Department of Medical Oncology, OncoProteomics Laboratory, Cancer Center Amsterdam, Amsterdam UMC, Location VUMC, Amsterdam, The Netherlands.
Department of Urology, Amsterdam UMC, Location VUMC, Amsterdam, The Netherlands.
Methods Mol Biol. 2023;2718:235-251. doi: 10.1007/978-1-0716-3457-8_13.
Urinary extracellular vesicles (uEVs) are a rich source of noninvasive protein biomarkers. However, for translation to clinical applications, an easy-to-use uEV isolation protocol is needed that is compatible with proteomics. Here, we provide a detailed description of a quick and clinical applicable uEV isolation protocol. We focus on the isolation procedure and subsequent in-depth proteome characterization using LC-MS/MS-based proteomics. As an example, we show how differential analyses can be performed using urine samples obtained from prostate cancer patients, compared to urine from controls.
尿细胞外囊泡 (uEVs) 是一种丰富的非侵入性蛋白质生物标志物来源。然而，为了将其转化为临床应用，需要一种易于使用的 uEV 分离方案，该方案与蛋白质组学兼容。在这里，我们提供了一种快速且适用于临床的 uEV 分离方案的详细描述。我们专注于分离过程以及使用基于 LC-MS/MS 的蛋白质组学进行的后续深入蛋白质组学特征分析。作为一个例子，我们展示了如何使用从前列腺癌患者获得的尿液样本与对照尿液进行差异分析。
