Facile preparation of isolation columns filled with integral hybrid materials for efficient isolation of...

Extracellular vesicles (EVs) and their anomalously altered cargoes represent a promising avenue for clinical diagnostics and prognostics. A critical challenge in EV research is the efficient isolation of these vesicles from complex biological samples with high recovery and purity. Although various of materials have good performance in EV isolation, these materials focus on the nanomaterials, which require multiple solution transfer steps in their use process. It will inevitably lead to sample loss, and is difficult to combine with online sample processing methods.

In this study, we introduce a novel isolation column for isolation of EVs, termed EvBHM, which leverages a bi-functional hybrid monolith and a polyethylene (PE) sieve plate. This design integrates the membrane insertion of distearoyl phospholipid ethanolamine (DSPE) with metal affinity chromatography (MAC), utilizing the interaction between titanium ions and the phospholipid membrane of EVs. The PE sieve plate serves as a robust support for the pore structure. This method provides a straightforward and user-friendly approach to prepare the isolation column, which demonstrates superior enrichment efficiency for EVs from microliter of cell culture media or plasma, ensuring minimal sample loss and high purity. Consequently, 37 up-regulated and 91 down-regulated proteins of plasma in colorectal cancer (CRC) patients are found over the health donors, and serval of them are associated with the occurrence and development of CRC.

This method provides a straightforward and user-friendly approach to prepare of the isolation column, which demonstrates superior enrichment efficiency for EVs from microliter of cell culture media or serum as low as 10 μL, ensuring minimal sample loss and high purity. The results suggest this isolated method based on EvBHM isolation column is a promising strategy to search biomarkers for early diagnosis of cancers.