期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 413, 期 2, 页码 365-375出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-020-03004-w
关键词
Saliva; Extracellular vesicles; Proteomics; Ultracentrifugation; Polymer-based precipitation
资金
- National Natural Science Foundation of China [31741036, 31700702, 21904098, U1703251, U1810113]
- Key Research and Development Plan of Shaanxi Province [2017ZDCXL-GY-10-0102]
- Key Laboratory of Coal Resources Exploration and Comprehensive Utilization, Ministry of Land and Resources Open Research Topic [KF2016-4]
- Shaanxi Provincial Innovation Capability Support Program [2019TD-021]
- Zhejiang Provincial and Ministry of Health Research Fund for Medical Sciences [WKJ-ZJ-1910]
- Wenzhou Medical University [89218012, 89219012]
Salivary extracellular vesicles obtained by ultracentrifugation and polyethylene glycol precipitation methods were compared in this study, with the former showing higher expression of EV-specific markers. Analysis using tandem mass spectrometry proteomics revealed that ultracentrifugation can isolate more EV-related proteins, providing guidance for EV separation and potential applications in non-invasive diagnosis.
Salivary extracellular vesicles (EVs), as novel functional carriers and potential biomarkers, are usually obtained by ultracentrifugation (UC) and polyethylene glycol (PEG)-based precipitation methods. However, salivary EVs obtained by these two methods have not been systematically compared. Here, we perform an in-depth analysis on EVs isolated by these two methods using proteomics. Both methods obtain EVs ranging from 40 to 210 nm, with the PEG method resulting in a wider size distribution. PEG-separated products were irregularly shaped and aggregated, while UC-separated ones were monodispersed and teacup-shaped. Additionally, the expression of EV-specific markers was higher in UC-separated EVs. Using tandem mass spectrometry proteomics, we identified and quantified 1217 kinds of saliva exosomal proteins and 361 kinds of differential proteins, showing that UC can isolate more EV-related proteins. These results offer some guidance for EV separating and provide potential direction for the use of EVs in non-invasive diagnosis.
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