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Although seminal exosomes have been identified in human seminal plasma, their exact composition and possible physiologic function remain unknown. The objective of this study was to perform a comprehensive proteomics analysis of exosomes derived from human seminal plasma. Seminal exosomes were isolated and purified from 12 healthy donors using a 30% sucrose cushion\u2010based exosome\u2010isolation protocol, followed by characterization by western blot, transmission electron microscopy, and nanoparticle tracking analysis before performing extensive liquid chromatography tandem mass spectrometry proteomics analysis. The identified proteins were analyzed by bioinformatics analysis, and seminal exosomes\u2010associated proteins were selectively validated by western blot. A total of 1474 proteins were identified in all seminal exosomes samples, with Gene Ontology analysis demonstrating that these identified seminal exosomes\u2010associated proteins were mostly linked to \u2018exosomes,\u2019 \u2018cytoplasm,\u2019 and \u2018cytosol.\u2019 Bioinformatics analysis indicated that these proteins were mainly involved in biologic processes, including metabolism, energy pathways, protein metabolism, cell growth and maintenance, and transport. Of these identified proteins, PHGDH, LGALS3BP, SEMG1, ACTB, GAPDH, and the exosomal\u2010marker protein ALIX were validated by western blot. This study provided a more comprehensive description of the seminal exosomes proteome and could also be a resource for further screening of biomarkers and comparative proteomics studies, including those associated with male infertility and prostate cancer.<\/jats:p>","DOI":"10.1111\/andr.12412","type":"journal-article","created":{"date-parts":[[2017,9,15]],"date-time":"2017-09-15T10:10:25Z","timestamp":1505470225000},"page":"1007-1015","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":124,"title":["Comprehensive proteomics analysis of exosomes derived from human seminal plasma"],"prefix":"10.1111","volume":"5","author":[{"given":"C.","family":"Yang","sequence":"first","affiliation":[{"name":"Department of Urology The Third Affiliated Hospital of Southern Medical University Guangzhou 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