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Updated: Apr 14, 2026

High-throughput Screening for Small-molecule Modulators of Inward Rectifier Potassium Channels

High-throughput Screening for Small-molecule Modulators of Inward Rectifier Potassium Channels

Published on: January 27, 2013

POLARISED views and FRETting about probe modulation assays: Learning from High Throughput Screening.

Alice Lanne¹, Catherine Bardelle², Gareth Davies²

¹Hit Discovery, Discovery Sciences, R&D, AstraZeneca, Cambridge, UK.

SLAS Discovery : Advancing Life Sciences R & D

|April 20, 2024

Summary

Fluorescent probe modulation assays effectively identify compounds that interact with target proteins. These high-throughput screening methods, utilizing fluorescence, are crucial for drug discovery and identifying novel molecular glues.

Keywords:

Fluorescence polarisation High throughput screening Probe modulation Time resolved Förster resonance energy transfer

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Area of Science:

Biochemistry
Pharmacology
Drug Discovery

Background:

Fluorescent probe modulation assays are standard for assessing ligand binding.
These assays detect compounds displacing or stabilizing fluorescently labeled ligands.
They are applicable to both orthosteric and allosteric sites.

Purpose of the Study:

To present the application of fluorescent probe modulation assays in high-throughput screening.
To detail assay development, screening timelines, and hit-finding strategies.
To report on the outcomes of eight high-throughput screens conducted in 2023.

Main Methods:

Utilizing fluorescently labeled tool ligands and test compounds.
Employing streptavidin labeled terbium as a donor with various acceptor fluorophores.
Implementing orthogonal methods to validate hits and identify artefacts.

Main Results:

Eight high-throughput screens were successfully executed.
Analysis included hit rates and the number of compounds with IC50 values below 30 µM.
Key learnings from assay development, execution, and analysis were documented.

Conclusions:

Fluorescent probe modulation assays are versatile tools for drug discovery.
These fluorescence-based methods are valuable for identifying modulators and molecular glues.
The presented screens demonstrate the utility and efficiency of these assays in a high-throughput screening context.