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Fully Automated, Label-Free Isolation of Extracellular Vesicles from Whole Blood for Cancer Diagnosis and Monitoring
- Source :
- Theranostics
- Publication Year :
- 2019
- Publisher :
- Ivyspring International Publisher, 2019.
-
Abstract
- 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 isolation methods. To rectify this problem, a centrifugal device for automatic, fast, and efficient isolation of EVs from whole-blood, called Exodisc-B is presented in this paper. Methods: The device comprises a built-in chamber to facilitate plasma separation and two nanoporous filters—one for removing larger particles and the other for enriching EVs. The performance of the device in comparison to ultracentrifugation (UC) was evaluated by analyzing the yield, purity, protein and RNA content of the isolated EVs. Additionally, the EV protein marker expressions were measured by ELISA and 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 order of magnitude higher number of EVs with about 30-fold higher mRNA count within 40 min. Sandwich ELISA of EV-specific membrane proteins—CD9-CD81—confirmed that Exodisc-B can isolate EVs from a volume of whole blood as low as 30 µL with a capture efficiency exceeding 75%. The device also facilitates temporal monitoring of tumor progression within live mouse xenograft models over a period of 13 weeks while using minimal volumes of weekly collected blood samples. Further, in ELISA analyses of multiple cancer-related proteins, such as prostate-specific antigen (PSA), prostate-specific membrane antigen (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 were differentiated from 30 healthy donors. Conclusion: The results demonstrated the ability of Exodisc-B to provide a rapid, sensitive, and point-of-care-type method for extracting intact EVs from small volumes of clinical blood samples for disease diagnosis and monitoring.
- Subjects :
- Male
Mice, Nude
Medicine (miscellaneous)
Blood Donors
Enzyme-Linked Immunosorbent Assay
Extracellular Vesicles
Mice
03 medical and health sciences
Prostate cancer
chemistry.chemical_compound
0302 clinical medicine
Antigen
Heat shock protein
Biomarkers, Tumor
medicine
Animals
Humans
lab-on-a-disc
RNA, Messenger
Liquid biopsy
Pharmacology, Toxicology and Pharmaceutics (miscellaneous)
030304 developmental biology
Whole blood
Automation, Laboratory
size-based filtration
0303 health sciences
Messenger RNA
liquid biopsy
Chemistry
Prostatic Neoplasms
Epithelial cell adhesion molecule
medicine.disease
Molecular biology
Tumor progression
030220 oncology & carcinogenesis
ELISA
Ultracentrifugation
Research Paper
Subjects
Details
- ISSN :
- 18387640
- Volume :
- 9
- Database :
- OpenAIRE
- Journal :
- Theranostics
- Accession number :
- edsair.doi.dedup.....dd82d2f28de4ba2cddcefb2dfa7fe197