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      An Update on Isolation Methods for Proteomic Studies of Extracellular Vesicles in Biofluids

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          Abstract

          Extracellular vesicles (EVs) are lipid bilayer enclosed particles which present in almost all types of biofluids and contain specific proteins, lipids, and RNA. Increasing evidence has demonstrated the tremendous clinical potential of EVs as diagnostic and therapeutic tools, especially in biofluids, since they can be detected without invasive surgery. With the advanced mass spectrometry (MS), it is possible to decipher the protein content of EVs under different physiological and pathological conditions. Therefore, MS-based EV proteomic studies have grown rapidly in the past decade for biomarker discovery. This review focuses on the studies that isolate EVs from different biofluids and contain MS-based proteomic analysis. Literature published in the past decade (2009.1–2019.7) were selected and summarized with emphasis on isolation methods of EVs and MS analysis strategies, with the aim to give an overview of MS-based EV proteomic studies and provide a reference for future research.

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          Most cited references 64

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          Large-scale proteomics and phosphoproteomics of urinary exosomes.

          Normal human urine contains large numbers of exosomes, which are 40- to 100-nm vesicles that originate as the internal vesicles in multivesicular bodies from every renal epithelial cell type facing the urinary space. Here, we used LC-MS/MS to profile the proteome of human urinary exosomes. Overall, the analysis identified 1132 proteins unambiguously, including 177 that are represented on the Online Mendelian Inheritance in Man database of disease-related genes, suggesting that exosome analysis is a potential approach to discover urinary biomarkers. We extended the proteomic analysis to phosphoproteomic profiling using neutral loss scanning, and this yielded multiple novel phosphorylation sites, including serine-811 in the thiazide-sensitive Na-Cl co-transporter, NCC. To demonstrate the potential use of exosome analysis to identify a genetic renal disease, we carried out immunoblotting of exosomes from urine samples of patients with a clinical diagnosis of Bartter syndrome type I, showing an absence of the sodium-potassium-chloride co-transporter 2, NKCC2. The proteomic data are publicly accessible at http://dir.nhlbi.nih.gov/papers/lkem/exosome/.
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            Extracellular vesicles in physiological and pathological conditions.

            Body fluids contain surprising numbers of cell-derived vesicles which are now thought to contribute to both physiology and pathology. Tools to improve the detection of vesicles are being developed and clinical applications using vesicles for diagnosis, prognosis, and therapy are under investigation. The increased understanding why cells release vesicles, how vesicles play a role in intercellular communication, and how vesicles may concurrently contribute to cellular homeostasis and host defense, reveals a very complex and sophisticated contribution of vesicles to health and disease. Copyright © 2012 Elsevier Ltd. All rights reserved.
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              Extracellular Vesicles Provide a Means for Tissue Crosstalk during Exercise

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                Author and article information

                Contributors
                Role: Academic Editor
                Role: Academic Editor
                Role: Academic Editor
                Role: Academic Editor
                Role: Academic Editor
                Role: Academic Editor
                Journal
                Molecules
                Molecules
                molecules
                Molecules
                MDPI
                1420-3049
                27 September 2019
                October 2019
                : 24
                : 19
                Affiliations
                [1 ]School of Chemistry, Chemical Engineering and Life Science, Wuhan University of Technology, 122 luoshilu, Wuhan 430070, China; lij@ 123456whut.edu.cn (J.L.); 714769257@ 123456whut.edu.cn (X.H.)
                [2 ]School of Biological Science & Medical Engineering, Southeast University, No.2 Sipailou, Nanjing 210096, China; 213150742@ 123456seu.edu.cn
                Author notes
                [* ]Correspondence: yangchenxi@ 123456seu.edu.cn
                Article
                molecules-24-03516
                10.3390/molecules24193516
                6803898
                31569778
                © 2019 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

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