One-Step SAR: Using High-Content Imaging to Drive Small Molecule Protein Degraders from Hit-to-Lead
High-content imaging (HCI) techniques have revolutionized the landscape of drug discovery, offering powerful tools to accelerate the identification and characterization of potential drug candidates. By combining the capabilities of automated microscopy, image analysis, and multiplexing, HCI enables the simultaneous quantification of numerous cellular features within a single experiment. This approach provides a wealth of information on cellular morphology, protein content, localization, and functional readouts, all of which are crucial for understanding disease mechanisms and evaluating drug responses. These techniques not only enhance the speed and efficiency of screening large compound libraries but also offer deeper insights into the mechanisms of action of potential therapeutics, ultimately accelerating the drug discovery process from target identification to lead optimization. Here, Curia describes a cost-effective high-content immunofluorescence workflow developed in an immortalized patient cell line for the identification of small molecule protein degraders using the Revvity Opera Phenix™ confocal imaging system. With maximized content and careful assay design, the resulting data provides a rich description of compound effects including on-target and off-pathway phenotypes along with cytotoxicity information – all in a single assay.