Surface Double Dendritic Magnetic Microfibrils for Rapid Isolation and Proteomic Profiling of Extracellular Vesicles from Microliters of Biofluids.

Abstract

The extracellular vesicles (EVs) are crucial for intercellular communication, and their proteomic analysis offers significant insights into their functions, although rapid and efficient analysis in trace biofluids is challenging due to their low abundance and potential protein loss. This study developed functionalized double dendritic magnetic microfibrils (fDDMMs) for efficient isolation and proteomic analysis of EVs from microliter biofluids. The fDDMMs possess dendritic mesoporous silica shell and magnetic Fe₃O₄ core, with bifunctional groups, Ti ions and R8 cell-penetrating peptide, grafted on the surface by dendritic molecules for enhanced EV capture. The multifunctional properties, including dynamic magnetic mixing and accelerated protein digestion, streamline the proteomic sample preparation process. The results demonstrated that fDDMMs enabled the rapid batch separation and proteomic sample preparation of EVs from 1 μL of plasma samples and 100 μL of tumor organoid culture medium. The rapid EV isolation and proteomic profiling approach holds great potential for liquid biopsy and personalized medicine with tiny clinic biofluids.