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Active Acetylcholinesterase Immobilization on a Functionalized Silicon Surface.

K Khaldi1, S Sam, A C Gouget-Laemmel2

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This study developed a silicon-based sensor for detecting organophosphorus pesticides by immobilizing active acetylcholinesterase (AChE) enzyme. The functionalized surface achieved a high concentration of active enzyme, showing promise for pesticide detection.

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Area of Science:

  • Biomarker development
  • Surface chemistry
  • Biosensor technology

Background:

  • Organophosphorus (OP) pesticides pose significant environmental and health risks.
  • Acetylcholinesterase (AChE) enzyme activity is a sensitive indicator for OP pesticide exposure.
  • Developing stable and sensitive biosensors for OP detection is crucial.

Purpose of the Study:

  • To create a functionalized silicon surface for immobilizing active acetylcholinesterase (AChE) enzyme.
  • To utilize the immobilized AChE as a potential biomarker for organophosphorus (OP) pesticide detection.
  • To characterize the immobilization process and enzyme activity on the silicon substrate.

Main Methods:

  • Photochemical hydrosilylation to graft a carboxylic acid-terminated monolayer onto a hydrogen-terminated silicon surface.
  • Covalent attachment of AChE via amide bonds using EDC/NHS activation.
  • Characterization using ATR-FTIR and AFM; kinetic studies with in situ real-time IR spectroscopy.
  • Enzymatic activity assessment via colorimetric assays.

Main Results:

  • Successful multistep functionalization of the silicon surface.
  • Quantitative characterization of each modification step.
  • Investigation of enzyme immobilization kinetics.
  • Estimation of surface concentration of active AChE at 1.72 × 10(10) cm(-2).

Conclusions:

  • A robust method for immobilizing active AChE on silicon was established.
  • The functionalized silicon surface shows potential as a platform for OP pesticide detection.
  • The high surface concentration of active enzyme supports its utility as a biomarker.