Abstract

Extracellular vesicles (EVs) and cell-derived vesicles (CDVs), generated by fragmenting cellular membranes, have both been explored as therapeutic delivery vehicles. Surface proteins on these vesicles are of great importance as they are characteristic to the cell of origin and modulate vesicle interactions with target cells. Here, we introduced a high-throughput fluorescence correlation spectroscopy (ht-FCS) approach capable of characterizing vesicle surface proteins across a large number of samples. We used automated screening and acquisition of FCS data to profile surface proteins of cell-derived vesicles with high fidelity based on changes in diffusion time upon antibody-vesicle interactions. We characterized vesicles generated from 4 cell types using antibodies for known exosome biomarkers. The ht-FCS technique presented here offers the capability to screen EVs or cell-derived vesicles against a library of surface markers or to screen a library of cell-derived vesicles for a specific identifying marker at a high speed.

中文翻译：

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高通量荧光相关光谱能够分析细胞衍生囊泡的表面成分

摘要

细胞外囊泡 (EV) 和细胞衍生囊泡 (CDV) 是由细胞膜破碎产生的，都已被探索作为治疗性递送载体。这些囊泡上的表面蛋白非常重要，因为它们是起源细胞的特征并调节囊泡与靶细胞的相互作用。在这里，我们介绍了一种高通量荧光相关光谱 (ht-FCS) 方法，该方法能够表征大量样品中的囊泡表面蛋白。我们使用自动筛选和获取 FCS 数据，根据抗体-囊泡相互作用时扩散时间的变化，以高保真度分析细胞衍生囊泡的表面蛋白。我们使用已知外泌体生物标志物的抗体对从 4 种细胞类型产生的囊泡进行了表征。