Proteomics Unit, Spanish National Cancer Research Centre (CNIO), Madrid, Spain.
Cell Signaling and Clinical Proteomics Group. Biocruces Bizkaia Health Research Institute, Barkaldo, Spain.
Expert Rev Proteomics. 2022 Feb;19(2):89-101. doi: 10.1080/14789450.2022.2052849. Epub 2022 Mar 22.
The field of extracellular vesicles (EVs) is rapidly advancing. This progress is fueled by the applications of these agents as biomarkers and also as an attractive source to encapsulate therapeutics.
Different types of EVs, including exosomes, and other nanoparticles have been identified with key regulatory functions in cell-cell communication. However, the techniques used for their purification possess inherent limitations, resulting in heterogeneous preparations contaminated by other EVs subtypes and nano-size structures. It is therefore urgent to deconvolute the molecular constituents present in each type of EVs in order to accurately ascribe their specific functions. In this context, proteomics can profile, not only the lumen proteins and surface markers, but also their post-translational modifications, which will inform on the mechanisms of cargo selection and sorting.
Mass spectrometry-based proteomics is now a mature technique and has started to deliver new insights in the EV field. Here, we review recent developments in sample preparation, mass spectrometry (MS) and computational analysis and discuss how these advances, in conjunction with improved purification protocols, could impact thecharacterization of the complex landscape of EVs and other secreted nanoparticles.
细胞外囊泡（EVs）领域正在迅速发展。这一进展的推动因素是这些制剂作为生物标志物的应用，以及作为封装治疗剂的有吸引力的来源。
不同类型的 EVs，包括外泌体和其他纳米颗粒，已被确定在细胞间通讯中具有关键的调节功能。然而，用于其纯化的技术存在固有局限性，导致异质制剂被其他 EVs 亚型和纳米大小的结构污染。因此，迫切需要解析每种 EVs 中存在的分子成分，以便准确地将其特定功能归因于它们。在这种情况下，蛋白质组学不仅可以对腔蛋白和表面标志物进行分析，还可以对其翻译后修饰进行分析，从而为货物选择和分类的机制提供信息。
基于质谱的蛋白质组学现在是一项成熟的技术，并已开始在 EV 领域提供新的见解。在这里，我们回顾了样品制备、质谱（MS）和计算分析的最新进展，并讨论了这些进展如何与改进的纯化方案相结合，可能会影响 EV 和其他分泌的纳米颗粒的复杂景观的特征。
