Szigyártó Imola Cs, Singh Priyanka, Sonallya Tasvilla, Románszki Loránd, Mihály Judith, Wiener Zoltán, Bebesi Tímea, Mészáros Gábor, Keresztes Zsófia, Thompson Michael, Varga Zoltán, Beke-Somfai Tamás
HUN-REN Research Centre for Natural Sciences, Institute of Materials and Environmental Chemistry, Biomolecular Self-assembly Research Group, Budapest, Hungary.
Hevesy György PhD School of Chemistry, Eötvös Loránd University, Budapest, Hungary.
Methods Mol Biol. 2025;2908:225-238. doi: 10.1007/978-1-0716-4434-8_15.
Extracellular vesicles (EVs) are phospholipid bilayer-enclosed nanoparticles secreted by most cells into the biofluids. They have an important role in intercellular communication by carrying bioactive components (e.g., lipids, proteins, nucleic acids, and other metabolites) and reflecting the biochemical and metabolic processes of their parent cell. Due to their widespread molecular cargo, they have potential diagnostic and therapeutic roles, so they can be considered as new generation biomarkers. With the drastic increase in the number of publications and the comparability of the obtained results, the development of standardized protocols has become necessary. Here we aim to demonstrate the applicability of two, non-conventional techniques, such as ATR-infrared spectroscopy (ATR-FTIR) and electromagnetic piezoelectric acoustic sensor (EMPAS) in the characterization of compositional changes of blood- and pancreatic tumor-derived vesicles. In addition, we provide guidelines in sample collection, isolation, and separation of EVs. IR spectroscopy, as a fast and label-free technique, gives biochemical insight into the sample composition, while EMPAS serves information on vesicle quantity and in membrane integrity changes during storage.
细胞外囊泡（EVs）是大多数细胞分泌到生物流体中的磷脂双层包裹的纳米颗粒。它们通过携带生物活性成分（如脂质、蛋白质、核酸和其他代谢物）在细胞间通讯中发挥重要作用，并反映其母细胞的生化和代谢过程。由于其广泛的分子载荷，它们具有潜在的诊断和治疗作用，因此可被视为新一代生物标志物。随着出版物数量的急剧增加以及所得结果的可比性，制定标准化方案变得必要。在这里，我们旨在证明两种非传统技术，即衰减全反射红外光谱（ATR-FTIR）和电磁压电声学传感器（EMPAS）在表征血液和胰腺肿瘤衍生囊泡的成分变化方面的适用性。此外，我们提供了关于EVs样本收集、分离和纯化的指南。红外光谱作为一种快速且无标记的技术，能深入了解样品的生化组成，而EMPAS则提供有关囊泡数量以及储存期间膜完整性变化的信息。
