Fluorescence biosensing micropatterned surfaces based on immobilized human acetylcholinesterase

Area of Science:

• Biochemistry
• Materials Science
• Nanotechnology

Background:

• Human acetylcholinesterase (AChE) is a key target for Alzheimer's disease (AD) drug discovery.
• Developing sensitive biosensing surfaces is crucial for evaluating AChE inhibitors.

Purpose of the Study:

• To optimize the material structure and chemistry of a novel AChE-based fluorescence sensing surface.
• To identify the ideal substrate for maximizing fluorescence signal and chemical stability.

Main Methods:

• Fabrication of multilayered silicon wafers with patterned silicon oxide, platinum, and titanium dioxide layers.
• Labeling with HiLyte Fluor 555 and evaluation using confocal laser-scanning microscopy (CLSM).
• Immobilization of human AChE and characterization using atomic-force microscopy (AFM) and CLSM.

Main Results:

• Wafers with 100 nm thick silicon oxide features yielded the highest signal-to-background ratio.
• Immobilized AChE retained its enzymatic activity.
• AChE was homogeneously and selectively distributed on silicon oxide microstructures.

Conclusions:

• The optimized SiO(2)/Pt/Ti/Si wafer design with 100 nm features is suitable for AChE immobilization.
• The developed surface enables efficient fluorescence emission for AChE detection.
• This biosensing surface shows promise for AD drug discovery applications.