抄録
Extracellular vesicles (EVs) carry various informative components, including signaling proteins, transcriptional regulators, lipids, and nucleic acids. EVs have shown great promise as pharmaceutical–targeting vesicles and have attracted the attention of researchers in the fields of biological and medical science because of their importance as diagnostic and prognostic markers. However, the detection and isolation of EVs in cell–cultured media remain challenging. In the present study, we used curvature–sensing peptides to develop a simple and rapid method for in–situ vesicle detection in cultured media without EV–purification steps and applied this method for rapid screening and identification of genes involved in bacterial extracellular vesicle production. Also we developed a novel methodology, i.e., an EV catch–and–release isolation system (EV–CaRiS) using a net–charge invertible curvature–sensing peptide (NIC). NIC was newly designed to reversibly capture and release EVs in a pH–dependent manner. NIC allowed us to achieve reproducible EV isolation from three human cell lines and single–particle imaging of EVs containing the ubiquitous exosome markers CD63 and CD81. EV–CaRiS was demonstrated as a simple and convenient methodology for EV isolation.
