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Detection of Pyrophosphate and Alkaline Phosphatase Activity Based on PolyT Single Stranded DNA - Copper Nanoclusters

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Abstract

The determination of pyrophosphate and alkaline phosphatase activity plays a significant role in medical diagnosis. In this work, a label-free “ON-OFF-ON” fluorescence strategy is developed for the analysis of pyrophosphate and alkaline phosphatase activity. Using PolyT single strand DNA as templates to synthesize fluorescent copper nanoparticles, the coordination effect of pyrophosphoric acid on Cu2+ inhibited the generation of fluorescence. Afterwards, the addition of alkaline phosphatase into hydrolyze pyrophosphoric acid resulted in the release of Cu2+, whereby the fluorescence intensity could be recovered. Thereupon enhanced-sensitivity for alkaline phosphatase was obtained (0.1 mU/L), much better than previously reported methods. Meanwhile, it could be performed directly in homogeneous solution, which was very close to the actual activity level of alkaline phosphatase under physiological conditions. Likewise, satisfactory results were also obtained in specificity assessment, which demonstrated its potential application in clinical diagnosis. Notably, a new, sensitive, low-cost, short-time, and high-sensitivity platform for alkaline phosphatase detection was constructed, and the design of biosensor using DNA-templated Copper nanoclusters (CuNCs) was instructed in this study.

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Data Availability

All data generated or analysed during this study are included in this published article.

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Funding

This work was supported in part by the Hunan Provincial Natural Science Foundation of China (2021JJ31138), Scientific Research Fund of Hunan Provincial Education Department (20A520), Training Program for Excellent Young Innovators of Changsha (kq2106053) and Hunan Province graduate research innovation project (CX20210889), Hunan Province Student Innovation and Entrepreneurship Training Program.

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Authors and Affiliations

  1. Hunan Province Key Laboratory for Forestry Biotechnology, College of Life Science and technology, Central South University of Forestry and Technology, 410004, Changsha, Hunan Province, China

    Liu Yao, Xin Li, Hong Li, Zhibin Liao, Chuchu Xie, Yaohui Wu & Yonghong Wang

  2. International Education Institute, Hunan University of Chinese Medicine, 410208, Changsha, Hunan Province, China

    Ge Ning

Authors
  1. Liu Yao
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  2. Xin Li
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  3. Hong Li
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  4. Zhibin Liao
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  5. Chuchu Xie
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  6. Ge Ning
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  7. Yaohui Wu
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  8. Yonghong Wang
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Liu Yao, Xin Li, Hong Li, Zhibin Liao, Chuchu Xie, Ge Ning, Yaohui Wu, Yonghong Wang. The first draft of the manuscript was written by Liu Yao and all authors commented on previous versions of the manuscript, Writing - review and editing by Yonghong Wang, Ge Ning, All authors read and approved the final manuscript, Liu Yao and Xin Li are co-first authors.

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Correspondence to Yonghong Wang.

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This is an observational study. The XYZ Research Ethics Committee has confirmed that no ethical approval is required.

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Yao, L., Li, X., Li, H. et al. Detection of Pyrophosphate and Alkaline Phosphatase Activity Based on PolyT Single Stranded DNA - Copper Nanoclusters. J Fluoresc 32, 1949–1957 (2022). https://doi.org/10.1007/s10895-022-02984-5

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  • DOI: https://doi.org/10.1007/s10895-022-02984-5

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