Abstract
Exosomes are the smallest extracellular vesicles present in most of the biological fluids. They are found to play an important role in cell signaling, immune response, tumor metastasis, etc. Studies have shown that these vesicles also have diagnostic and therapeutic roles for which their accurate detection and quantification is essential. Due to the complexity in size and structure of exosomes, even the gold standard methods face challenges. This comprehensive review discusses the various standard methods such as ultracentrifugation, ultrafiltration, size-exclusion chromatography, precipitation, immunoaffinity, and microfluidic technologies for the isolation of exosomes. The principle of isolation of each method is described, as well as their specific advantages and disadvantages. Quantification of exosomes by nanoparticle tracking analysis, flow cytometry, tunable resistive pulse sensing, electron microscopy, dynamic light scattering, and microfluidic devices are also described, along with the applications of exosomes in various biomedical domains.
Highlights
Extracellular vesicles (EVs) are membrane-bound lipid vesicles
Selective quantification of biomarkers in nanovesicles derived from cancerous tissues was performed using microfluidicsbased on liquid biopsy screening tests which improved the accuracy of the results [121]
Apart from being a biomarker, exosomes have shown its role as a therapeutic agent against diseases
Summary
Extracellular vesicles (EVs) are membrane-bound lipid vesicles. They are spherical secretory vesicles with a proteolipid bilayer membrane, their diameters ranging from 30–2000 nm [1]. The complexes combine with ESCRT-III, which interacts with Vps to cleave the necks of the buds to form ILVs. recent evidence suggests that there could be an ESCRT-independent pathway for exosome formation and cargo loading, which uses raft-based lipid microdomains of the plasma membrane and associated proteins like tetraspanins [30]. The protein composition of extracellular vesicles is a good indicator of the subtype of EV, the mode of biogenesis and release, and the original cell type Regardless of these factors, all exosomes contain some common proteins, for instance, heat shock protein 84 (Hsp84), tumor susceptibility gene 101 (TSG101) and Alix, for transport mechanism [7]. Several studies show that RNA can be transferred within exosomes from cell to cell [41, 42], the extent to which this transferred RNA is functional in the recipient cells and the extent of their fragmentation and transfer is yet to be determined
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