Real-Time Isolation and Versatile Detection for Extracellular Vesicles Based on Ordered Porous Layer Interferometry.

Extracellular vesicles (EVs) are progressively becoming novel instruments for clinical therapeutics and liquid biopsies. Due to the complexity of biofluids and the physicochemical properties of EVs, the biological activity, velocity, and efficiency of EV isolation are always unsatisfying. Here, we present a real-time isolation approach of EVs derived from cells and urine using ordered porous layer interferometry with a silica colloidal crystal film as the sensing substrate, achieving efficiency greater than 90%. The online concentration detection function is performed during the isolation process on the basis of its real-time monitoring characteristic. Using membrane protein markers of urine EVs as targets, this technique has a high diagnostic value for liquid biopsy of prostate cancer. Furthermore, we compared multiple EV membrane protein expression and binding dissociation kinetic data from cells and urine. In summary, this multifunctional approach provides a novel strategy for the rapid EVs isolation, concentration detection, drug target screening, and liquid biopsy of various body fluids.