Label-free measurements of reaction kinetics using a droplet-based optofluidic device

J Lab Autom. 2015 Feb;20(1):17-24. doi: 10.1177/2211068214549625. Epub 2014 Sep 23.

Abstract

Label-free measurements of the reaction kinetics of a small sample volume are essential for efficient drug discovery, requiring methods and systems that are rapid, accurate, and cost-effective. Herein, we present an integrated optofluidic system for label-free characterization of reactions in a nanoliter reagent volume. This system contains a droplet-based microfluidic sampling section and an optical fiber-based spectroscopy detection section. By manipulating droplets containing reagents at certain concentrations at different times, quantifiable measurements via absorption spectroscopy can be made in a simple, sensitive, and high-throughput manner. We have demonstrated our system's capability by performing potency (IC50) assays of an inhibitor in a TEM-1 β-lactamase (enzyme) and nitrocefin (substrate) system. This integrated platform can potentially provide an automated, label-free, and low-cost method for many other assays of reaction kinetics.

Keywords: absorption spectroscopy; droplet; label-free detection; microfluidics; optofluidics; reaction kinetics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Drug Discovery / methods*
  • Drug Evaluation, Preclinical / methods*
  • Enzyme Inhibitors / metabolism
  • High-Throughput Screening Assays / instrumentation
  • High-Throughput Screening Assays / methods
  • Inhibitory Concentration 50
  • Kinetics
  • Microfluidics / instrumentation*
  • Microfluidics / methods*
  • Spectrum Analysis / instrumentation*
  • Spectrum Analysis / methods*
  • beta-Lactamases / metabolism

Substances

  • Enzyme Inhibitors
  • beta-Lactamases
  • beta-lactamase TEM-1