Chromatography and its hyphenation to mass spectrometry for extracellular vesicle analysis.

摘要: Extracellular vesicles (EVs), such as exosomes, microvesicles and apoptotic bodies are released by cells, both under physiological pathological conditions. EVs can participate in a novel type of intercellular communication deliver cargo nucleic acids, proteins lipids near or to distant host cells. EV research is proceeding at fast pace; now they start appear promising therapeutic targets, diagnostic tools drug delivery systems. Isolation analysis prerequisites for understanding their biological roles clinical exploitation. In this process chromatography mass spectrometry (MS)-based strategies rapidly gaining importance; reviewed the present communication. purification mostly performed ultracentrifugation present. Chromatography-based ground, among which affinity size exclusion (SEC) particularly strong contenders. Their major advantages relative simplicity, robustness throughput. Affinity has added advantage separating subtypes based on molecular recognition surface motifs. SEC that isolated may retain activity. typically small amounts, therefore high sensitivity required analysis. Study content (all compounds beside acids) predominantly liquid tandem (LC-MS/MS) The chromatographic separation reverse phase, nanoscale, ultra performance LC technique. MS relying nano-electrospray ionization MS/MS provides sensitivity, selectivity resolution, so thousand(s) be detected/identified/quantified sample. Beside protein identification, quantitation characterization post-translational modifications (PTMs), like glycosylation phosphorylation becoming feasible increasingly important. Along with conventional LC-MS/MS, other approaches hyphenated importance characterization. Hydrophilic interaction used characterize PTMs; LC-inductively coupled plasma/MS identify metal containing molecules; while gas chromatography-MS analyze some metabolites.