Abstract
A method for exosome isolation from human plasma was developed for rapid, high-throughput processing of plasma specimens obtained from patients with cancer. This method removes the bulk of plasma proteins associated with exosomes and can be used for comparative examinations of exosomes and their content in serial specimens of patients’ plasma, allowing for monitoring changes in exosome numbers, profiles, and functions in the course of cancer progression or during therapy. The plasma-derived exosomes can be recovered in quantities sufficient for the characterization of their morphology by transmission electron microscopy (TEM), size and concentration by qNano, protein/lipid ratios, nucleic acid extraction, molecular profiling by Western blots or immune arrays, and functional assays.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Thery C, Amigorena S, Raposo G, Clayton A (2006) Unit 3.22 Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol. doi:10.1002/0471143030.cb0322s30
Hong CS, Funk S, Muller L, Boyiadzis M, Whiteside TL (2016) Isolation of biologically active and morphologically intact exosomes from plasma of patients with cancer. J Extracell Vesicles 5:29289
Taylor DD, Chou IN, Black PH (1983) Isolation of plasma membrane fragments from cultured murine melanoma cells. Biochem Biophys Res Commun 113(2):470–476
Muller L, Hong CS, Stolz DB, Watkins SC, Whiteside TL (2014) Isolation of biologically-active exosomes from human plasma. J Immunol Methods 411:55–65
Saze Z, Schuler PJ, Hong CS, Cheng D, Jackson EK, Whiteside TL (2013) Adenosine production by human B cells and B cell-mediated suppression of activated T cells. Blood 122:9–18
Acknowledgments
The development of the method was in part supported by NIH grants RO-1CA168628 and R21-CA205644 to TLW.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Hong, CS., Funk, S., Whiteside, T.L. (2017). Isolation of Biologically Active Exosomes from Plasma of Patients with Cancer. In: Fortina, P., Londin, E., Park, J., Kricka, L. (eds) Acute Myeloid Leukemia. Methods in Molecular Biology, vol 1633. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7142-8_16
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7142-8_16
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7140-4
Online ISBN: 978-1-4939-7142-8
eBook Packages: Springer Protocols