项目批准号:202001AU070067,起止年限:2020/06—2023/05

4. 云南省教育厅科学研究项目,荧光电纺纤维固相膜传感平台在生化分析中的应用,项目批准号:2019J0066,起止年限:2019/06—2020/05


代表性著作、论文、专利等

论文

1. Tong Yang; Chun Mei Li; Jia Hui He; Bin Chen; Yuan Fang Li; Cheng Zhi Huang*. Ratiometrically Fluorescent Electrospun Nanofibrous Film as a Cu2+-Mediated Solid-Phase Immunoassay Platform for Biomarkers. Anal. Chem. 201890, 9966-9974. (中国科学院:一区IF2022=8.008)

2. Mei Li1; De Yan Li1; Zi Ying Li; Rong Hu; Yun Hui Yang*; Tong Yang*. A Visual Peroxidase Mimicking Aptasensor Based on Pt Nanoparticles-Loaded on Iron Metal Organic Gel for Fumonisin B1 Analysis in Corn Meal. Biosens. Bioelectron. 2022, 209, 114241. (中国科学院:一区IF2022=12.545)

3. De Yan Li; Zi Ying Li; Lei Han; Shuang Meng; Rong Hu; Yun Hui Yang; Tong Yang*. Target-Induced Tripedal G-Quadruplex DNAzyme for Multicolor Visual Point-of-Care Testing of Biomarkers Using Au Nanorods-Decorated Electrospun Nanofibrous Films. Sens. Actuators, B 2022, 371, 132510. (中国科学院:一区IF2022=9.221)

4. Tong Yang*; Lei Zhan.; Cheng Zhi Huang**. Recent Insights into Functionalized Electrospun Nanofibrous Films for Chemo-/Bio-Sensors. TrAC, Trends Anal. Chem. 2020124, 115813. (中国科学院:一区IF2022=14.908)

5. Yan Guan; Peng Bin Si; Tong Yang*; Yuan Wu*; Yun Hui YangRong Hu*. A Novel Method for Detection of Ochratoxin A in Foods—Co-MOFs Based Dual Signal Ratiometric Electrochemical Aptamer Sensor Coupled with DNA Walker. Food Chem. 2023, 403, 134316. (中国科学院:一区IF2022=9.231)

6. Zi Ying LiDe Yan Li; Long Huang; Rong Hu; Tong Yang*Yun Hui Yang**. An Electrochemical Aptasensor Based on Intelligent Walking DNA Nanomachine with Cascade Signal Amplification Powered by Nuclease for Mucin 1 Assay. Anal. Chim. Acta 2022, 1214, 339964. (中国科学院:一区IF2022=6.911)

7. Yan Guan; Fu Peng Wang; Zhi Xiong Chen; Yun Hui Yang; Tong Yang*; Rong Hu**. Ratiometrically Homogeneous Electrochemical Biosensor Based on the Signal Amplified Strategy of Dual DNA Nanomachines for MicroRNA Analysis. Talanta 2023, 254, 124191. (中国科学院:一区IF2022=6.556)

8. Jia Li Liu1; Yu Chan Ma1Tong Yang*; Rong Hu**; Yun Hui Yang***. A Single Nucleotide Polymorphism Electrochemical Sensor Based on DNA-Functionalized Cd-MOFs-74 as Cascade Signal Amplification Probes. Microchim. Acta 2021, 188(8), 266. (中国科学院IF2022=6.408)

9. Tong Yang; Peng Hou; Lin Ling Zheng.; Lei Zhan; Peng Fei Gao; Yuan Fang Li;* Cheng Zhi Huang*. Surface-Engineered Quantum Dots/Electrospun Nanofibers as A Networked Fluorescent Aptasensing Platform toward Biomarkers. Nanoscale 2017917020-17028. (中国科学院IF2022=8.307)

10. Tong Yang; Jun Ma; Shu Jun Zhen;* Cheng Zhi Huang*. Electrostatic Assembly of Well-Dispersed AgNPs on the Surface of Electrospun Nanofibers as Highly Active SERS Substrates for Wide-Range pH Sensing. ACS Appl. Mater. Interfaces 20168, 14802-14811. (中国科学院IF2022=10.383)

11. Tong Yang; Hong Yan Zou; Cheng Zhi Huang*. Synergetic Catalytic Effect of Cu2–xSe Nanoparticles and Reduced Graphene Oxide Coembedded in Electrospun Nanofibers for the Reduction of a Typical Refractory Organic Compound. ACS Appl. Mater. Interfaces 20157, 15447-15457. (中国科学院IF2022=10.383)

12. Tong Yang; Hui Yang; Shu Jun Zhen;* Cheng Zhi Huang*. Hydrogen-Bond-Mediated in Situ Fabrication of AgNPs/Agar/PAN Electrospun Nanofibers as Reproducible SERS Substrates. ACS Appl. Mater. Interfaces 20157, 1586-1594. (中国科学院IF2022=10.383)

13. 李紫滢李德燕杨建梅胡蓉杨通*; 杨云慧基于FRET-DNA纳米机器的循环信号放大策略用于检测前列腺特异性抗原分析化学 2022, 50 (7), 1032-1040.

14. 牛鹏博李德蕾胡蓉杨通*杨云慧*. 基于Cu2+诱导信号放大策略的电化学适体传感器用于检测赭曲霉毒素A.分析试验室 2023, 42 (5), 595-600.

15.马璐瑶; 张钧富; 陈怡橦; 杨*. RGB阵列式荧光传感技术用于检测高原湖泊中多种重金属离子. 大学化学 2023, 38(4), 78-87. (教改论文)

专利

1. 杨通、梁宇、黄承志,专利名称:一种具有铜离子响应性的电纺纤维膜的制备方法及产品和利用其检测铜离子的方法,中国发明专利,专利授权号:ZL 202011198140.6,授权日期:2022/07


主讲课程

本科生课程《分析化学《仪器分析》《分析化学实验》《仪器分析实验》《药物化学》

研究生课程《光谱分析与电化学分析《现代化学化工前沿进展》



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