biostudies-literatureCheng HNational Cancer InstituteNCI NIH HHS113368https://www.ebi.ac.uk/biostudies/studies/S-EPMC8321683biostudies-literatureUnknown190D-2-hydroxyglutarate (D2HG) is over-produced as an oncometabolite due to mutations in isocitrate dehydrogenases (IDHs). Accumulation of D2HG can cause the dysfunction of many enzymes and genome-wide epigenetic alterations, which can promote oncogenesis. Quantification of D2HG at single-cell resolution can help understand the phenotypic signatures of IDH-mutant cancers and identify effective therapeutics. In this study, we developed an analytical method to detect D2HG levels in single cancer cells by adapting cascade enzymatic reactions on a resazurin-based fluorescence reporter. The resazurin probe was immobilized to the sensing surface via biotin-streptavidin interaction. This surface chemistry was rationally optimized to translate the D2HG levels to sensitive fluorescence readouts efficiBiosensors & bioelectronicsSingle-cell profiling of D-2-hydroxyglutarate using surface-immobilized resazurin analogs.PMC8321683U01 CA217655R03 CA227352U54 CA199090Li ZGuo ZMo LWei WXue MCheng HShao SfalseSingle-cell profiling of D-2-hydroxyglutarate using surface-immobilized resazurin analogs.D-2-hydroxyglutarate (D2HG) is over-produced as an oncometabolite due to mutations in isocitrate dehydrogenases (IDHs). Accumulation of D2HG can cause the dysfunction of many enzymes and genome-wide epigenetic alterations, which can promote oncogenesis. Quantification of D2HG at single-cell resolution can help understand the phenotypic signatures of IDH-mutant cancers and identify effective therapeutics. In this study, we developed an analytical method to detect D2HG levels in single cancer cells by adapting cascade enzymatic reactions on a resazurin-based fluorescence reporter. The resazurin probe was immobilized to the sensing surface via biotin-streptavidin interaction. This surface chemistry was rationally optimized to translate the D2HG levels to sensitive fluorescence readouts effici2021-01-01T00:00:00Z2021 Oct2025-04-05T20:11:54.636Z2025-04-05T20:11:54.636ZS-EPMC83216833409836110.1016/j.bios.2021.113368