Electrochemical Immunoassay of E. coli in Urban Sludge Using Electron Mediator-mediated Enzymatic Catalysis and Gold Nanoparticles for Signal Amplification

doi:10.1007/s40242-017-7254-z

高等学校化学研究 ›› 2018, Vol. 34 ›› Issue (1): 101-106.doi: 10.1007/s40242-017-7254-z

Electrochemical Immunoassay of E. coli in Urban Sludge Using Electron Mediator-mediated Enzymatic Catalysis and Gold Nanoparticles for Signal Amplification

LU Wenjie¹, XU Rongjin¹, ZHANG Xinai², SHEN Jianzhong¹, LI Changfeng¹

1. School of Energy and Power Engineering, Jiangsu University, Zhenjiang 212013, P. R. China;
2. School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, P. R. China

收稿日期:2017-07-31 出版日期:2018-02-01 发布日期:2018-01-20
通讯作者: ZHANG Xinai,E-mail:zhangxinai@mail.ujs.edu.cn;LI Changfeng,E-mail:cfli@ujs.edu.cn E-mail:zhangxinai@mail.ujs.edu.cn;cfli@ujs.edu.cn
基金资助:
Supported by the National Natural Science Foundation of China(Nos.21205051, 51608235) and the Key Project of the Ministry of Education of China(No.210078).

Electrochemical Immunoassay of E. coli in Urban Sludge Using Electron Mediator-mediated Enzymatic Catalysis and Gold Nanoparticles for Signal Amplification

LU Wenjie¹, XU Rongjin¹, ZHANG Xinai², SHEN Jianzhong¹, LI Changfeng¹

1. School of Energy and Power Engineering, Jiangsu University, Zhenjiang 212013, P. R. China;
2. School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, P. R. China

Received:2017-07-31 Online:2018-02-01 Published:2018-01-20
Contact: ZHANG Xinai,E-mail:zhangxinai@mail.ujs.edu.cn;LI Changfeng,E-mail:cfli@ujs.edu.cn E-mail:zhangxinai@mail.ujs.edu.cn;cfli@ujs.edu.cn
Supported by:
Supported by the National Natural Science Foundation of China(Nos.21205051, 51608235) and the Key Project of the Ministry of Education of China(No.210078).

摘要/Abstract

摘要： An electrochemical immunosensor was developed for sensitive assay of E. coli in urban sludge, in which electron mediator-mediated enzymatic catalysis and gold nanoparticles(AuNPs) were utilized for signal amplification. The immnuosensing platform chitosan-thionine(chit-thio)/poly(amidoamine) dendrimer-encapsulated AuNPs [PAMAM(Au)] composites were first prepared from chit-thio and PAMAM(Au) using the layer-by-layer method to provide a matrix for high-stability and high-bioactivity bindings of the capture antibody(_cAb). Moreover, the {_dAb-AuNPs-HRP} nanoprobes were designed to exploit the amplification effect of the carrier AuNPs due to the loading amounts of horseradish peroxidase(HRP) and the detection antibody(_dAb). The sandwich-type immunoassay was then successfully used to assay E. coli based on the oxidation of thionine as a result of H₂O₂-induced enzymatic catalytic reaction by HRP. This study presents a powerful tool in electrochemical immunoassay for E. coli detection with rapid response, high-sensitivity and high-specificity, providing a potential new tool for feasibility assessment of sludge recycle.

关键词: Electron mediator, Enzymatic catalysis, Electrochemical immunosensor, E. coli, Urban sludge

Abstract: An electrochemical immunosensor was developed for sensitive assay of E. coli in urban sludge, in which electron mediator-mediated enzymatic catalysis and gold nanoparticles(AuNPs) were utilized for signal amplification. The immnuosensing platform chitosan-thionine(chit-thio)/poly(amidoamine) dendrimer-encapsulated AuNPs [PAMAM(Au)] composites were first prepared from chit-thio and PAMAM(Au) using the layer-by-layer method to provide a matrix for high-stability and high-bioactivity bindings of the capture antibody(_cAb). Moreover, the {_dAb-AuNPs-HRP} nanoprobes were designed to exploit the amplification effect of the carrier AuNPs due to the loading amounts of horseradish peroxidase(HRP) and the detection antibody(_dAb). The sandwich-type immunoassay was then successfully used to assay E. coli based on the oxidation of thionine as a result of H₂O₂-induced enzymatic catalytic reaction by HRP. This study presents a powerful tool in electrochemical immunoassay for E. coli detection with rapid response, high-sensitivity and high-specificity, providing a potential new tool for feasibility assessment of sludge recycle.

Key words: Electron mediator, Enzymatic catalysis, Electrochemical immunosensor, E. coli, Urban sludge

LU Wenjie, XU Rongjin, ZHANG Xinai, SHEN Jianzhong, LI Changfeng. Electrochemical Immunoassay of E. coli in Urban Sludge Using Electron Mediator-mediated Enzymatic Catalysis and Gold Nanoparticles for Signal Amplification[J]. 高等学校化学研究, 2018, 34(1): 101-106.

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Electrochemical Immunoassay of E. coli in Urban Sludge Using Electron Mediator-mediated Enzymatic Catalysis and Gold Nanoparticles for Signal Amplification

Electrochemical Immunoassay of E. coli in Urban Sludge Using Electron Mediator-mediated Enzymatic Catalysis and Gold Nanoparticles for Signal Amplification

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