Porphyrin-based micro-nano assemblies combine with a well plate assay for capturing and molecular profiling of extracellular vesicles.

Extracellular vesicles (EVs) have displayed promising features as biomarkers for early cancer diagnoses. Inorganic materials such as ZnO-based nanowire arrays have been developed to capture EVs via electrostatic interactions, being a simple method without requiring immunoaffinity capture. However, ZnO-based nanowire arrays have high hardness, which results in lower EV capture efficiencies. Still, to date, organic nano-micro materials have not been used to capture EVs. Herein, polyethyleneimine (PEI)-modified 5,10,15,20-tetrakis(4-(hydroxyl)phenyl)porphyrin (THPP) assemblies were prepared via an evaporation-induced self-assembly method. The obtained THPP/PEI assemblies have a positive surface potential under physiological conditions. Therefore, THPP/PEI assemblies were used to capture EVs derived from cancer cells via electrostatic interaction, reaching 60.6% EV capture efficiency. On the other hand, in conventional methods, analysis of membrane proteins on EVs needs an additional test after isolation. Herein, we integrated THPP/PEI assemblies and a 24-well plate assay into a measurement system that was suitable for both EV capture and profiling membrane proteins of EVs to search for EV markers from the cancer cell medium and urine samples. Using this measurement system, we demonstrated that the membrane protein expression ratio of CD147/CD9 of EVs derived from colon cancer cells was 2.25-fold higher than that from normal colon cells with a p-value less than 0.01. Our results indicate that the integrated measurement system allows users not only to capture EVs effectively but also to profile membrane proteins on EVs. We anticipate that the integrated measurement system will be a powerful tool for early cancer diagnoses.