Rapid and efficient isolation platform for plasma extracellular vesicles: EV-FISHER.

Autor: Pan WL; Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China.; Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, China., Feng JJ; Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China., Luo TT; Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China., Tan Y; Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, China., Situ B; Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China., Nieuwland R; Laboratory of Experimental Clinical Chemistry, Vesicle Observation Centre, Amsterdam University Medical Center, Amsterdam, The Netherlands., Guo JY; Breast Center, Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, China., Liu CC; Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China., Zhang H; Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China., Chen J; Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China., Zhang WH; College of Chemistry, Chemical Engineering and Materials Science, Soochow University, Suzhou, China., Chen J; Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, China., Chen XH; Department of Clinical Laboratory, Liuzhou Municipal Liutie Central Hospital, Liuzhou, China., Chen HY; Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, China., Zheng L; Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China., Chen JX; Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, China., Li B; Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, China.
Jazyk: angličtina
Zdroj: Journal of extracellular vesicles [J Extracell Vesicles] 2022 Nov; Vol. 11 (11), pp. e12281.
DOI: 10.1002/jev2.12281
Abstrakt: Extracellular vesicles (EVs) have found diverse applications in clinical theranostics. However, the current techniques to isolate plasma EVs suffer from burdensome procedures and limited yield. Herein, we report a rapid and efficient EV isolation platform, namely, EV-FISHER, constructed from the metal-organic framework featuring cleavable lipid probes (PO 4 3- -spacer-DNA-cholesterol, PSDC). The EV-FISHER baits EVs from plasma by cholesterol and separates them with an ordinary centrifuge. The captured EVs could be released and collected upon subsequent cleavage of PSDC by deoxyribonuclease I. We conclude that EV-FISHER dramatically outperforms the ultracentrifugation (UC) in terms of time (∼40 min vs. 240 min), isolation efficiency (74.2% vs. 18.1%), and isolation requirement (12,800 g vs. 135,000 g). In addition to the stable performance in plasma, EV-FISHER also exhibited excellent compatibility with downstream single-EV flow cytometry, enabling the identification of glypican-1 (GPC-1) EVs for early diagnosis, clinical stages differentiation, and therapeutic efficacy evaluation in breast cancer cohorts. This work portrays an efficient strategy to isolate EVs from complicated biological fluids with promising potential to facilitate EVs-based theranostics.
(© 2022 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.)
Databáze: MEDLINE
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