High-throughput electrophysiological assays for voltage gated ion channels using SyncroPatch 768PE

Area of Science:

• Electrophysiology
• Pharmacology
• Biophysics

Background:

• Ion channels are crucial physiological regulators and drug targets.
• Patch clamp electrophysiology is the gold standard for ion channel studies but suffers from low throughput.
• Automated patch clamp (APC) platforms aim to increase throughput but vary in performance.

Purpose of the Study:

• To evaluate the SyncroPatch 768PE, a new APC platform, for high-throughput ion channel drug discovery.
• To develop and validate a robust APC assay for the voltage-gated sodium channel Nav1.7.
• To demonstrate the platform's capability in recording endogenous ion channels from primary cells.

Main Methods:

• Utilized the SyncroPatch 768PE for parallel recordings.
• Optimized cell patching parameters and a two-step voltage protocol for Nav1.7 assay development.
• Tested reference compounds for Nav1.7 IC50 and screened 10,000 compounds.
• Recorded endogenous Kv1.3 channels in primary T cells.

Main Results:

• The Nav1.7 assay demonstrated high consistency with manual patch clamp (R > 0.9).
• Achieved a 79% success rate in a pilot screen of 10,000 compounds.
• Demonstrated robust assay performance with ~6,000 data points daily throughput and a Z' factor of 0.72.
• Successfully recorded endogenous Kv1.3 channels in primary T cells.

Conclusions:

• The SyncroPatch 768PE is a powerful, high-throughput platform for ion channel research.
• The developed APC assay is suitable for Nav1.7 drug discovery.
• The platform enables characterization of endogenous ion channels in primary cells, advancing drug discovery efforts.