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J. Extracell. Vesicles, Jul 2024, 13 (7) | https://doi.org/10.1002/jev2.12479 Extracellular vesicle isolation and counting system (EVics) based on simultaneous tandem tangential flow filtration and large field-of-view light scattering

Ju-Hyun Bae¹, Chan-Hyeong Lee¹, Dokyung Jung¹, Kyungmoo Yea^2,3, Byoung-Joon Song⁴, Hakho Lee^5,6,Moon-Chang Baek¹

¹Department of Molecular Medicine, CMRI, Exosome Convergence Research Center (ECRC), School of Medicine, Kyungpook National University, Daegu, Republic of Korea

²Department of New Biology, DGIST, Daegu, Republic of Korea

³New Biology Research Center, DGIST, Daegu, Republic of Korea

⁴Section of Molecular Pharmacology and Toxicology, Laboratory of Membrane Biochemistry and Biophysics, National Institute on Alcohol Abuse and Alcoholism, NIH, Bethesda, Maryland, USA

⁵Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA

⁶Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA

Correspondence : Moon-Chang Baek

Abstract

Although the isolation and counting of small extracellular vesicles (sEVs) are essential steps in sEV research, an integrated method with scalability and efficiency has not been developed. Here, we present a scalable and ready-to-use extracellular vesicle (EV) isolation and counting system (EVics) that simultaneously allows isolation and counting in one system. This novel system consists of (i) EVi, a simultaneous tandem tangential flow filtration (TFF)-based EV isolation component by applying two different pore-size TFF filters, and (ii) EVc, an EV counting component using light scattering that captures a large field-of-view (FOV). EVi efficiently isolated 50-200 nm-size sEVs from 15 µL to 2 L samples, outperforming the current state-of-the-art devices in purity and speed. EVc with a large FOV efficiently counted isolated sEVs. EVics enabled early observations of sEV secretion in various cell lines and reduced the cost of evaluating the inhibitory effect of sEV inhibitors by 20-fold. Using EVics, sEVs concentrations and sEV PD-L1 were monitored in a 23-day cancer mouse model, and 160 clinical samples were prepared and successfully applied to diagnosis. These results demonstrate that EVics could become an innovative system for novel findings in basic and applied studies in sEV research.

논문정보

형식Research article
게재일2024년 07월 (BRIC 등록일 2024-07-15)
연구진국내 연구진
분야 의약학 > 분자세포의학

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