Magnetic particle-based immunoprecipitation of nanoscale extracellular vesicles from biofluids

Abstract

Analysis of nanoscale extracellular vesicles (nsEVs) present in blood, cell culture media, and other biofluids has shown tremendous promise in enabling the development of noninvasive blood-based clinical diagnostic tests, predicting and monitoring the efficacy of treatment programs, and providing molecular level insights into pathology that can enlighten new drug targets in the contexts of health conditions such as cancer and Alzheimer’s Disease (AD). In this chapter, we present methods for using magnetic particle-based immunoprecipitation to enrich highly purified populations of nsEVs directly from plasma, serum, and other biofluids. These methods enable downstream analysis of nsEV protein and nucleic acid constituents in the contexts of both global omics profiling and quantification of individual protein or nucleic acid species of interest. Additionally, these methods allow the researcher to either enrich total nsEV populations or enrich nsEVs derived from a particular tissue type from the overall nsEV population. The methods described here are compatible with parallel processing of dozens of biofluid samples and can be valuable tools for enabling nsEV analyses that have high translational relevance in the development of both novel therapeutics and noninvasive diagnostic assays.