4.7 Article

Proteomics characterization of exosome cargo

Journal

METHODS
Volume 87, Issue -, Pages 75-82

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2015.03.018

Keywords

Proteomics; Post-translational modifications; Mass spectrometry

Funding

  1. NIH [HL100016, DK081662, UL1RR024975]
  2. NCATS/NIH (Vanderbilt Institute for Clinical and Translational Research) [UL1TR000445]
  3. Vanderbilt Mass Spectrometry Research Center

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Characterization of exosomal cargo is of significant interest because this cargo can provide clues to exosome biogenesis, targeting, and cellular effects and may be a source of biomarkers for disease diagnosis, prognosis and response to treatment. With recent improvements in proteomics technologies, both qualitative and quantitative characterization of exosomal proteins is possible. Here we provide a brief review of exosome proteomics studies and provide detailed protocols for global qualitative, global quantitative, and targeted quantitative analysis of exosomal proteins. In addition, we provide an example application of a standard global quantitative analysis followed by validation via a targeted quantitative analysis of urine exosome samples from human patients. Advantages and limitations of each method are discussed as well as future directions for exosome proteomics analysis. (C) 2015 Elsevier Inc. All rights reserved.

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