Improving immunological stability of EVs through lipoprotein depletion in plasma 9128

Abstract Description Background Extracellular vesicles (EVs) regulate physiological and pathological processes, and plasma-derived EVs have significant potential as biomarkers for various diseases. However, lipoproteins in plasma act as major obstacles, compromising EV purity and immunological reliability. Lipoproteins induce inflammatory responses, mimic EVs physically, and trigger unexpected immune reactions, complicating downstream applications. Methods This study proposes a novel method using antibodies against Apolipoprotein A1 and Apolipoprotein B to selectively remove lipoproteins from plasma. Following lipoprotein removal, EVs were isolated using a precipitation kit. Results Nanoparticle tracking analysis (NTA) confirmed EVs with an average size of approximately 190 nm, and scanning electron microscopy (SEM) revealed the characteristic cup-shaped structure of EVs. Flow cytometry (FACS) analysis showed a significant decrease in Apo A1 signals in the lipoprotein-depleted group, while EV-positive markers (CD9, CD63) were maintained. Western blot analysis similarly demonstrated reduced Apo A1 bands after lipoprotein removal. Conclusion This antibody-based lipoprotein removal approach provides a promising strategy for obtaining highly pure EVs and minimizing immunological interference. It is expected to significantly enhance the utility of EV-based research and clinical diagnostics. Funding Sources Supported by KGM4572532 Topic Categories Technological Innovations in Immunology (TECH)