Issue 19, 2013

Microfluidics platform for single-shot dose–response analysis of chloride channel-modulating compounds

Abstract

We previously developed cell-based kinetics assays of chloride channel modulators utilizing genetically encoded yellow fluorescent proteins. Fluorescence platereader-based high-throughput screens yielded small-molecule activators and inhibitors of the cAMP-activated chloride channel CFTR and calcium-activated chloride channels, including TMEM16A. Here, we report a microfluidics platform for single-shot determination of concentration–activity relations in which a 1.5 × 1.5 mm square area of adherent cultured cells is exposed for 5–10 min to a pseudo-logarithmic gradient of test compound generated by iterative, two-component channel mixing. Cell fluorescence is imaged following perfusion with an iodide-containing solution to give iodide influx rate at each location in the image field, thus quantifying modulator effects over a wide range of concentrations in a single measurement. IC50 determined for CFTR and TMEM16A activators and inhibitors by single-shot microfluidics were in agreement with conventional plate reader measurements. The microfluidics approach developed here may accelerate the discovery and characterization of chloride channel-targeted drugs.

Graphical abstract: Microfluidics platform for single-shot dose–response analysis of chloride channel-modulating compounds

Supplementary files

Article information

Article type
Paper
Submitted
10 Jul 2013
Accepted
17 Jul 2013
First published
24 Jul 2013

Lab Chip, 2013,13, 3862-3867

Microfluidics platform for single-shot dose–response analysis of chloride channel-modulating compounds

B. Jin, E. Ko, W. Namkung and A. S. Verkman, Lab Chip, 2013, 13, 3862 DOI: 10.1039/C3LC50821H

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