Isolation and adsorption of extracellular vesicles using molecularly imprinted cryogel membranes

Breast cancer is one of the most significant malignancies worldwide, representing one out of every eight cancers diagnosed in women and affecting one in four women across all ages. Early diagnosis is essential for improving survival and overall quality of life, yet limitations such as the lack of affordable traditional tests and technological or biological barriers often delay timely detection. Extracellular vesicles (EVs), which facilitate information exchange among cancer cells, hold strong potential as primary biomarker for cancer detection. However, the lack of simple, reliable, and reproducible platforms for EV isolation and detection remains a major challenge.

In this study, we focus on the selective isolation of EVs cultured and collected on a microfluidic chip engineered to mimic the cancer microenvironment, using molecularly imprinted cryogel membranes. Optimal conditions for EV adsorption onto these membranes were systematically identified in a batch system using a rotator. The resulting cryogel-based platform demonstrated a high adsorption capacity of 1075 particles per gram of cryogel at pH 5.0. The membranes also showed excellent regeneration performance following EV desorption with 1 M NaCl, supporting sustained operational stability. Strong alignment with the Langmuir adsorption model and validation through HPLC analyses provide both mechanistic insight and analytical confidence, positioning this approach as a robust and reproducible alternative to conventional EV isolation strategies.

Overall, these findings introduce a novel, scalable cryogel-based platform for EV purification, offering high reliability, quantifiability, and reusability. The system's solid analytical performance and compatibility with downstream processing highlight its broad potential in biochemical, diagnostic, and EV-based research and technologies.