A pH-Responsive DNA Platform for Gentle Enrichment and Efficient Recovery of Extracellular Vesicles

Extracellular vesicles (EVs) have emerged as crucial mediators of intercellular communication, capable of transporting a wide array of biomolecules. Their exceptional potential as both biomarkers and therapeutic agents has garnered significant attention in biomedical research. However, the broader application of EVs is hampered by substantial challenges associated with current enrichment strategies, which often require sophisticated equipment, involve complex operational procedures, and risk compromising vesicle integrity. In this study, we introduce a novel DNA-based platform, termed SiTc, which integrates intelligent pH-responsive properties for rapid and efficient EVs enrichment, as well as reversible release. Under neutral pH conditions, SiTc facilitates the spontaneous aggregation of nanoscale EVs into microscale clusters, enabling their rapid and gentle isolation by low-speed centrifugation. Furthermore, the SiTc platform is equipped with a reversible, pH-triggered disassembly mechanism that allows for controlled dissociation of EVs-clusters and efficient recovery of intact, individual EVs with high yield. Comparative analyses demonstrate that the SiTc platform outperforms conventional ultracentrifugation methods, particularly in enriching low-abundance EV samples, by delivering superior recovery efficiency, enhanced reproducibility, and scalability. By streamlining the isolation process, while preserving EV integrity, this methodology represents a transformative advancement in EV enrichment, holding promise for accelerating EV-based research and facilitating the clinical translation of EVs.