Isolation of Blood-Derived Extracellular Vesicles for Disease Diagnosis: Methods, Challenges, and Applications

Extracellular vesicles (EVs) are a family of cell-derived membrane vesicles found in almost all body fluids. Blood-derived EVs are particularly attractive for clinical diagnostics due to their accessibility and systemic representation. However, the intrinsic complexity of EVs and the abundance of non-EV particles in blood have hindered the establishment of a standardized isolation method. Meanwhile, research on EV isolation is rapidly evolving, and there is a clear need for reviews that synthesize the recent advances in this field, particularly those focusing on blood-derived EVs. In this review, we provide a comprehensive overview of both established and emerging isolation techniques, summarizing their advantages, limitations, and applications reported over the past years, to provide methodological insights and guidance for future isolation studies. • Overview of EV isolation methods from blood samples • Comparison of established and emerging EV techniques • Pros and cons of current EV isolation strategies • Guidance for future EV method development and standardization