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Methods. 2015 Oct 1;87:75-82. doi: 10.1016/j.ymeth.2015.03.018. Epub 2015 Mar 31.

Proteomics characterization of exosome cargo.

Author information

1
Department of Biochemistry, Vanderbilt University School of Medicine, United States. Electronic address: k.schey@vanderbilt.edu.
2
Department of Medicine, Vanderbilt University School of Medicine, United States; Department of Pharmacology, Vanderbilt University School of Medicine, United States.
3
Department of Biochemistry, Vanderbilt University School of Medicine, United States.

Abstract

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.

KEYWORDS:

Mass spectrometry; Post-translational modifications; Proteomics

PMID:
25837312
PMCID:
PMC4591097
DOI:
10.1016/j.ymeth.2015.03.018
[Indexed for MEDLINE]
Free PMC Article

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