TY - JOUR
T1 - A size-exclusion-based approach for purifying extracellular vesicles from human plasma
AU - Vanderboom, Patrick M.
AU - Dasari, Surendra
AU - Ruegsegger, Gregory N.
AU - Pataky, Mark W.
AU - Lucien, Fabrice
AU - Heppelmann, Carrie Jo
AU - Lanza, Ian R.
AU - Nair, K. Sreekumaran
N1 - Funding Information:
Support for this work was provided by National Institutes of Health grants MoTrPAC Chemical Analysis Center grant U24-DK-112326 , CTSA grant number UL1 TR002377 from the National Center for Advancing Translational Sciences , Mayo Clinic Metabolomics Resource Core grants U24-DK-100469 , T32-AG-49672 , R01 AG062859 , and the Dr. Richard F. Emslander Professorship in Endocrinology and Nutrition Research (K.S.N.). The authors thank Dawn Morse, Katherine Klaus, and F.L.-Matteoni for their skillful assistance.
Publisher Copyright:
© 2021 The Authors
PY - 2021/7/26
Y1 - 2021/7/26
N2 - 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.
AB - 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.
KW - bioinformatics
KW - high-intensity exercise
KW - mass spectrometry
KW - size-exclusion chromatography
KW - ultracentrifugation
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U2 - 10.1016/j.crmeth.2021.100055
DO - 10.1016/j.crmeth.2021.100055
M3 - Article
C2 - 34355211
AN - SCOPUS:85126435634
SN - 2667-2375
VL - 1
JO - Cell Reports Methods
JF - Cell Reports Methods
IS - 3
M1 - 100055
ER -