Extracellular vesicles (EVs) present a promising modality for numerous biological and medical applications, including therapeutics. Developing facile methods to engineer EVs is essential to meeting the rapidly expanding demand for various functionalized EVs in these applications. Herein, we developed a technology that integrates enzymatic glycoengineering and microfluidics for effective EV functionalization. This method builds on a 3D nanostructured microfluidic device to streamline a multiple-step EV engineering process, which involves a step of enzymatic reaction to install azido-sialic acid residues to glycans on EVs using a sialyltransferase and an azide-tagged sialyl donor followed by the attachment of various functionalities, such as biotin and fluorescent labels, to the resulting azido-glycans on EVs through a biocompatible click reaction. Compared to traditional EV engineering methods, we show that our technology improves the efficiency of EV glycoengineering while simplifying and expediting the workflow. Furthermore, we demonstrated the applicability of this technology to EVs derived from the cell lines of different cancer types, including A549, PC3, and COLO-1 cells. Overall, this EV engineering technology could provide a potentially useful tool for broad applications.

中文翻译：

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使用纳米材料接口微流体对细胞外囊泡进行高效的酶促聚糖工程


细胞外囊泡 （EV） 为许多生物和医学应用（包括治疗学）提供了一种很有前途的方式。开发简单的方法来设计 EV 对于满足这些应用中对各种功能化 EV 的快速增长需求至关重要。在此，我们开发了一种集成酶促糖工程和微流体的技术，以实现有效的 EV 功能化。该方法基于 3D 纳米结构微流控装置构建，以简化多步骤 EV 工程过程，其中包括一个酶促反应步骤，使用唾液酸转移酶和叠氮化物标记的唾液酸供体将叠氮基唾液酸残基安装到 EV 上的聚糖上，然后通过生物相容性点击反应将各种官能团（如生物素和荧光标记）连接到生成的 EV 上的叠氮基聚糖上。与传统的 EV 工程方法相比，我们表明我们的技术提高了 EV 糖工程的效率，同时简化和加快了工作流程。此外，我们证明了该技术对来自不同癌症类型细胞系（包括 A549、PC3 和 COLO-1 细胞）的 EV 的适用性。总体而言，这种 EV 工程技术可以为广泛的应用提供潜在的有用工具。