Bead-Based Extracellular Vesicle Analysis Using Flow Cytometry.

Bead-Based Extracellular Vesicle Analysis Using Flow Cytometry.
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DOI:
10.1002/adbi.202000203
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发表时间:
2020-12
影响因子:
4.1
通讯作者:
Castro CM
Castro CM
中科院分区:
生物学3区
文献类型:
--
作者:
Yang KS;Lin HY;Curley C;Welch MW;Wolpin BM;Lee H;Weissleder R;Im H;Castro CM

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细胞外小泡(EV)是很有希望的癌症循环生物标志物,但由于缺乏简单、快速和可靠的EV分析,它们在临床环境中的高通量分析被证明是具有挑战性的。在这里,我们描述了一种基于微珠的EV流式细胞仪检测方法,它将使用微珠捕获EV与通过流式细胞仪进行EV蛋白分析相结合。该分析快速(48个样本4小时),稳健,并与用于高通量EV分析的传统流式细胞仪兼容。利用该方法,我们成功地分析了胰腺癌患者血浆样本中EVS中的一组胰腺癌生物标志物。该分析可以很容易地翻译成其他生物标志物或癌症类型,并可以在传统的流式细胞仪上与标准材料一起运行,使其具有高度的灵活性和适应性,适合不同的研究和临床需求。通过流式细胞术检测的基于微珠的EV检测能够快速、可靠、与传统的用于高通量EV分析的流式细胞仪兼容。该方法结合了用聚苯乙烯(PS)小球捕获EV和用流式细胞仪分析EV蛋白的方法。该分析可以在常规流式细胞仪上使用标准材料运行,使其具有高度的灵活性和适应性,适合不同的研究和临床需求。
Extracellular vesicles (EVs) represent promising circulating biomarkers for cancers, but their high-throughput analyses in clinical settings prove challenging due to lack of simple, fast, and robust EV assays. Here, we describe a bead-based EV assay detected by flow cytometry, which integrates EV capture using microbeads with EV protein analyses by flow cytometry. The assay is fast (< 4 hrs for 48 samples), robust and compatible with conventional flow cytometry instruments for high-throughput EV analysis. With the method, we successfully analyzed a panel of pancreatic cancer biomarkers in EVs from plasma samples of pancreatic cancer patients. The assay is readily translatable to other biomarkers or cancer types and can be run with standard materials on conventional flow cytometers, making it highly flexible and adaptable to diverse research and clinical needs. A bead-based EV assay detected by flow cytometry enables fast, robust, compatible with conventional flow cytometry instruments for high-throughput EV analysis. The method combines EV capture using polystyrene (PS) beads with EV protein analyses by flow cytometry. The assay can be run with standard materials on conventional flow cytometers, making it highly flexible and adaptable to diverse research and clinical needs.
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