Tissue-derived extracellular vesicles and their use as diagnostics

摘要

A method of isolating extracellular vesicles (EVs) and identifying membrane proteins from human tissues comprising isolating EVs, treating the EVs with a solution having a pH in a range of 9 to 14 to obtain membranes, isolating membranes, identifying proteins on the membranes using mass spectrometry and isolating EVs with such membrane proteins to identify disease-specific proteome, RNA cargo or DNA cargo. Also claimed are methods for diagnosing diseases using tissue or disease-specific membrane proteins on EVs by capturing EVs using an epitope specific binder and detecting EVs using an epitope specific binder or detecting nucleic acids on or in the captured extracellular vesicles. Further claimed is a method for diagnosing diseases using tissue or disease-specific membrane proteins on EVs comprising adding an epitope-specific binder against a tissue or disease-specific membrane protein and a lipid-biotin conjugate to a sample and detecting a tissue-specific membrane protein by using alpha LISA donor and acceptor beads. Also claimed is a kit comprising an epitope-specific binder against tissue or disease-specific membrane proteins and a disease associated marker detection agent.