a Department of Genetics , Cell- and Immunobiology, Semmelweis University , Budapest , Hungary.
b MS Proteomics Research Group, Research Centre for Natural Sciences , Hungarian Academy of Sciences , Budapest Hungary.
Expert Rev Proteomics. 2017 Dec;14(12):1073-1090. doi: 10.1080/14789450.2017.1392244. Epub 2017 Oct 26.
Extracellular vesicles are emerging sources of biomarkers for modern preventive and precision medicine. Extracellular vesicles in body fluids offer a unique opportunity for integrative biomarker approaches due to their complex biocargo that includes proteins, lipids, nucleic acids and metabolites. Mass spectrometry-based proteomics data suggest that a significant portion of human proteins are sorted into extracellular vesicles and amenable for biomarker discovery schemes. Areas covered: this review focuses on key aspects of isolation, quality control and subsequent analysis of blood plasma- and conditioned medium-derived extracellular vesicle proteins, and summarizes the current state-of-the-art in the field. Furthermore, it provides introduction and guidelines for mass spectrometry-based proteomic analysis of extracellular vesicles. Expert commentary: Comparison of newly developed isolation and purification techniques with classical ultracentrifugation-based approaches are highly recommended. It is also essential to use multiple analytical approaches to characterize the isolated extracellular vesicles prior to characterization of their biocargo. Rigor in data reproducibility, critical data analysis, awareness of potential pitfalls, standardization and benchmarking are required for extracellular vesicle research to fulfil the current expectation that these subcellular structures can become a valid source of next generation biomarkers.
细胞外囊泡是现代预防医学和精准医学的新兴生物标志物来源。体液中的细胞外囊泡为综合生物标志物方法提供了独特的机会，因为它们的复杂生物货物包括蛋白质、脂质、核酸和代谢物。基于质谱的蛋白质组学数据表明，很大一部分人类蛋白质被分拣到细胞外囊泡中，适合用于生物标志物发现方案。
本综述重点介绍了从血浆和条件培养基中分离和后续分析细胞外囊泡蛋白的关键方面，并总结了该领域的最新进展。此外，它还为基于质谱的细胞外囊泡蛋白质组学分析提供了介绍和指南。
强烈建议将新开发的分离和纯化技术与基于经典超速离心的方法进行比较。在对其生物货物进行表征之前，必须使用多种分析方法来对分离的细胞外囊泡进行表征。为了满足当前的期望，即这些亚细胞结构可以成为下一代生物标志物的有效来源，细胞外囊泡研究需要严格的数据可重复性、关键数据分析、对潜在陷阱的认识、标准化和基准测试。
