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引用本文:   刘海生, 袁文理, 陈永钰, 陈晨, 田野, 颜晓梅. 尿液细胞外囊泡的单颗粒水平多参数定量表征. 分析化学, 2020, 48(10): 1305-1314. doi:  10.19756/j.issn.0253-3820.201443 [复制]

Citation:   LIU Hai-Sheng , YUAN Wen-Li , CHEN Yong-Yu , CHEN Chen , TIAN Ye , YAN Xiao-Mei . Multi-parameter and Quantitative Characterization of Urinary Extracellular Vesicles at Single-Particle Level. Chinese Journal of Analytical Chemistry, 2020, 48(10): 1305-1314. doi: 10.19756/j.issn.0253-3820.201443 [复制]

尿液细胞外囊泡的单颗粒水平多参数定量表征

通讯作者:  颜晓梅, xmyan@xmu.edu.cn

收稿日期: 2020-07-29

基金项目: 本文系国家自然科学基金项目(Nos.21934004,21627811)和国家基础科学人才培养基金项目(No.J1310024)资助

Multi-parameter and Quantitative Characterization of Urinary Extracellular Vesicles at Single-Particle Level

Corresponding author:  YAN Xiao-Mei , xmyan@xmu.edu.cn

Received Date:  2020-07-29

Fund Project:  This work was supported by the National Natural Science Foundation of China (Nos. 21934004, 21627811) and the National Fund for Fostering Talents of Basic Science (No. J1310024).

尿液来源细胞外囊泡(Urinary extracellular vesicles,uEVs)作为新一类液体活检标志物,在疾病诊断、治疗监控领域具有广阔的应用前景。然而,由于缺乏相应的表征技术,研究者对于纳米尺度uEVs的粒径分布、颗粒浓度以及生化表型等均缺乏了解。EVs具有高度个体差异性和多样性,因此迫切需要发展高通量、多参数的单颗粒检测技术。基于本研究组前期工作中结合瑞利散射和鞘流单分子荧光检测技术研制的纳米流式检测装置(Nano-flow cytometer,nFCM),本研究对超速离心法纯化得到的uEVs的纯度、浓度、粒径、DNA、RNA和蛋白标志物在单颗粒水平进行多参数定量表征,其中EVs的检测限低至40 nm。结果表明,uEVs的纯度高达92%,10位健康受试者uEVs的颗粒浓度主要分布在3.0×109~8.9×1010个/mL之间,粒径主要分布在40~120 nm之间。DNA和RNA主要位于较大粒径的uEVs的内部,且颗粒之间的丰度存在很大的异质性。此外,使用nFCM对蛋白标志物CD9、CD63、CD81和CD24的阳性比率进行了测定。纳米流式检测技术可为uEVs的生化性质研究和临床应用提供重要的技术支撑。

关键词:   尿液细胞外囊泡, 纳米流式检测技术, 纯度, 粒径分布, 核酸, 蛋白标志物
Key words:   Urinary extracellular vesicles, Nano-flow cytometry, Purity, Size distribution, Nucleic acid, Protein marker
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尿液细胞外囊泡的单颗粒水平多参数定量表征

刘海生, 袁文理, 陈永钰, 陈晨, 田野, 颜晓梅

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