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A new method uses fluorescent nanoparticles and MnO2 nanosheets to quantify acetylcholinesterase (AChE) activity. This single-particle enumeration technique offers sensitive detection of AChE and inhibitors in biological samples.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Nanotechnology

Background:

  • Acetylcholinesterase (AChE) is crucial for nerve function and linked to Alzheimer's disease.
  • Accurate quantification of AChE activity is essential for disease diagnosis and drug development.
  • Existing methods for AChE detection often lack sensitivity or require complex procedures.

Purpose of the Study:

  • To develop a novel, label-free single-particle enumeration (SPE) method for quantifying acetylcholinesterase (AChE) activity.
  • To utilize fluorescence resonance energy transfer (FRET) between polymer nanoparticles and MnO2 nanosheets for sensitive detection.
  • To demonstrate the method's applicability in biological samples and for inhibitor screening.

Main Methods:

  • Ache-based hydrolysis of acetylthiocholine (ATCh) reduces MnO2 nanosheets, restoring fluorescence from conjugated polymer nanoparticles (FCPNPs).
  • Single-particle enumeration (SPE) quantifies AChE activity by counting restored fluorescent particles.
  • The method was validated using human serum samples and for detecting pesticide inhibitors.

Main Results:

  • The SPE method achieved a linear detection range of 5–1600 μU mL-1 for AChE with a limit-of-detection (LOD) of 1.02 μU mL-1.
  • Satisfactory recovery efficiencies (91.0%-103.0%) were obtained in human serum samples.
  • The method demonstrated high sensitivity for detecting the AChE inhibitor carbaryl, with an LOD of 1.12 pg mL-1.

Conclusions:

  • The label-free SPE method provides a sensitive and accurate platform for quantifying AChE activity.
  • This approach is suitable for detecting AChE and its inhibitors in complex biological matrices.
  • The developed technique holds promise for future applications in diagnostics and drug screening.