ExoDisc for Urine
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(ExoDisc D20)
The performance of ExoDisc was compared with other commonly used EV-isolation methods. EVs isolated from PC3 cells (ATCC CRL-1435) transfected with AR-FL and AR-V7 were spiked to 4 ml of healthy urine. Isolated EVs were characterized by nanoparticle-tracking analysis (NTA), enzyme-linked immunosorbent assay (ELISA), and polymerase chain reaction (PCR). An ExoDisc provides > 95% recovery of EVs, 3 - 17x higher yield, and > 5x higher purity than the standard ultracentrifugation method.
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High Performance
NTA (Nanoparticle Tracking Analysis)
ExoDisc gives a much higher number of particles over a broad range of sizes.
CD9 concentration is considerably higher when isolated by an ExoDisc than by UC (ultracentrifugation).
Urine samples from prostate cancer patients contain Androgen Receptor (AR) mRNAs in different forms, and an ExoDisc yields about 100 times more AR mRNA concentration which implies the advantage as a diagnostic analysis tool.
High Yield
NTA (Nanoparticle Tracking Analysis)
ELISA CD9/CD81 (sandwich assay)
Capture Efficiency / Western Blotting
The comparison of the number of particles from different methods. ExoDisc results give 4 times more particles than UC (ultracentrifugation).
Not all particles collected are EVs. The particles obtained from ExoDisc include the most amount of particles with EV-specific surface markers (CD9, CD81).
By spiking labeled EVs in the urine, ExoDisc gives the highest capture efficiency using cell line-generated EVs. surpassing the results of UC (ultracentrifugation). The EV internal marker Alix was found much more abundant by western blotting.
High purity
The purity of isolated EVs is commonly quantified by the number of particles per amount of protein.
An ExoDisc shows good purity in comparison.
The performance of an ExoDisc does not degrade after repeated loading of urine volumes, showing a linear increase of isolated particles which cannot be obtained in other methods.
High Reproducibility
The number of particles, EV mount, capture efficiency, and real-time PCR results using the same sample using different ExoDiscs over several weeks give consistent results. The relative abundance of mRNA cargo is reproducible.
References
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Woo, H.K., Sunkara, V. et al. Exodisc for Rapid, Size-Selective, and Efficient Isolation and Analysis of Nanoscale Extracellular Vesicles from Biological Samples. ACS Nano 11, 1360–1370 (2017). https://doi.org/10.1021/acsnano.6b06131
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Woo, H.K., Park, J. et al. Urine-based Liquid Biopsy: Non-invasive and Sensitive AR-V7 Detection in Urinary EVs from Patients with Prostate Cancer. Lab Chip 19, 87-97 (2019). https://doi.org/10.1039/C8LC01185K
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Dong, L., Zieren, R. C., Horie, K. et al. Comprehensive Evaluation of Methods for Small Extracellular Vesicles Separation from Human Plasma, Urine and Cell Culture Medium. J Extracell Vesicles (2020). https://doi.org/10.1002/jev2.12044