Purification of High Yield Extracellular Vesicle Preparations Away from Virus

11.4K Views

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00:07 min

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September 12th, 2019

DOI :

10.3791/59876-v

September 12th, 2019

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Catherine DeMarino*¹, Robert A. Barclay*¹, Michelle L. Pleet¹, Daniel O. Pinto¹, Heather Branscome¹^,², Siddhartha Paul³, Benjamin Lepene⁴, Nazira El-Hage⁵, Fatah Kashanchi¹

¹Laboratory of Molecular Virology, School of Systems Biology, George Mason University, ²American Type Culture Collection (ATCC), ³ATCC Cell Systems, ⁴Ceres Nanosciences, Inc, ⁵Department of Immunology and Nano-medicine, Herbert Wertheim College of Medicine, Florida International University

Summary

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Extracellular Vesicles

Purification Protocol

Virion Separation

Ultracentrifugation

Nanoparticle Preparation

PEG Precipitation

Density Gradient

Iodixanol Gradient

Downstream Analysis

Viral Infections

Chapters in this video

0:04

Title

1:00

Filtration and Precipitation of Extracellular Vesicles

2:25

Construction of a Density Gradient and Enrichment of EV Fractions

4:06

Preparation of Nanoparticle Pellet for Downstream Assay

5:05

Results: Increased EV Yield and Characterization of EVs

6:09

Conclusion

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