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| Main Authors: | , , , , , , , , , , , , |
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| Format: | Artículo científico |
| Language: | en |
| Published: |
Scientific reports
2026
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| Online Access: | https://pubmed.ncbi.nlm.nih.gov/42162070/ |
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Table of Contents:
- Hydrophobic interaction chromatography resolves extracellular vesicle fractions with distinct lipidomic signatures. Młynarczyk, Michał Więckowska, Wiktoria Belka, Mariusz Ewonde Ewonde, Raphael Mantej, Jagoda Klimczuk, Mikołaj Gajdowska, Felicja Matinha-Cardoso, Jorge Tamagnini, Paula Gutowska-Owsiak, Danuta Oliveira, Paulo Eeltink, Sebastiaan Hewelt-Belka, Weronika Current extracellular vesicle (EV) isolation workflows are dominated by size- and density-based approaches, which provide limited insight into surface chemical properties of vesicular particles. Here, we report a hydrophobic interaction chromatography (HIC) workflow for resolving EV fractions along differences in membrane interfacial hydrophobicity. Using a commercially available HIC column, reproducible fractionation of EV samples was achieved, yielding discrete fractions that differed in retention behaviour and lipid composition. Transmission electron microscopy (TEM) showed vesicle-like nanoparticles with EV-consistent morphology, indicating preservation of vesicle integrity during HIC. Lipidomic profiling by reversed-phase liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RP-LC-Q-TOF-MS) revealed fraction-specific differences in lipid composition. Importantly, we demonstrate the feasibility of direct injection of pre-cleaned biofluids onto the HIC column, enabling fractionation of nanoparticle populations containing EVs without prior ultracentrifugation. Furthermore, an operational interfacial hydrophobicity index derived from lipidomic data showed a clear correlation with HIC retention, providing an orthogonal compositional descriptor consistent with the proposed fractionation mechanism. Together, this hydrophobicity-based strategy introduces a previously unexplored physicochemical dimension to EV analysis, revealing chemically structured heterogeneity that is not accessible using conventional separation strategies. The workflow provides a practical framework for fractionating EVs in a manner directly relevant to lipidomic profiling and studies of EV membrane chemistry.