Proteomics characterization of exosome cargo

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

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Review

MeSH terms

  • Amino Acid Sequence
  • Annexins / genetics
  • Annexins / isolation & purification
  • Annexins / metabolism
  • Antigens, CD / genetics
  • Antigens, CD / isolation & purification
  • Antigens, CD / metabolism
  • Bibliometrics
  • Biological Transport
  • Biomarkers / metabolism
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / isolation & purification
  • DNA-Binding Proteins / metabolism
  • Endosomal Sorting Complexes Required for Transport / genetics
  • Endosomal Sorting Complexes Required for Transport / isolation & purification
  • Endosomal Sorting Complexes Required for Transport / metabolism
  • Exosomes / chemistry*
  • Humans
  • Mass Spectrometry / methods*
  • Molecular Sequence Data
  • Protein Processing, Post-Translational*
  • Proteome / genetics
  • Proteome / isolation & purification*
  • Proteome / metabolism
  • Proteomics / instrumentation
  • Proteomics / methods*
  • Transcription Factors / genetics
  • Transcription Factors / isolation & purification
  • Transcription Factors / metabolism

Substances

  • Annexins
  • Antigens, CD
  • Biomarkers
  • DNA-Binding Proteins
  • Endosomal Sorting Complexes Required for Transport
  • Proteome
  • Transcription Factors
  • Tsg101 protein