Microenvironment and Metastasis Laboratory, Molecular Oncology Programme, Spanish National Cancer Research Center (CNIO), Madrid, Spain.
Proteomics Unit-ProteoRed-ISCIII, Spanish National Cancer Research Centre (CNIO), Madrid, Spain.
Methods Mol Biol. 2021;2265:345-359. doi: 10.1007/978-1-0716-1205-7_25.
Circulating extracellular vesicles in biofluids have become an interesting approach to analyse disease biomarkers. There are multiple methods for isolation of extracellular vesicles, though differential ultracentrifugation is still considered as the gold-standard isolation technique for exosomes. Furthermore, exosomes purified by this method have been demonstrated to display functional activity in vitro and in vivo and exhibit great versatility for subsequent analysis including proteomics, electron microscopy, mass spectrometry, or nucleic acid analysis. Here, we describe the method for isolation of exosomes from lymphatic exudate (seroma) obtained postlymphadenectomy for liquid biopsy approaches.
在生物体液中循环的细胞外囊泡已成为分析疾病生物标志物的一种有趣方法。有多种方法可用于分离细胞外囊泡，尽管差速超速离心仍然被认为是外泌体的金标准分离技术。此外，通过该方法纯化的外泌体已被证明在体外和体内具有功能活性，并表现出极大的多功能性，可用于随后的分析，包括蛋白质组学、电子显微镜、质谱或核酸分析。在这里，我们描述了从淋巴液渗出物（血清肿）中分离外泌体的方法，该方法是在淋巴结切除术后获得的，用于液体活检。
