Identification of early stage malignancy changes in the structural and biophysical proporties of exosomes by atomic force microscopy (AFM)-based nano-mechanical measurements

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We applied 3D-AFM to assess the structural and nanomechanical properties of exosomes released from different types of cells in a physiologicallyrelevant environment. We established A method for studying the interaction of molecules exposed on the exosomes by using AFM. Exploring the molecule exposed on the native exosome surface will deepen our understanding of the physiological function of exosomes in cancer progression. In addition, we performed AFM measurements to identify the exosomes in heterogeneous extracellular vesicles (EVs) and analysis their biophysical properties. There is no method that can isolate exosomes (EXOs) or microvesicles (MVs) without including other EVs. As a substitute for EXOs, intraluminal vesicles (ILVs), which are the origin of EXOs were isolated from cells. After separating EVs by the anion exchange column, we performed AFM nanomechanical characterization to determine the biophysical properties of exosomes and other EV subtypes.AFM nanomechanical measurements determined the rapture characteristic of EVs. We have found differences in their rapture mechanics, implying difference of membrane structure related to elastic fibers. Mass spectroscopic analyses were performed to determine their protein and lipid compositions.