Acoustochromatography Based on Low-Cost Polymer Columns Enables Rapid Extracellular Vesicle Proteome Analysis from 1 μL of Blood Plasma

Abstract Extracellular vesicles (EVs) hold promise as biomarkers for early disease detection, yet their nanoscale size and the protein-rich plasma background make enrichment essential. Conventional approaches such as ultracentrifugation and size-exclusion chromatography require ≥100 μL of input, hours of processing, and are poorly compatible with clinical workflows. We present a low-cost polymer-based acoustochromatography device that enables EV enrichment from 1 μL of blood plasma within 2–10 min─significantly faster than current gold-standard methods. A novel ultrasound-driven release mode increased recovery significantly compared to the previously reported acoustofluidic chromatography method. Comprehensive proteomic analysis confirmed highly pure EV fractions after acoustofluidic enrichment. Apolipoproteins were depleted to below 100 ng/mL, corresponding to >99.6% depletion as compared to neat plasma. This absolute residual concentration is lower than values reported for other EV isolation methods and enabled the detection of an additional 689 mostly EV-associated proteins above the plasma background. This low-cost, scalable platform unlocks proteomic profiling of ultralow-volume samples, paving the way for rapid and clinically compatible EV biomarker diagnostics.