Gradient Microchip Platform for Enzyme Activity and Enzyme Inhibition Assays

Overview

This platform adapts commercially available microfluidic gradient generator chips into a compact system for measuring enzyme activity and inhibition kinetics. By miniaturizing labor-intensive enzyme assays, it reduces reagent and sample volumes, supports parallel testing of multiple conditions, and accelerates workflows used in enzyme characterization and inhibitor screening. The modular design can be configured for sequential or simultaneous testing, making it useful for evaluating enzyme variants, monitoring changes in enzyme activity during storage, and exploring kinetic parameters such as Km and Vmax.

The platform combines fluidics, analytical chemistry, and computation. It is intended to serve industrial R&D groups working with engineered enzymes, enzyme stability monitoring, and inhibitor screening programs.

Technical specifications

How it works:

• Gradient generator microchips split fluid streams into parallel pathways and create controlled gradients of substrate or inhibitor concentrations
• Colorimetric assays are detected using microscope-based imaging; an app-based image analysis tool is planned for quantifying signal
• Computational simulation screens chip designs before fabrication to reduce trial-and-error
• Thin-film-based approaches are explored for placing enzymes within the device
• The inhibition assay can be expanded to multiple inhibitors through multiplexed fluid flow splitters

Key capabilities in development:

• Enzyme activity assays for monitoring different versions of an enzyme and detecting activity changes during storage
• Enzyme inhibition assays using known or proposed inhibitors, with potential to screen several inhibitors concurrently
• Measurement of enzyme kinetic parameters, including Km and Vmax

Technology readiness level

The project is in an early proof-of-concept stage. The research group has demonstrated the feasibility of automated preparation of small-volume substrate solutions using batch-based methods. The proposed work will use readily available chips and microscope-based detection to design, build, and test two enzyme-based assays on a single integrated microfluidic platform. Computational simulations and image analysis tools are planned to complement experimental validation. No commercial deployment has yet occurred; the next step is a pilot collaboration to validate the platform on application-specific enzyme systems.