Large-scale heparin-based bind-and-elute chromatography identifies two biologically distinct populations of extracellular vesicles
- PMID: 37272197
- PMCID: PMC10240191
- DOI: 10.1002/jev2.12327
Large-scale heparin-based bind-and-elute chromatography identifies two biologically distinct populations of extracellular vesicles
Abstract
Purifying extracellular vesicles (EVs) has been challenging because EVs are heterogeneous in cargo yet share similar sizes and densities. Most surface marker-based affinity separation methods are limited to research or diagnostic scales. We report that heparin chromatography can separate purified EVs into two distinct subpopulations as ascertained by MS/MS: a non-heparin-binding (NHB) fraction that contains classical EV markers such as tetraspanins and a heparin-binding (HB) fraction enriched in fibronectins and histones. Both fractions were similarly fusogenic but induced different transcriptional responses in endothelial cells. While EVs that were purified by conventional, non-affinity methods alone induced ERK1/2 phosphorylation and Ki67, the NHB fraction did not. This result suggests heparin chromatography as an additional novel fractionation step that is inherently scalable, does not lead to loss of material, and separates inflammatory and pyrogenic EVs from unreactive EVs, which will improve clinical applications.
Keywords: exomere; exosome; extracellular particles; extracellular vesicles; fibronectin; heparin.
© 2023 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.
Conflict of interest statement
The authors declare no conflicts of interest.
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