Black phosphorus nanosheets based sensitive protease detection and inhibitor screening

Area of Science:

• Biochemistry
• Materials Science
• Analytical Chemistry

Background:

• Proteases are crucial digestive enzymes involved in numerous physiological processes and diseases.
• Developing sensitive and selective methods for protease detection is vital for understanding biological functions and disease mechanisms.
• Black phosphorus nanosheets (BPs) are emerging two-dimensional (2D) materials with unique properties suitable for biosensing applications.

Purpose of the Study:

• To develop a sensitive and label-free fluorometric method for detecting protease activity.
• To utilize black phosphorus nanosheets (BPs) as a platform for protease detection.
• To establish a method for high-throughput screening of protease inhibitors.

Main Methods:

• A fluorescence-based assay was designed utilizing a perylene probe (probe 1) and black phosphorus nanosheets (BPs).
• BPs were shown to quench the fluorescence of probe 1 through electrostatic adsorption.
• Histone was employed to modulate the interaction between BPs and the probe, allowing protease-regulated fluorescence changes to be monitored.

Main Results:

• The proposed method demonstrated high sensitivity and selectivity for protease detection.
• The fluorescence intensity of probe 1 was directly correlated with protease activity.
• The assay successfully detected as low as 1 ng/mL of trypsin.
• The method proved effective for label-free detection and inhibitor screening.

Conclusions:

• The developed black phosphorus nanosheets (BPs)-based fluorometric method offers a sensitive, selective, and label-free approach for protease detection.
• This technique provides a valuable tool for monitoring protease activity in biological samples.
• The method holds promise for applications in enzyme-linked assays and drug discovery for protease-related diseases.