Somchai ChutipongtanateKenneth D. GreisUniversity of Cincinnati College of MedicineFaculty of Medicine, Ramathibodi Hospital, Mahidol University2019-08-282019-08-282018-12-01Scientific Reports. Vol.8, No.1 (2018)204523222-s2.0-85054772012https://repository.li.mahidol.ac.th/handle/20.500.14594/47475© 2018, The Author(s). 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.Mahidol UniversityMultidisciplinaryArticleSCOPUS10.1038/s41598-018-33280-7