Abstract
Recently, a couple of giga-seal automated patch clamp (APC) platforms in a standard 384-well plate format have been introduced. These APC instruments can deliver higher throughput measurements with high-quality data for ion channel drug discovery. In this study, we developed an automated electrophysiology assay for the voltage-gated potassium channel, Kv1.3 as a test case on the Nanion SyncroPatch 384PE, one of the latest APC platforms. We achieved a high cell-catch rate (98% wells/plate, defined by membrane resistance >16 MΩ/cell) by screening plate types (number of holes and resistance), and obtained a stable seal (87% wells/plate, defined by initial/final seal resistance >500 MΩ/cell) by optimizing recording solutions. We generated data at an overall success rate (defined by initial peak current >300 pA/cell, initial/final seal resistance >500 MΩ/cell, and current stability <±7% change/min) of 79% on average. Using the same platform, we validated a dose-response assay using clotrimazole, a potent Kv1.3 inhibitor. The assay was robust with Z' factor 0.52 and the success rate 75%. Altogether, our results demonstrated that the SyncroPatch represents a reliable platform for voltage-gated potassium channel assays.
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