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Thread-based microfluidic chips as a platform to assess acetylcholinesterase activity.

Ariana Gonzalez1, Michelle Gaines1, Frank A Gomez1

  • 1Department of Chemistry and Biochemistry, California State University, Los Angeles, Los Angeles, CA, USA.

Electrophoresis
|January 7, 2017
PubMed
Summary

A novel microfluidic thread-based analytical device (μTAD) enables colorimetric analysis of acetylcholinesterase (AChE) activity. This easy-to-use device shows potential for point-of-care diagnostics in resource-limited settings.

Keywords:
AcetylcholinesteraseAssayPaper microfluidics

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Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Materials Science

Background:

  • Acetylcholinesterase (AChE) is a crucial enzyme in neurotransmission.
  • Accurate and accessible methods for AChE activity assessment are needed, particularly for point-of-care applications.
  • Microfluidic devices offer miniaturized platforms for biochemical assays.

Purpose of the Study:

  • To develop and characterize a microfluidic thread-based analytical device (μTAD) for colorimetric assessment of acetylcholinesterase (AChE) activity.
  • To demonstrate the device's capability for quantitative analysis and inhibitor detection.
  • To explore the potential of μTADs for resource-challenged diagnostic settings.

Main Methods:

  • Fabrication of a trifurcated nylon thread microfluidic device with integrated analysis sites.
  • Colorimetric detection of AChE activity using 5,5'-Dithiobis-(2-nitrobenzoic acid) (DTNB) and acetylthiocholine iodide (ATC) substrate.
  • Quantitative analysis of color intensity and determination of IC50 values for inhibitors.

Main Results:

  • The μTAD successfully detected AChE activity via a colorimetric reaction producing a yellow anion (TNB^2-).
  • A linear relationship was observed between inverse yellow mean intensity and substrate concentration.
  • An IC50 value of 1.74 nM was determined for the inhibitor neostigmine bromide (NB).
  • The device design allows for multiplexed, triplicate data collection.

Conclusions:

  • The developed μTAD is a simple, effective, and quantitative platform for assessing AChE activity.
  • The device's multiplexing capability and ease of use are advantageous for data reliability and accessibility.
  • μTADs hold significant promise for development into point-of-care diagnostic tools, especially in resource-limited environments.