Label-Free and Bioluminescence-Based Nano-Biosensor for ATP Detection

Area of Science:

• Biomedical Engineering
• Nanotechnology
• Biochemistry

Background:

• Cell viability assays often rely on adenosine-5'-triphosphate (ATP) concentration, necessitating sensitive and cost-effective detection methods.
• Gold nanoparticles (AuNPs) offer excellent biocompatibility for enzyme immobilization, crucial for developing biosensors.
• Existing ATP detection methods can be limited by signal stability and detection sensitivity.

Purpose of the Study:

• To design and evaluate a novel, low-cost, and user-friendly adenosine-5'-triphosphate (ATP) biosensor.
• To investigate the impact of various gold-based nanoparticles on luciferase activity and signal stability.
• To develop an ATP biosensor utilizing a DNA aptamer and firefly luciferase immobilized on gold nanoparticles.

Main Methods:

• Investigated the effect of different metal nanoparticles (gold, silver, core-shell) and nanostructures (nanorods, nanoclusters) on luciferase activity.
• Utilized a bioluminescence assay to quantify enzyme activity and signal stability in the presence of nanoparticles.
• Developed a biosensor using a 27-base DNA aptamer for ATP detection with firefly luciferase and gold nanoparticles.

Main Results:

• Metal nanoparticles enhanced luciferase activity by 3-5 times compared to the free enzyme.
• Signal stability improved significantly in the presence of nanoparticles, remaining reliable for up to 6 hours.
• The developed ATP biosensor demonstrated a detection limit of 5 µM for ATP, with increased bioluminescence intensity up to 600 µM.

Conclusions:

• Gold-based nanoparticles enhance luciferase activity and signal stability, making them suitable for ATP biosensor development.
• The DNA aptamer-functionalized gold nanoparticle biosensor offers improved sensitivity and stability for ATP detection.
• This approach provides a promising platform for low-cost and efficient cell viability assessment through ATP measurement.