UC Proteomics Laboratory, Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, Ohio, United States.
Pediatric Translational Research Unit, Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
Sci Rep. 2018 Oct 9;8(1):15039. doi: 10.1038/s41598-018-33280-7.
The recent advance in targeted label-free proteomics, SWATH-MS, can provide consistent protein detection and reproducible protein quantitation, which is a considerable advantage for biomarker study of urinary extracellular vesicles. We developed a SWATH-MS workflow with a curated spectral library of 1,145 targets. Application of the workflow across nine replicates of three sample types (exosome-like vesicles (ELVs), microvesicles (MVs) and urine proteins (UPs)) resulted in the quantitation of 888 proteins at FDR <1%. The median-coefficient of variation of the 888 proteins in the ELV sample was 7.7%, indicating excellent reproducibility. Data analysis showed common exosome markers, (i.e. CD9, CD63, ALIX, TSG101 and HSP70) were enriched in urinary ELVs as compared to MVs and UPs. The use of a multiplex biomarker screening assay focused on ELVs was investigated, and perspectives in future applications are discussed.
近年来，靶向无标记蛋白质组学 SWATH-MS 的发展，可以提供一致的蛋白质检测和可重复的蛋白质定量，这是尿液细胞外囊泡生物标志物研究的一个重要优势。我们开发了一种具有经过精心整理的 1145 个靶标谱库的 SWATH-MS 工作流程。该工作流程在三种样本类型（类囊泡（ELVs）、微泡（MVs）和尿液蛋白（UPs））的 9 个重复中应用，结果定量了 888 个 FDR<1%的蛋白。ELV 样本中 888 个蛋白的中位数变异系数为 7.7%，表明具有极好的重现性。数据分析显示，与 MV 和 UP 相比，尿液 ELVs 中富含常见的外泌体标记物（即 CD9、CD63、ALIX、TSG101 和 HSP70）。还研究了针对 ELVs 的多重生物标志物筛选检测方法的应用，并讨论了未来应用的前景。
