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Electrochemiluminescence Biosensor for Ascorbic Acid Based on Target Transformation of Cell-Free RNA Transcription System and Duplex-Specific Nuclease-Assisted Recycling Amplification

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Download Hi-Res ImageDownload to MS-PowerPointCite This:Anal. Chem. 2024, 96, 44, 17807-17813
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    Electrochemiluminescence Biosensor for Ascorbic Acid Based on Target Transformation of Cell-Free RNA Transcription System and Duplex-Specific Nuclease-Assisted Recycling Amplification
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    • Ming Chen
      Ming Chen
      Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China
      More by Ming Chen
    • Hongfu Zeng
      Hongfu Zeng
      Department of Thoracic Oncology, Clinical Oncology School of Fujian Medical University, Fujian Cancer Hospital (Fujian Branch of Fudan University Shanghai Cancer Center), No. 420 Fuma Road, Fuzhou, Fujian 350014, China
      More by Hongfu Zeng
    • Fang Luo
      Fang Luo
      Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China
      More by Fang Luo
      Orcidhttps://orcid.org/0000-0003-1480-0677
    • Yunjian Huang*
      Yunjian Huang
      Department of Thoracic Oncology, Clinical Oncology School of Fujian Medical University, Fujian Cancer Hospital (Fujian Branch of Fudan University Shanghai Cancer Center), No. 420 Fuma Road, Fuzhou, Fujian 350014, China
      *Email: [email protected]
      More by Yunjian Huang
    • Cuiying Lin*
      Cuiying Lin
      Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China
      *Email: [email protected]
      More by Cuiying Lin
    • Jian Wang
      Jian Wang
      Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China
      More by Jian Wang
      Orcidhttps://orcid.org/0000-0002-3917-9461
    • Bin Qiu
      Bin Qiu
      Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China
      More by Bin Qiu
      Orcidhttps://orcid.org/0000-0003-2011-3320
    • Zhenyu Lin*
      Zhenyu Lin
      Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China
      *Email: [email protected]. Tel.: 86-591-22866135.
      More by Zhenyu Lin
      Orcidhttps://orcid.org/0000-0001-7890-6812
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    Analytical Chemistry

    Cite this: Anal. Chem. 2024, 96, 44, 17807–17813
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    https://doi.org/10.1021/acs.analchem.4c04206
    Published October 27, 2024
    Copyright © 2024 American Chemical Society
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    Abstract

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    A cell-free RNA transcription system had been coupled with electrochemiluminescence (ECL) detection technology for the first time to develop an ascorbic acid (AA, acting as a model target) biosensor. The biosensor is composed of single-stranded DNA (ssDNA) sequences modified with alkynyl and azido groups, respectively, alongside an incomplete gene circuit framework. The addition of target AA and copper ions will cause the linkage of the two ssDNA sequences through a click chemistry reaction. This results in the subsequent reconstruction of a complete gene circuit. The reconstituted gene circuit, in conjunction with the T7 RNA polymerase, drives the transcription of substantial quantities of RNA. ssDNA labeled with ferrocene (Fc) (Fc-DNA) had been immobilized on a tris(2,2′-bipyridyl) ruthenium(II) chloride hexahydrate-doped SiO2 nanoparticle (Ru@SiO2 NPs) modified electrode first. The quenching effect of Fc on Ru@SiO2 causes the low ECL detected. The transcribed RNA sequence assisted double-stranded specific nuclease (DSN) to cut the ssDNA-Fc and the ECL of the system was enhanced. Optimal experimental conditions reveal that the ECL signal exhibits a linear correlation with the logarithmic concentration of AA, spanning a detection range from 100 nM to 1 mM, with a detection limit of 45 nM. This innovative methodology expands the utility of a cell-free RNA transcription system within the realm of biosensing applications.

    Copyright © 2024 American Chemical Society

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    Supporting Information

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    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.analchem.4c04206.

    • Additional experiment details, including the apparatus and the synthesis method of the Ru@SiO2–NH2 NPs; additional figures and table, including the characterization and related optimization of Ru@SiO2 and recoveries of AA in the urine sample (PDF)

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    Analytical Chemistry

    Cite this: Anal. Chem. 2024, 96, 44, 17807–17813
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acs.analchem.4c04206
    Published October 27, 2024
    Copyright © 2024 American Chemical Society
    Request reuse permissions

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