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Author Spotlight: Advancing the Analysis of Plasma Extracellular Vesicle Proteome for Cardiovascular Biomarker Studies
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Quantitative Proteomic Analysis of Biogenesis-Based Classification for Extracellular Vesicles.

Linwen Zhang1,2, Jeremie Parot3,4, Vincent A Hackley3

  • 1Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.

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|November 11, 2020
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Summary
This summary is machine-generated.

Extracellular vesicles (EVs) are complex and not strictly defined by size. This study uses protein markers to show that both microvesicles and exosomes can vary in size, challenging traditional classifications.

Keywords:
AF4QconCATsclassificationexosomesextracellular vesiclesmicrovesiclesmultiple reaction monitoringtargeted proteomics

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Nanotechnology

Background:

  • Extracellular vesicles (EVs) are traditionally classified as microvesicles (large, plasma membrane-derived) or exosomes (small, endosomal-derived).
  • This binary classification is insufficient to describe the complex composition of EV preparations.
  • Understanding EV biogenesis is crucial for interpreting their biological functions.

Purpose of the Study:

  • To investigate the complexity of EV origins and composition.
  • To utilize specific protein markers to differentiate between plasma membrane and endosomal membrane origins of EVs.
  • To challenge the strict size-based definitions of microvesicles and exosomes.

Main Methods:

  • Quantitative mass spectrometry was employed to simultaneously measure plasma membrane-specific and endosomal membrane-specific proteins in EV preparations.
  • Differential ultracentrifugation was used to isolate microvesicles and exosomes.
  • Sonicated platelets and multidetector asymmetrical-flow field-flow fractionation were used to model mixed EVs and verify purity of isolated fractions.

Main Results:

  • Quantitative mass spectrometry successfully measured membrane-specific proteins in both microvesicle and exosome fractions.
  • Analysis of EV preparations from human plasma and ARPE-19 cell medium revealed the presence of both marker types within fractions.
  • The study demonstrated that EVs do not adhere to strict size limitations, with both microvesicles and exosomes exhibiting a range of sizes.

Conclusions:

  • The traditional size-based classification of EVs into microvesicles and exosomes is an oversimplification.
  • EV preparations are heterogeneous, containing vesicles derived from both plasma and endosomal membranes, irrespective of their size.
  • Protein marker analysis provides a more accurate method for characterizing EV origins and composition.