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Theranostics, 2019. 9(7), 1851-1863, DOI: https://doi.org/10.7150/thno.32438
Fully Automated, Label-Free Isolation of Extracellular Vesicles from Whole Blood for Cancer Diagnosis and Monitoring
Sunkara Vijaya, Kim Chi-Ju;Park Juhee;Woo Hyun-Kyung;Kim Dongyoung;Ha Hong Koo;Kim Mi-Hyun;Son Youlim;Kim Jae-Ryong;Cho Yoon-Kyoung
Extracellular vesicles (EVs) that circulate in body fluids possess significant potential for disease diagnosis.Their use in clinical settings, however, has been limited owing to lack of simple and robust isolationmethods. To rectify this problem, a centrifugal device for automatic, fast, and efficient isolation of EVsfrom whole-blood, called Exodisc-B is presented in this paper.Methods: The device comprises a built-in chamber to facilitate plasma separation and two nanoporousfilters—one for removing larger particles and the other for enriching EVs. The performance of the devicein comparison to ultracentrifugation (UC) was evaluated by analyzing the yield, purity, protein and RNAcontent of the isolated EVs. Additionally, the EV protein marker expressions were measured by ELISAand statistically analyzed to differentiate prostate cancer patients from healthy donors.Results: Compared with the UC technique, the proposed device is capable of isolating at least an orderof magnitude higher number of EVs with about 30-fold higher mRNA count within 40 min. SandwichELISA of EV-specific membrane proteins—CD9-CD81—confirmed that Exodisc-B can isolate EVs from avolume of whole blood as low as 30 μL with a capture efficiency exceeding 75%. The device also facilitatestemporal monitoring of tumor progression within live mouse xenograft models over a period of 13weeks while using minimal volumes of weekly collected blood samples. Further, in ELISA analyses ofmultiple cancer-related proteins, such as prostate-specific antigen (PSA), prostate-specific membraneantigen (PSMA), epithelial cell adhesion molecule (EpCAM), epidermal growth factor receptor 1 (EGFR1),and heat shock protein 90 (HSP90), extracted from EVs isolated from human plasma, 43 patients weredifferentiated from 30 healthy donors.Conclusion: The results demonstrated the ability of Exodisc-B to provide a rapid, sensitive, andpoint-of-care-type method for extracting intact EVs from small volumes of clinical blood samples fordisease diagnosis and monitoring.
- DOI: https://doi.org/10.7150/thno.32438
- ISBN or ISSN: 1838-7640
- 본 연구는 질병관리본부 연구개발과제연구비를 지원받아 수행되었습니다.
- This research was supported by a fund by Research of Korea Centers for Disease Control and Prevention.