Droplet-Based Microfluidics Methods for Detecting Enzyme Inhibitors

Abstract

Sub-nanoliter droplets produced in microfluidic devices have gained an enormous importance for performing all kinds of biochemical assays. One of the main reasons is that the amounts of reagents employed can be reduced in approximately five orders of magnitude compared to conventional microplate assays. In this chapter, we describe how to carry out the design, fabrication, and operation of a microfluidic device that allows performing enzyme kinetics and enzyme inhibition assays in droplets. This procedure can be used effectively to screen a small size library of compounds. Then, we describe how to use this droplet microfluidic setup to screen for potential inhibitor compounds eluted from a coupled high-performance liquid chromatography (HPLC) system that separates crude natural extracts.

Keywords: Droplet microfluidics; Droplet-based assays; Drug discovery; Enzymatic assays; Enzyme inhibition; Enzyme kinetics; High-throughput screening; Microdroplets; Microfluidic device; Natural products.