Borinic Acid-Based Fluorogenic Probes as an Alternative to the Amplex Red Assay for Real-Time H₂O₂ Monitoring in Live...

Area of Science:

• Biochemistry
• Chemical Biology
• Cell Biology

Background:

• Hydrogen peroxide (H2O2) is a vital reactive oxygen species (ROS) regulating physiological and pathological processes.
• Dysregulated H2O2 production causes oxidative stress linked to aging, cancer, and neurodegenerative diseases.
• Stable H2O2 requires probes with high selectivity, sensitivity, and rapid response for accurate monitoring.

Purpose of the Study:

• Develop novel fluorogenic probes for sensitive and real-time H2O2 detection.
• Overcome limitations of existing boronic acid-based probes, particularly their reactivity.
• Enable monitoring of H2O2 dynamics in cellular environments, both intra- and extracellularly.

Main Methods:

• Design and synthesis of new borinic acid-based fluorogenic probes.
• Utilizing a hemicyanine scaffold functionalized with a borinic acid trigger.
• Evaluating probe kinetics and performance in cellular models.

Main Results:

• Borinic acid probes exhibited superior reaction kinetics compared to boronic acid counterparts.
• Probes enabled efficient, real-time monitoring of H2O2 in cellular models.
• Enzyme-free probe kinetics matched the Amplex UltraRed/horseradish peroxidase (HRP) assay.

Conclusions:

• The developed borinic acid probes represent a significant advancement for H2O2 detection.
• These probes offer a versatile and sensitive tool for studying H2O2-mediated cell signaling.
• The superior reactivity facilitates real-time monitoring crucial for understanding H2O2's pathophysiological roles.