摘要Elapid snakebites cause severe toxicity,predominantly neurotoxicity and general cytotoxicity.However,the specific cellular impacts of individual venom toxins remain largely underexplored.This study developed a high-throughput platform for profiling cytotoxicity from elapid venoms,focusing on nanofractionation analytics to enhance selectivity and toxin identification.Elapid Venoms were tested on four human cell lines,representing kidney(RPTEC/TERT1),liver(HepaRG),endothelial(iPSC-EC),and skin(HaCaT)tissues.Cytotoxic effects were assessed through cell coverage,viability,and metabolic assays in both crude and nanofractionated venom samples.Nanofractionation revealed selective cyto-toxicity in venom components,notably phospholipases A2(PLA2s)and three-finger toxins(3FTxs),which impaired membrane integrity and cellular metabolism.Crude B.multicinctus venom displayed specific cytotoxicity toward liver and skin cells but not kidney or endothelial cells.Cytotoxicity of nanofractionated B.multicinctus venom was lost,likely due to denaturing conditions of the reversed-phase separation.Fractionation after size exclusion chromatography(SEC)for post-column bio-assaying to avoid toxin denaturation yielded bioactive fractions,with 3FTxs,PLA2s,and Kunitz-type serine protease(KUNs)likely responsible for the observed cell permeability disruption,extracellular matrix(ECM)degradation,and metabolic loss.This integrated analytical workflow,combining nano-fractionation with high-throughput cytotoxicity assays and venomics,enabled rapid identification of venom components with cell type-specific toxicity.Our findings contribute to understanding elapid venom toxicity and can aid in developing targeted snakebite treatments focusing on cytotoxicity responsible for tissue-specific damage.