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    CardioExcyte 96 SOL:用光遗传的手段起搏心肌细胞

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2022 - Translating the Measurement of Herg Kinetics and Drug Block for Cipa to a High Throughput Platform

icon sp96 SyncroPatch 384PE (a predecessor model of the SyncroPatch 384 instrument) pre-print publication in SSRN (2022)

Authors:
Windley M.J., Farra J., Vandenberg J.I., Hill A.P.

Journal:
SSRN (2022) doi:10.2139/ssrn.4065593


Abstract: 

The Comprehensive in vitro Proarrhythmic Assay (CiPA) has promoted use of in silico models of drug effects on cardiac repolarization to improve proarrhythmic risk prediction. These models contain a pharmacodynamic component describing drug binding to hERG channels that required in vitro data for kinetics of block, in addition to potency, to constrain them. To date, development and validation has been undertaken using data from manual patch-clamp. To enable the application of this approach at scale this requires the development of a high-throughput, automated patch-clamp (APC) implementation. Here, we present a comprehensive analysis of the implementation of the Milnes, or
CiPA dynamic protocol, on an APC platform, including automated quality control and data analysis. Kinetics and potency of block were assessed for bepridil, cisapride, terfenadine and verapamil with data retention/QC pass rate of 21.8%. The variability in IC50 and kinetics between manual and APC was comparable to that seen between sites/platforms in previous APC studies of potency. Whilst the experimental success is less than observed in screens of potency alone, it is still significantly greater than manual patch. With appropriate consideration of protocol design, including sweep length, number of repetitions, and leak correction, this protocol can be applied on APC to acquire data comparable to manual patch clamp.


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