Ko Jina, Wang Yongcheng, Sheng Kuanwei, Weitz David A, Weissleder Ralph
Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, CPZN 5206, Boston, Massachusetts 02114, United States.
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, United States.
ACS Nano. 2021 Mar 23;15(3):5631-5638. doi: 10.1021/acsnano.1c00782. Epub 2021 Mar 9.
Circulating extracellular vesicles (EVs)-biological nanomaterials shed from most mammalian cells-have emerged as promising biomarkers, drug delivery vesicles, and treatment modulators. While different types of vesicles are being explored for these applications, it is becoming clear that human EVs are quite heterogeneous even in homogeneous or monoclonal cell populations. Since it is the surface EV protein composition that will largely dictate their biological behavior, high-throughput single EV profiling methods are needed to better define EV subpopulations. Here, we present an antibody-based immunosequencing method that allows multiplexed measurement of protein molecules from individual nanometer-sized EVs. We use droplet microfluidics to compartmentalize and barcode individual EVs. The barcodes/antibody-DNA are then sequenced to determine protein composition. Using this highly sensitive technology, we detected specific proteins at the single EV level. We expect that this technology can be further adapted for multiplexed protein analysis of any nanoparticle.
循环细胞外囊泡（EVs）——从大多数哺乳动物细胞中脱落的生物纳米材料——已成为有前景的生物标志物、药物递送囊泡和治疗调节剂。虽然正在探索不同类型的囊泡用于这些应用，但越来越清楚的是，即使在同质或单克隆细胞群体中，人类EVs也相当异质。由于正是表面EV蛋白组成将在很大程度上决定其生物学行为，因此需要高通量单EV分析方法来更好地定义EV亚群。在这里，我们提出了一种基于抗体的免疫测序方法，该方法允许对单个纳米级EVs中的蛋白质分子进行多重测量。我们使用液滴微流控技术对单个EVs进行分隔和条形码标记。然后对条形码/抗体-DNA进行测序以确定蛋白质组成。使用这种高度灵敏的技术，我们在单EV水平上检测到了特定蛋白质。我们期望该技术可以进一步适用于任何纳米颗粒的多重蛋白质分析。
