Abstract
The presented procedure allows rapid isolation of extracellular vesicles (EVs) from plasma using size-exclusion chromatography (SEC). Additionally, an approach for reducing the lipid and salt content of the EV isolate in preparation for mass spectrometry (MS)-based proteomic analysis is presented. An example setup for proteomic profiling of the processed samples by nanoflow liquid chromatography coupled to tandem mass spectrometry (nLC-MS/MS) is also presented. Approximately 1000 protein groups in blood plasma-derived EVs can be identified and quantitated following this procedure and using the described instrumentation.
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Acknowledgments
This work was supported by the ASMS 2015 Research Award (ARI), NIH 1R01GM120272-01 (ARI), and the Dana-Farber Cancer Institute/Northeastern University Joint Seed Funding Program in Cancer Drug Development (ARI). The authors thank Dr. Ionita Ghiran (Beth Israel Deaconess Medical Center) for fruitful discussions and his assistance with blood collection.
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Kreimer, S., Ivanov, A.R. (2017). Rapid Isolation of Extracellular Vesicles from Blood Plasma with Size-Exclusion Chromatography Followed by Mass Spectrometry-Based Proteomic Profiling. In: Kuo, W., Jia, S. (eds) Extracellular Vesicles. Methods in Molecular Biology, vol 1660. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7253-1_24
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DOI: https://doi.org/10.1007/978-1-4939-7253-1_24
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