Sun Yuchen, Saito Kosuke, Saito Yoshiro
Division of Medicinal Safety Science, National Institute of Health Sciences, Kawasaki City, Kanagawa, Japan.
Methods Mol Biol. 2022;2504:157-173. doi: 10.1007/978-1-0716-2341-1_12.
Lipidomics is an omics approach to comprehensively study lipid profiles in biological samples, such as plasma, serum, urine, and tissue specimens. Moreover, lipidomic analyses are useful for identifying novel lipid biomarkers, especially for various metabolic and malignant diseases in humans. Extracellular vesicles (EVs) are lipid bilayer-encapsulated nanoparticles secreted from various cells into the extracellular space. In particular, circulating EVs in the blood stream have attracted considerable research interest as they are considered the fingerprint of the cells from which they are secreted and are a promising source for less-invasive biomarker screening. Here, we describe the entire workflow for the lipidomic analysis of circulating EVs, including the methods for their purification from human plasma and serum, liquid chromatography coupled with high-resolution mass spectrometry-based lipid measurement, and data analyses for profiling EV lipids. Using this methodological workflow, over 260 lipid molecules belonging to the glycerophospholipid and sphingolipid groups can be detected.
脂质组学是一种组学方法，用于全面研究生物样品（如血浆、血清、尿液和组织标本）中的脂质谱。此外，脂质组学分析有助于识别新型脂质生物标志物，特别是针对人类的各种代谢和恶性疾病。细胞外囊泡（EVs）是从各种细胞分泌到细胞外空间的脂质双层包裹的纳米颗粒。特别是，血流中的循环EVs引起了相当大的研究兴趣，因为它们被认为是分泌它们的细胞的指纹，并且是用于微创生物标志物筛选的有前途的来源。在这里，我们描述了循环EVs脂质组学分析的整个工作流程，包括从人血浆和血清中纯化它们的方法、液相色谱与基于高分辨率质谱的脂质测量，以及用于分析EV脂质的数据分析。使用这种方法工作流程，可以检测到超过260种属于甘油磷脂和鞘脂组的脂质分子。
