ポスター：Eurofins Discovery社、ヒット化合物探索の革新的アプローチ - ハイスループットスペクトルシフトを用いた結合ベースの低分子スクリーニング
SLAS, San Diego, CA, USA
1月 27, 2025
結合ベースのスクリーニングにおける専門性を持つ同社では、膨大かつ多様な低分子ライブラリーから迅速にヒット化合物をスクリーニングするため、NanoTemper社Dianthus uHTS装置およびGenedata Screener®を用いて、ハイスループットスペクトルシフトによる高速データ処理システムを開発しました。
For years, Eurofins Discovery has been investing in Hit Finding technologies for small molecules and fragment discovery. Our team of experts strongly believes in the added value of biophysical methods and direct target-binding assessment in the early stages of the drug discovery process. However, up to now, biophysics were instrumental at the hit validation stage after compound screening based on potency and selectivity. The biophysical methods have not been amenable to large scale primary screening due to their limitations in sample requirements, throughput or robotic integration. The cutting-edge Dianthus uHTS equipment created by NanoTemper Technologies, coupled with Eurofins Discovery’s expertise in affinity screening brings major innovation to the hit finding paradigm. For the first time, binders can be identified in large-scale single dose screening using a direct biophysical measurement. The NanoTemper Dianthus uHTS is a 1536-well instrument that relies on Spectral Shift (SpS) technology, an immobilization-free detection method that uses precision optics to monitor changes in fluorescence intensity during biomolecular interactions under isothermal conditions. In addition to the 1536-well miniaturization, the Dianthus uHTS can be fully integrated with a robotic platform. The combination of these technologies makes it possible to perform primary screening with the exceptional throughput of more than 25,000 compounds per day. Eurofins Discovery’s scientists first validated the Dianthus uHTS’ performance using a subset of hits that had previously been validated using multiple binding and functional assays. We analyzed protein stability, signal-to-noise, Z’ and affinity ranking values from compound titration, and concluded that assay sensitivity and reproducibility was similar to Dianthus 384-well instrument. We developed automated plate processing by combining the Dianthus uHTS with Genedata Screener®, in order to screen a diverse set of 50,000 small molecule library on the PIM3 kinase. In parallel, we generated screening data using the 1536-well SpS technology and the ADP-Glo™ enzymatic assay. The affinity-based screening strategy hit list nicely correlated with the PIM3 activity hits, but had the added benefit of lacking the common false-positives such as luciferase inhibitors. Additionally, early-stage MoA can be conducted, allowing for the possibility of identifying more characterized hits – leading to the initiation of successful hit-to-lead program. In recent years, Eurofins Discovery has employed hit finding technologies such as RapidFire-MS, Echo® MS and state-of-the art biophysical methods such as SPR, MST or nanoDSF. With its unprecedented throughput, the Dianthus uHTS platform offers a major breakthrough in early drug discovery, enabling the identification of direct binders at the primary screening stage. It provides a valuable alternative to ASMS and DEL, making high-throughput, site-agnostic binding approaches achievable with a very small amount of the tested protein, no data deconvolution, and with no need of compound multiplexing.
