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Nanozyme-Enzyme Cascade Reaction-Enhanced Ratiometric Fluorescence Immunosensing Platform for Sensitive and Accurate

Liang Luo1,2, Chaochao Chen3, Jincheng Xiong4

  • 1National Key Laboratory of Veterinary Public Health and Safety, Beijing Key Laboratory of Detection Technology for Animal Derived Food Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China.

Journal of Agricultural and Food Chemistry
|November 13, 2024
PubMed
Summary

A novel ratiometric fluorescence (RF) sensing platform using nanozyme-enzyme cascade reactions offers enhanced sensitivity and stability for detecting trace analytes like ractopamine (RAC), overcoming limitations of traditional ELISAs.

Keywords:
MnO2 nanosheet nanozymecascade reactionractopamineratiometric fluorescencesensing platform

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Materials Science

Background:

  • Traditional enzyme-linked immunosorbent assays (ELISA) suffer from poor sensitivity and stability due to perishable enzymes, limiting trace analyte detection.
  • Developing robust and sensitive detection methods is crucial for various applications, especially in challenging environments.

Purpose of the Study:

  • To develop a novel ratiometric fluorescence (RF) sensing platform for enhanced detection of analytes.
  • To create a nanozyme-enzyme cascade reaction for signal amplification.
  • To establish a sensitive and stable RF ELISA for ractopamine (RAC) detection.

Main Methods:

  • A ratiometric fluorescence (RF) sensing platform was constructed using manganese dioxide nanosheets (MnO2 NSs) as an oxidase-like nanozyme and alkaline phosphatase (ALP).
  • The platform utilizes the catalytic activity of MnO2 NSs on Amplex Red (AR) and their quenching effect on carbon dots (CDs).
  • An enzymatic cascade involving ALP, ascorbic acid 2-phosphate (AAP), and ascorbic acid (AA) was employed to regulate the fluorescence signals.

Main Results:

  • The RF sensing platform achieved a superior limit of detection (LOD) of 0.037 mU mL-1 for ALP activity.
  • A sensitive and accurate RF ELISA for ractopamine (RAC) detection was established with an LOD of 0.029 ng mL-1.
  • The RF ELISA demonstrated a 5.5-fold increase in sensitivity compared to traditional colorimetric ELISA.

Conclusions:

  • The proposed RF sensing platform, based on a robust nanozyme-triggered enzymatic cascade, offers a powerful and universal approach for trace target detection.
  • This method overcomes the limitations of traditional immunoassays, providing enhanced sensitivity and stability.
  • The developed RF sensing platform holds significant potential for detecting ractopamine and other trace analytes in various applications.