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Related Concept Videos

Patch Clamp01:18

Patch Clamp

6.1K
Many fundamental cell functions such as muscle contraction and nerve transmission rely on the electrical signals produced by the movement of positively and negatively charged ions across the cell membrane. One competent method to record current flowing across the whole cell or single ion channel is the patch-clamp technique.
In this method, a glass micropipette containing electrolyte solution is tightly sealed against a small portion of the cell membrane. As a result, a patch of the cell...
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One-channel Cell-attached Patch-clamp Recording
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Screening Assay Protocols Targeting the Nav1.7 Channel Using Qube High-Throughput Automated Patch-Clamp System.

Binbin Qian1, Sung Hoon Park1, Weifeng Yu1

  • 1Sophion Bioscience A/S, Baltorpvej, Ballerup, Denmark.

Current Protocols in Pharmacology
|April 14, 2020
PubMed
Summary

This study details protocols for Nav1.7 screening using automated patch-clamp systems. New adaptive online methods improve Vhalf estimation and enable state- and use-dependent inhibition assessments.

Keywords:
Nav1.7Qubeautomated patch clamphigh-throughput screeningvoltage-gated sodium channels

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Area of Science:

  • Neuroscience
  • Pharmacology
  • Biophysics

Background:

  • Nav1.7 ion channels are crucial drug targets.
  • Automated patch-clamp systems offer high-throughput screening capabilities.
  • Accurate characterization of channel gating is essential for drug development.

Purpose of the Study:

  • To describe protocols for Nav1.7 screening on a Qube automated patch-clamp system.
  • To introduce and validate novel adaptive online methods for Vhalf estimation.
  • To establish state- and use-dependent protocols for assessing channel blockers.

Main Methods:

  • Utilized a Qube automated patch-clamp system.
  • Implemented three distinct screening protocols: steady-state inactivation, state-dependent, and use-dependent inhibition.
  • Employed traditional offline calculation and a new adaptive online estimation for Vhalf.
  • Performed concentration-response curve (CRC) analysis for reference blockers.

Main Results:

  • Successfully established protocols for Nav1.7 screening.
  • Demonstrated the utility of adaptive online Vhalf estimation for improved efficiency.
  • Validated state- and use-dependent inhibition assays using a reference blocker.
  • The reference blocker showed potent state- and use-dependent inhibition of Nav1.7.

Conclusions:

  • The described protocols provide a robust framework for Nav1.7 screening.
  • Adaptive online Vhalf measurement enhances the efficiency of electrophysiological assays.
  • State- and use-dependent protocols are effective for characterizing Nav1.7 channel blockers.
  • These methods facilitate the discovery of novel therapeutics targeting Nav1.7.