biostudies-literatureWatanabe DMEXT | Japan Society for the Promotion of ScienceMEXT | Japan Society for the Promotion of Science (JSPS)Japan Agency for Medical Research and DevelopmentMinistry of Education, Culture, Sports, Science and TechnologyMEXT | JST | Core Research for Evolutional Science and TechnologyMEXT | JST | Core Research for Evolutional Science and Technology (CREST)Ministry of Education, Culture, Sports, Science and Technology (MEXT)Japan Agency for Medical Research and Development (AMED)8975https://www.ebi.ac.uk/biostudies/studies/S-EPMC11486946biostudies-literatureUnknown15(1)The single-molecule tracking of transmembrane receptors in living cells has provided significant insights into signaling mechanisms, such as mobility and clustering upon their activation/inactivation, making it a potential screening method for drug discovery. Here we show that single-molecule tracking-based screening can be used to explore compounds both detectable and undetectable by conventional methods for disease-related receptors. Using an automated system for a fast large-scale single-molecule analysis, we screen for epidermal growth factor receptor (EGFR) from 1134 of FDA approved drugs. The 18 hit compounds include all EGFR-targeted tyrosine kinase inhibitors (TKIs) in the library that suppress any phosphorylation-dependent mobility shift of EGFR, proving the concept of this approaNature communicationsSingle molecule tracking based drug screening.PMC1148694618H0541418H01839JP23ym0126815JPMJCR21E122H02593Ueda MWatanabe DYasui MHiroshima MfalseSingle molecule tracking based drug screening.The single-molecule tracking of transmembrane receptors in living cells has provided significant insights into signaling mechanisms, such as mobility and clustering upon their activation/inactivation, making it a potential screening method for drug discovery. Here we show that single-molecule tracking-based screening can be used to explore compounds both detectable and undetectable by conventional methods for disease-related receptors. Using an automated system for a fast large-scale single-molecule analysis, we screen for epidermal growth factor receptor (EGFR) from 1134 of FDA approved drugs. The 18 hit compounds include all EGFR-targeted tyrosine kinase inhibitors (TKIs) in the library that suppress any phosphorylation-dependent mobility shift of EGFR, proving the concept of this approa2024-01-01T00:00:00Z2024 Oct2025-04-04T20:55:24.622Z2025-04-04T20:55:24.622ZS-EPMC114869463942001510.1038/s41467-024-53432-w