Comparison of different methods for isolating CD8 ⁺ T lymphocyte‐derived extracellular vesicles and supramolecular attack particles

CD8+ T lymphocytes play vital roles in killing infected or deranged host cells, recruiting innate immune and regulating other aspects of responses. Like any cell, cells also produce extracellular particles. These include vesicles (EVs) non-vesicular particles (NVEPs). cell-derived EVs are proposed to mediate cell-to-cell signalling, especially the context inflammatory responses, autoimmunity, infectious diseases. supramolecular attack (SMAPs), which same size range as a component cell mediated killing. The isolation technique selected will have profound effect on yield, purity, biochemical properties function particles; making it important directly compare different approaches. In this study, we compared commonly used techniques (membrane spin filtration, ultracentrifugation, exclusion liquid chromatography) isolate from activated human validated our results by single-particle methods, including nanoparticle tracking analysis, flow cytometry, electron microscopy super-resolution purified sample well bulk proteomics lipidomics analyses evaluate quality nature enriched Our show that there is trade-off between yield Furthermore, protein lipid composition dramatically impacted applied. We conclude evaluated, chromatography offers highest derived SMAPs with acceptable yields for compositional functional studies.