Qube as a tool for assay optimization of CiPA cells and protocols by using multiple IC, EC solutions and hERG, Nav1.5 and Cav1.2 on the same QChip

著者: Melanie Schupp, Anders Lindqvist, Pia Bladsgaard Andersen, Weifeng Yu

The Comprehensive in vitro Proarrhythmia Assay (CiPA) proposal provides an attractive perspective for increasing the efficacy of the drug development process. A detailed electrophysiological analysis of Nav1.5 (peak and late currents), Kv4.3 (Ito), hERG (IKr), KvLQT1/minK (IKs), Cav1.2 and Kir2.1 (IK1) upon addition of a potential drug is a major part of assessing the drug’s proarrhythmic risk. Gaining a better understanding of the complex relationship between QT-elongation and the occurence of Torsades des Pointes (TdP), potentially enables compounds with properties that today are considered as problematic to be further developed. High throughput screening (HTS) supports this quest and is furthermore of great importance for discovering pharmacologically active substances and understanding ion channels.

Qube is a giga-seal automated patch clamp (APC) instrument, providing 384 amplifiers for the consumable QChip 384 with its integrated electrodes. The QChip has built-in microfluidic flow channels that ensure a fast and complete exchange of liquid for reliable measurements on ligand-gated ion channels and sequential additions to the same site. Here, four different cell lines expressing the cardiac ion channels Nav1.5, Cav1.2, Kv1.5 and hERG, were transferred from a cell clone cell transfer plate (ccCTP) onto the same QChip. A range of pharmacological substances and voltage step protocols were applied to address the suitability of Qube for measuring different cell populations in parallel.