Preclinical herg assays aim to identify potential inhibitory effects early in drug discovery. Current ich guidelines state that herg assays are mandatory, as a number of clinically successful drugs have had. This method is considered comparison of the predictor herg fp assay to the patch clamp method has previously. This ion channel sometimes simply denoted as herg is. However, there are porbably scenarios where the drug could bind to the herg channel or get into the cell and cause problems. To develop an hts assay for herg using an automated patch clamp. The current strategy is to keep the process as simple as possible and generate manual voltage clamp data only for the herg channel, looking at potency and kinetics of block. Pdf improved throughput of patchxpress herg assay using. Panel c shows a family of herg currents from an experiment described above.
Electrophysiological analysis of mammalian cells expressing herg. Automated patch clamp an overview sciencedirect topics. Animal models experiment systems techinques patchclamp assay from single neuron down to single channel, thanks to the different recording configurations wholecell, perforatedpatch, cellattached. The procedure involves pressing a glass micropipette. Manual patch clamp evaluation of herg channel pharmacology. Technical advances in the automated patch clamp technique and in cell. Since experimental data from a large collection of structurally diverse. Traditionally herg screening is performed with a whole cell voltage patch clamp apparatus. The multiplecompound addition assay has the added benefit of generating cumulative concentrationresponse data similar to those generated in conventional patch clamp assays.
The results presented in table 2 compare single test assumes only one. Automated patchclamp methods for the herg cardiac potassium channel. The herg safety service is performed by our parent company, evotec, and is a cellbased assay which employs the qpatch htx system sophion bioscience as or the syncropatch 384pe nanion. In the automated patchclamp measurements performed here. Invitrogen predictor herg fluorescence polarization assay page 2 of 17 for technical support for this or other drug discovery products, dial 7606037200, option 3, extension 40266 corporate. The cells used for the herg standard patch clamp assay were hek human embryonic kidney 293 cells stably transfected with herg cdna obtained courtesy of m. The herg safety service is performed at our parent company, evotec, and is a cellbased assay which employs the qpatch htx system sophion bioscience as or the syncropatch 384pe nanion.
M a p l i herg k channel binding assays c for cardiac. Covid19 is an emerging, rapidly evolving situation. The impact of automated patch clamping apc on ion channel screening is. Early identification of herg liability in drug discovery programs by.
Inhibition of the herg current causes qt interval prolongation resulting in. Investigation of miscellaneous herg inhibition in large diverse. In addition, we evaluated 19 blinded compounds at 10 m in the. While this article focuses on herg channel recordings using the qpatch sophion as, copenhagen, denmark technology, many of the assay and protocol details given in this article can be. An in vitro electrophysiological assay system, which can assess compound effects and thus show cardiotoxicity including arrhythmia risks of test drugs, is an essential method in the field of.
Whole cell patch clamp recordings from morphologically digitimer ds2a duration. Technologies as automated patch clamp electrophysiology measurements. The report also examines assay technologies preferences used today in the study of ion channels targets today figure 2. Validation of the ionworks barracuda system for herg ion. We also show that the sensitivity of the assay can be improved by adding a. Pdf early identification of herg liability in drug discovery. Automated patchclamp methods for the herg cardiac potassium. Inhibition of herg current at 1 um by patch clamp electrophysiology assay. Currently, the conventional wholecell patch clamp assay is the most reliable. Originally herg blockade was detected by manual electrophysiology, and because of the limitations of xenopus oocytes, patch clamp of mammalian cell lines heterologously expressing herg became the. Figure 2 main assay technologies used to study ion channels targets in drug discovery. Pdf early identification of herg liability in drug. The patch clamp technique is a refinement of the voltage clamp. Solubility assessment and online exposure confirmation in.
Precompound current and postcompound current are measured by. The assay was designed to identify potential herg channel blockers by producing data that accurately correlates with patchclamp electrophysiology studies. Crosssite and crossplatform variability of automated. Admetox, biochemical assays analysis of a new high. Using the predictor herg assay as a model, we demonstrate here the sensitive and fast fluorescence polarization detection capabilities of a new multi. A number of herg assays to assess cardiac safety have been developed e. Cell and tissue culture, electrophysiology, live cell imaging, drug delivery, patch clamp technique, transfection, electrocardiogram, cell proliferation assay. In this assay, choherg cells or hek293herg cells are used to specifically assess the effect of test compounds on the herg channel. These assays include herg ion channel assays employing but not. Biochemical herg assays thermo fisher scientific us. This allows voltageclamp control of the cell and is commonly referred to as a. As an indication of the assay robustness, the pharmacology values obtained from 8 replicates of the experiment were compared for experiment to experiment variation. An in vitro electrophysiological assay system, which can assess compound effects and thus show cardiotoxicity including arrhythmia risks of test drugs, is an. Validation of a multiple compound addition assay for herg.
Technical advances in the automated patch clamp technique and in cell preparations have facilitated employing automated patch clamp assays. High throughput herg assay using the automated patch. Patch clamp recording is an extremely useful technique for investigating the biophysical properties of the ion channels that control neuronal activation. Evaluation of the rubidium efflux assay for preclinical. A patchclamp investigation of hek 293 cells or cho cells expressing herg was performed in an automated way by using a 384wellpatchclamp system. The protocol described in this unit is designed to. Early identification of herg liability in drug discovery.
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