广西医学

目的对比三种白花蛇舌草来源外囊泡（HDW⁃EVLPs）提取方法的优劣性。方法使用超速离心法、全自动外泌体提纯系统EXODUS和3D FloTrix® vivaEXO外泌体收获系统提取HDW⁃EVLPs，对比三种方法单次提取的性能指标、经济性与适用性指标、HDW⁃EVLPs表征、 HDW⁃EVLPs平均颗粒蛋白比和平均颗粒 RNA 比等关键指标。结果 3D FloTrix® vivaEXO外泌体收获系统单次提取的最大上样量和总颗粒数最高，分别为10 000 mL、1.44×1012 Particles，远超超速离心法和全自动外泌体提纯系统EXODUS。同时，该方法的设备市场价最低但单次耗材成本最高。三种方法提取的HDW⁃EVLPs形态学特征相似，均呈现典型的双层膜“茶托样”结构。全自动外泌体提纯系统EXODUS提取的HDW⁃EVLPs的平均颗粒浓度、平均 RNA 含量与超速离心法相比，差异有统计学意义（P<0.05）；全自动外泌体提纯系统EXODUS提取的HDW⁃EVLPs的平均 RNA 含量、平均蛋白含量与3D FloTrix® vivaEXO外泌体收获系统相比，差异有统计学意义（P<0.05）。三种方法提取的HDW⁃EVLPs的平均颗粒蛋白比和平均颗粒RNA比的数量级相同。结论三种方法提取的HDW⁃EVLPs在基本特性和结构完整性方面表现相似，但相关关键指标存在差异，用户可根据自身的需求和预算，选择最合适的HDW⁃EVLPs提取方法。

Objective To compare the advantages and disadvantages of three extraction methods for Hedyotis diffusa willd⁃derived extracellular vesicle⁃like particles (HDW⁃EVLPs). Methods HDW⁃EVLPs were extracted using ultracentrifugation, the fully automatic exosome purification system EXODUS, and the 3D FloTrix® vivaEXO exosome harvesting system. Key performance indicators for a single extraction, economic and applicability metrics, HDW⁃EVLPs characterization, HDW⁃EVLPs average particle⁃to⁃protein ratio, and average particle⁃to⁃RNA ratio of HDW⁃EVLPs, were compared between the three methods. Results The 3D FloTrix® vivaEXO exosome harvesting system achieved the highest maximum loading volume and total particle number per extraction, at 10 000 mL and 1.44×1012 Particles, respectively, far exceeding those of ultracentrifugation and the fully automatic exosome purification system EXODUS. Meanwhile, this method had the lowest market price for equipment but the highest cost per run for consumables. The morphological characteristics of HDW⁃EVLPs extracted by all three methods were similar, exhibiting the typical bilayer membrane “saucer⁃like” structure. Compared to ultracentrifugation, the fully automatic exosome purification system EXODUS exhibited statistically significant differences in average particle concentration and average RNA content (P<0.05). Compared to the 3D FloTrix® vivaEXO exosome harvesting system, the fully automatic exosome purification system EXODUS indicated statistically significant differences in average RNA content and average protein content of HDW⁃EVLPs (P<0.05). The average particle⁃to⁃protein ratios and average particle⁃to⁃RNA ratios of HDW⁃EVLPs extracted by the three methods were of the same order of magnitude. Conclusion HDW⁃EVLPs extracted by the three methods exhibit similar basic characteristics and structural integrity, but differences exist in key performance indicators. Users can select the most suitable HDW⁃EVLPs extraction method based on their specific needs and budget.

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