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The importance of extracellular vesicle purification for downstream analysis: A comparison of differential centrifugation and size exclusion chromatography for helminth pathogens

机译:细胞外囊泡纯化对下游分析的重要性:蠕虫病原体的差速离心和尺寸排阻色谱比较

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摘要

BackgroundRobust protocols for the isolation of extracellular vesicles (EVs) from the rest of their excretory-secretory products are necessary for downstream studies and application development. The most widely used purification method of EVs for helminth pathogens is currently differential centrifugation (DC). In contrast, size exclusion chromatography (SEC) has been included in the purification pipeline for EVs from other pathogens, highlighting there is not an agreed research community ‘gold standard’ for EV isolation. In this case study, Fasciola hepatica from natural populations were cultured in order to collect EVs from culture media and evaluate a SEC or DC approach to pathogen helminth EV purification.
机译:背景技术用于下游研究和应用开发所必需的用于将细胞外囊泡(EV)与它们的其余排泄-分泌产物分离的稳健方案是必需的。目前,用于蠕虫病原体的电动汽车最广泛的纯化方法是差速离心(DC)。相比之下,尺寸排阻色谱(SEC)已包含在其他病原体电动车的纯化管线中,这突显出目前尚无公认的电动车隔离研究“金标准”。在本案例研究中,培养了自然种群的Fasciola hepatica,以便从培养基中收集EV,并评估SEC或DC方法纯化病原体蠕虫EV。

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