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Earlier designated as the “garbage bags” used by the biological systems to remove unnecessary biomolecules of the cells, they have recently gained popularity as secreted vesicles pivotal for intracellular and intercellular information transfer. Studies have shown the presence of essential RNA and protein cargos in these small vesicles (30-150 nm). The interest in exosome studies have exponentially grown as they can be manoeuvred as minimally diagnostic tools in order to understand biological functions. Proteomics of exosomes are considered the biological fingerprints as they replicate the properties of the parental cell from where they are originated. As a mobile container of vital biomolecules (specialized proteins and RNAs) exosomes are crucial for antigen- presentation, cell-cell communication, waste management, translocation of biomolecules and coagulation.
Moreover, recent evidences have reflected the role of exosomes as a non-invasive diagnostic marker to understand pathological conditions as well as connect anterograde or retrograde communications.
Critical study of isolation, characterization and analysis of exosomes and their RNA and protein content can provide an in-depth probing and information in terms of function. Characterization of exosomes can be done by using metabolic incorporation of labelled/modified nucleic acid such as ethynyl uridine or methionine analogs such as homopropargylglycine. The crucial part of the study is to get a pure fraction of exosomes without any subcellular contamination. Hence, various methods have been developed for obtaining pure exosomal fraction.
The most common approach used for the vesicular isolation is ultracentrifugation- based purifications, but this allows infiltration of non-vesicular components of the cellular system, which becomes challenging for consistent diagnosis from samples such as serum, urine or tissue. The other approach is using electron microscopy to get the contaminants information; however this is not a quantitative approach.