Category: Assay Development and Screening
High Throughput Screening for the European Lead Factory (ELF) is performed at Pivot Park Screening Centre (PPSC). Over 55 HTS campaigns have been performed in the past 4 years on a compound library that has grown from 320.000 to 450.000 compounds. The screens cover a wide range of target classes including more demanding cellular targets like ion channels and protein-protein interactions.
As an example of one of the ELF screening campaigns we present the results of a High Throughput Screening to identify selective agonists for an ion channel using a Fluoremetic Imaging Plate Reader (FLIPR). In this project, the assay was based on stable cell lines expressing the ion channel. The assay was successfully miniaturized to 1536 well format using a standard membrane potential dye (Molecular Devices). The BlueWasher (BlueCatBio ) was evaluated and shown to give more reproducible and efficient removal of growth medium leading to higher Z’. Therefore, the BlueWasher was integrated into the robot system to allow for efficient, automated screening. The PPSC Robustness set, consisting of several different compound classes with known interference in different assay formats (e.g. Aggregators, Chelators, Clean, DMSO, Fluorescent, Luciferase, Reactive and Redox compounds) was pre-screened to test for liabilities with regard to certain classes of nuisance compounds. This approach showed that fluorescent compounds cause false positives by carry-over of signal in neighbouring wells. Therefore, we have implemented specific deselection assay(s) in our hit triaging cascade. The assay performed well yielding Z’>0.7 and a signal-to-noise ratio > 6.0.
In the HTS campaign the full deck of 450,000 compounds was screened to identify selective agonists for the ion channel. After active confirmation and de-selection the potency of the compounds were tested in Dose Response Curves (DRC).
Steven van Helden– Chief Technology Officer, Pivot Park Screening Centre B.V., Oss, Noord-Brabant, Netherlands
Chief Technology Officer
Pivot Park Screening Centre B.V.
Oss, Noord-Brabant, Netherlands
Steven van Helden studied chemistry at Utrecht University and, after obtaining his Ph.D. , worked in various roles in pharmaceutical industry for 20 years. Since 2003 he has been responsible for High Throughput Screening (HTS) operations and strategy at Organon/MSD. After the closure of those research facilities he developed a business plan for continuation of the screening activities in Oss, The Netherlands. This led to the formation of the Pivot Park Screening Centre (PPSC) and a central role of this company in the European Lead Factory. Steven is now Chief Technology Officer at PPSC.