A size-exclusion-based approach for purifying extracellular vesicles from human plasma

Autor: Gregory N. Ruegsegger, Mark W. Pataky, Patrick M. Vanderboom, Carrie J. Heppelmann, Fabrice Lucien, Surendra Dasari, K. Sreekumaran Nair, Ian R. Lanza
Rok vydání: 2021
Předmět:
Zdroj: Cell Reports: Methods, Vol 1, Iss 3, Pp 100055-(2021)
Cell reports methods
ISSN: 2667-2375
Popis: SUMMARY Extracellular vesicles (EVs) are released into blood from multiple organs and carry molecular cargo that facilitates inter-organ communication and an integrated response to physiological and pathological stimuli. Interrogation of the protein cargo of EVs is currently limited by the absence of optimal and reproducible approaches for purifying plasma EVs that are suitable for downstream proteomic analyses. We describe a size-exclusion chromatography (SEC)-based method to purify EVs from platelet-poor plasma (PPP) for proteomics profiling via high-resolution mass spectrometry (SEC-MS). The SEC-MS method identifies more proteins with higher precision than several conventional EV isolation approaches. We apply the SEC-MS method to identify the unique proteomic signatures of EVs released from platelets, adipocytes, muscle cells, and hepatocytes, with the goal of identifying tissue-specific EV markers. Furthermore, we apply the SEC-MS approach to evaluate the effects of a single bout of exercise on EV proteomic cargo in human plasma.
Graphical abstract
In brief Vanderboom et al. describe the quantitative performance of an optimized size-exclusion-chromatography-based isolation method for proteomic analysis of plasma-derived EVs. As proof of concept, this method is applied to detect changes in EV-associated protein abundance after an acute high-intensity aerobic and low-intensity resistance exercise.
Databáze: OpenAIRE
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