Enrichment of extracellular vesicles using Mag-Net for the analysis of the plasma proteome

Christine C. Wu (), Kristine A. Tsantilas, Jea Park, Deanna Plubell, Justin A. Sanders, Previn Naicker, Ireshyn Govender, Sindisiwe Buthelezi, Stoyan Stoychev, Justin Jordaan, Gennifer Merrihew, Eric Huang, Edward D. Parker, Michael Riffle, Andrew N. Hoofnagle, William S. Noble, Kathleen L. Poston, Thomas J. Montine and Michael J. MacCoss ()
Additional contact information
Christine C. Wu: University of Washington
Kristine A. Tsantilas: University of Washington
Jea Park: University of Washington
Deanna Plubell: University of Washington
Justin A. Sanders: University of Washington
Previn Naicker: CSIR
Ireshyn Govender: ReSyn Biosciences
Sindisiwe Buthelezi: CSIR
Stoyan Stoychev: ReSyn Biosciences
Justin Jordaan: ReSyn Biosciences
Gennifer Merrihew: University of Washington
Eric Huang: University of Washington
Edward D. Parker: University of Washington
Michael Riffle: University of Washington
Andrew N. Hoofnagle: University of Washington
William S. Noble: University of Washington
Kathleen L. Poston: Stanford University
Thomas J. Montine: Stanford University
Michael J. MacCoss: University of Washington

Nature Communications, 2025, vol. 16, issue 1, 1-15

Abstract: Abstract Extracellular vesicles (EVs) in plasma are composed of exosomes, microvesicles, and apoptotic bodies. We report a plasma EV enrichment strategy using magnetic beads called Mag-Net. Proteomic interrogation of this plasma EV fraction enables the detection of proteins that are beyond the dynamic range of liquid chromatography-mass spectrometry of unfractionated plasma. Mag-Net is robust, reproducible, inexpensive, and requires 37,000 peptides from >4,000 proteins. Using Mag-Net on a pilot cohort of patients with neurodegenerative disease and healthy controls, we find 204 proteins that differentiate (q-value

Date: 2025
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DOI: 10.1038/s41467-025-60595-7

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