Exosomes are membrane‐secreted vesicles, with sizes ranging from 30 to 150 nm, which play key roles in intercellular communication. There is intense interest in developing methods to isolate and quantify exosomes toward clinical diagnostics, fundamental studies of intercellular processes, and use of exosomes as delivery vehicles for therapeutic agents. Current methods for exosomes isolation and quantification are time consuming and have operational high costs; few combine isolation and quantification into a singular operation unit. This report describes the use of hydrophobic interaction chromatography on a polyester capillary‐channeled polymer fiber column, employing a step gradient for exosome elution, including use of glycerol as a solvent modifier. The entire procedure is completed in 8 min, while maintaining the structural integrity and biological activity of the isolated exosomes. Electron microscopy was used to verify the size and structural fidelity of single exosomes. Absorbance response curves for a commercial exosome sample were used for exosome quantification in the chromatographic separations. In order to determine the dynamic loading capacity for exosomes, different volumes of Dictyostelium discoideum cell culture milieu supernatant were loaded at different column lengths (5–30 cm) and loading flow rates (0.2–0.5 ml/min). A loading capacity of 5.4 × 10¹² exosomes derived from D. discoideum milieu was obtained on a 0.8 × 300 mm column; yielding recoveries of over 80%. It is believed that this isolation and purification strategy holds many advantages toward the use of exosomes across a wide breadth of medical and biotechnology applications.

中文翻译：

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通过在聚酯、毛细管通道聚合物纤维相上通过甘油辅助疏水相互作用色谱法在纯化过程中评估外泌体负载特性。

外泌体是膜分泌的囊泡，大小从 30 到 150 nm，在细胞间通讯中起关键作用。人们对开发用于临床诊断、细胞间过程的基础研究以及使用外泌体作为治疗剂的递送载体的外泌体的分离和量化方法有着浓厚的兴趣。目前的外泌体分离和定量方法耗时且操作成本高；很少有人将隔离和量化结合到一个单一的操作单元中。本报告描述了在聚酯毛细管聚合物纤维柱上使用疏水相互作用色谱，采用阶梯梯度进行外泌体洗脱，包括使用甘油作为溶剂改性剂。整个过程在8分钟内完成，同时保持分离的外泌体的结构完整性和生物活性。电子显微镜用于验证单个外泌体的大小和结构保真度。商业外泌体样品的吸光度响应曲线用于色谱分离中的外泌体定量。为了确定外泌体的动态负载能力，不同体积的盘基网柄菌细胞培养环境上清液以不同的柱长（5-30 厘米）和加载流速（0.2-0.5 毫升/分钟）加载。在 0.8 × 300 mm 柱上获得了来自D. discoideum环境的 5.4 × 10 ¹²外泌体的负载能力。收率超过80%。相信这种分离和纯化策略对于在广泛的医学和生物技术应用中使用外泌体具有许多优势。