How to extend range linearity in enzyme inhibition-based biosensing assays

Area of Science:

• Analytical Chemistry
• Biochemistry
• Enzyme Kinetics

Background:

• Enzyme inhibition bioassays are valuable analytical tools but often suffer from narrow linear ranges.
• Conventional methods like chromatography and mass spectrometry can be costly and time-consuming.

Purpose of the Study:

• To develop a novel graphical method to extend the linear range of enzyme inhibition bioassays.
• To provide a more accurate and efficient method for inhibitor analysis.

Main Methods:

• Utilized the integrated Michaelis-Menten equation for a novel graphical analysis.
• Applied the method to quantify fluoride using acetylcholinesterase inhibition.
• Demonstrated applicability with catalase enzyme inhibition by cyanide.
• Investigated fixed substrate conversion for inhibitor determination.

Main Results:

• Extended the linear range for fluoride quantification up to 5 mM, significantly improving upon the conventional 0.6 mM limit.
• Successfully applied the method to catalase-cyanide inhibition.
• Achieved wide linear range determination with high precision and speed using fixed substrate conversion.

Conclusions:

• The novel graphical method effectively extends the linear range of enzyme inhibition bioassays for various reversible inhibitions.
• This approach offers advantages such as reduced reagent consumption, waste generation, and measurement time.
• It provides a cost-effective and efficient alternative for inhibitor analysis.