Detection of protease activities by flash chronopotentiometry using a reversible polycation-sensitive polymeric...

Area of Science:

• Electrochemistry
• Biochemistry
• Analytical Chemistry

Background:

• Protease activity is crucial in biological processes.
• Existing methods for protease detection can be time-consuming or lack sensitivity.
• A need exists for rapid and sensitive protease detection techniques.

Purpose of the Study:

• To develop a novel electrochemical method for rapid and sensitive protease activity detection.
• To establish flash chronopotentiometry (FCP) as a viable technique for enzyme activity monitoring.
• To correlate changes in transition time to specific protease activities.

Main Methods:

• Utilized flash chronopotentiometry (FCP) with a polycation-selective polymer membrane electrode.
• Applied a current pulse to induce peptide flux into the electrode membrane.
• Monitored cell potential (EMF) continuously to determine transition time (τ).
• Correlated changes in transition time (τ) to protease-induced cleavage of polypeptides.

Main Results:

• Demonstrated a direct relationship between the square root of transition time (τ) and polypeptide concentration.
• Observed a decrease in transition time (τ) upon protease activity due to peptide cleavage.
• Successfully detected the activity of trypsin and α-chymotrypsin using protamine and synthetic peptides.
• Showcased the sensitivity and rapidity of the FCP method for protease detection.

Conclusions:

• Flash chronopotentiometry (FCP) provides a rapid and sensitive method for detecting protease activity.
• The technique is based on the electrochemical detection of protease-induced changes in polypeptide concentration.
• FCP offers a promising new tool for biochemical analysis and enzyme activity monitoring.