Ambient temperature CiPA dynamic hERG protocol dataset recorded on the SyncroPatch 384SE

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. The application of this approach at scale 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 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% overall, or as high as 50.4% when only appropriate sweep lengths were considered for drugs with faster kinetics. 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 the modifications to protocol design, including sweep length, number of repetitions, and leak correction recommended in this study, this protocol can be applied on APC to acquire data comparable to manual patch clamp.

Description

High-throughput patch clamp electrophysiology dataset acquired on Nanion SyncroPatch 384PE platform describing the potency and kinetics of drug block of hERG channels stably expressed in CHO cells. The voltage protocols used were 10 or 40 s 0mV steps with a 15s, -80mV inter-pulse intervals, repeated 5 or 10 times in the absence (control) and presence of drug. Drug was added and cells clamped at -80 mV for 4 minutes prior to recording to ensure drug equilibration prior to recording the onset (kinetics) of drug block. This dataset accompanies the paper titled 'High throughput measurement of hERG drug block kinetics using the CiPA dynamic protocol' by Monique J. Windley, Jessica Farr, Clifford TeBay, Jamie I. Vandenberg and Adam P. Hill. 

File information

Raw and leak corrected files for individual plates run on the SyncroPatch 384PE

Folder naming convention: Plate identifier (6 digit date_user initials_plate number)_type of data (leak uncorrected raw / leak corrected).

Raw data

File naming convention: Plate identifier (6 digit date_user initials_plate number)_well number

Each raw data folder also contains a file named QC_plate identifier no. which contains information used for cell/patch and hERG current quality controls including seal resistance, series resistance, capacitance, baseline (average current at -80mV), average (average leak uncorrected current during last 100 ms of 0mV step). QC details are reported for each sweep along with the drug and concentration, with external and 0 M reported for control sweeps.

Full SyncroPatch plates were not used for all runs, only applicable wells were included where  partial plates were run (see plates 240719_MW1-3, 170719_MW1-3, 180719_MW1 and  291119_MW1)

Leak corrected data

File naming convention: well number_drug_concentration in M_leak_correction

Refer to folder for plate identifier number.

Each folder containing leak corrected data also contains a file labelled 'percent block' which contains percentage block measurements for each leak correction file in the same folder as identified by the well number.

Only files passing the first pass QC were leak corrected (>125 MOhm seal, <80 MOhm series, 10-200 pA capacitance, >-300 pA baseline, >50 pA hERG current size and <20% change in peak control current)