Assay Development and Screening
Philip Kelly, BSc
Screening Multiple PPI Targets in Parallel: Accelerating Portfolio Decisions
Charles River
SAFFRON WALDEN, England, United Kingdom
Screening Multiple PPI Targets in Parallel: Accelerating Portfolio-level Decisions
Phil Kelly, Senior Scientist & Gurvan Mahe, Scientist
Abstract: In a fast-past evolving world, high quality data and trustworthy partnerships are the key component to success. At Charles River, we developed within few months’ multiple high throughput screening projects in collaboration with our client to detect small molecule inhibitors of protein-protein interaction (PPI) targets. The assays were performed in parallel in a unique cross-site collaboration between the UK and the Netherlands.
The hit-finding campaign initially started with 3 targets (receptor/cytokine). All assays were developed in 1536-well plate format with a TR-FRET readout, where possible assays utilized the same detection reagents allowing targets to be used as counter-screened against each other. Three libraries were screened on each target for a total of 370k compounds, followed by hit confirmation potency and SPR studies.
After hit selection and outlier’s exclusion, a combined effort from medicinal chemists from Charles River and LEO Pharma allowed the annotation, clustering, and prioritization of hits.
During the screening campaign some unforeseen plate effects were observed which varied between targets. Troubleshooting was performed and the effects were directly linked to dry ice shipments and plate types used. Adjustments were made to our processes and findings were shared with the wider group to prevent any further issues when screening
To conclude, this partnership allowed the identification of multiple auspicious hits on different targets. The original project has expanded to additional receptors, and the hunt for inhibitors continues.
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