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Introduction to Solid Supported Membrane Based Electrophysiology
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A Solid Supported Membrane-Based Technology for Electrophysical Screening of B0AT1-Modulating Compounds.

Carolin Gerbeth-Kreul1, Antje Pommereau1, Sven Ruf2

  • 1In Vitro Biology & High-throughput Chemistry, Integrated Drug Discovery, Sanofi-Aventis Deutschland GmbH, Frankfurt, Germany.

SLAS Discovery : Advancing Life Sciences R & D
|May 6, 2021
PubMed
Summary

Solid supported membrane (SSM)-based electrophysiology offers a direct, automated method for screening transporter inhibitors. This study validates SSM electrophysiology for identifying compounds targeting the amino acid transporter B0AT1, reducing false positives.

Keywords:
B0AT1SLC6A19Surfe2rautomated electrophysiologysolid supported membrane-based electrophysiology

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Area of Science:

  • Biophysics
  • Pharmacology
  • Molecular Biology

Background:

  • Classical high-throughput screening (HTS) assays for ionic currents often yield high false-positive rates due to indirect measurements.
  • Electrophysiology-based assays, while direct, are typically complex and not suited for automated screening.
  • There is a need for sensitive, direct, and automated methods for transporter-directed compound screening.

Purpose of the Study:

  • To evaluate the suitability of solid supported membrane (SSM)-based electrophysiology for pharmacological compound identification and optimization.
  • To assess the direct, automated screening capabilities of SSM electrophysiology for transporter proteins.
  • To validate active compounds and identify false positives from primary screens using a direct assay.

Main Methods:

  • Utilized cell-free SSM-based electrophysiological measurements.
  • Focused on the electrogenic amino acid transporter B0AT1 (SLC6A19).
  • Electrophysiological characterization, inhibition assays with a tool compound, and assessment of assay robustness and reproducibility were performed.

Main Results:

  • Leucine-induced currents were specific to B0AT1, confirming target engagement.
  • B0AT1 activity was successfully inhibited by an in-house tool compound.
  • The SSM assay demonstrated robustness, reliability, and the ability to validate primary screening hits while identifying false positives.

Conclusions:

  • SSM-based electrophysiology is a suitable direct method for rapid and automated identification of small molecules inhibiting B0AT1 activity.
  • This technique offers an orthogonal approach to validate compounds identified through conventional HTS.
  • SSM electrophysiology enhances the efficiency and accuracy of drug discovery for transporter targets.