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나노의학 연구단
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Integrated Dual-Mode Chromatography to Enrich Extracellular Vesicles from Plasma

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Title
Integrated Dual-Mode Chromatography to Enrich Extracellular Vesicles from Plasma
Author(s)
Jan Van Deun; Ala Jo; Huiyan Li; Hsing-Ying Lin; Ralph Weissleder; Hyungsoon Im; Hakho Lee
Publication Date
2020-12
Journal
Advanced Biosystems, v.4, no.12
Publisher
WILEY-V C H VERLAG GMB
Abstract
Purifying extracellular vesicles (EVs) from complex biological fluids is a critical step in analyzing EVs molecularly. Plasma lipoprotein particles (LPPs) are a significant confounding factor as they outnumber EVs >104-fold. Given their overlap in size, LPPs cannot be completely removed using standard size-exclusion chromatography. Density-based separation of LPPs can be applied but is impractical for routine use in clinical research and practice. Here a new separation approach, known as dual-mode chromatography (DMC), capable of enriching plasma EVs, and depleting LPPs is reported. DMC conveniently integrates two orthogonal separation steps in a single column device: i) size exclusion to remove high-density lipoproteins (HDLs) that are smaller than EVs; and ii) cation exchange to clear positively charged ApoB100-containing LPPs, mostly (very) low-density lipoproteins (V)LDLs, from negatively charged EVs. The strategy enables DMC to deplete most LPPs (>97% of HDLs and >99% of (V)LDLs) from human plasma, while retaining EVs (>30% of input). Furthermore, the two-in-one operation is fast (15 min per sample) and equipment-free. With abundant LPPs removed, DMC-prepared samples facilitate EV identification in imaging analyses and improve the accuracy for EV protein analysis
URI
https://pr.ibs.re.kr/handle/8788114/9013
DOI
10.1002/adbi.201900310
ISSN
2366-7478
Appears in Collections:
Center for Nanomedicine (나노의학 연구단) > 1. Journal Papers (저널논문)
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