Acoustochromatography based on low-cost polymer columns enables rapid extracellular vesicle proteome analysis from 1 µL blood plasma

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 ≥150 µL 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 minutes - significantly faster than current gold-standard methods. A novel ultrasound-driven release mode increased recovery by up to two orders of magnitude compared to the previously reported acoustofluidic chromatography method. Comprehensive proteomic analysis confirmed highly pure EV fractions after acoustofluidic enrichment. Apolipoproteins were depleted by >99.6%, below levels reported for other methods, lifting 712 mostly EV associated proteins above background as compared to neat plasma. This low-cost, scalable platform unlocks proteomic profiling of ultra-low-volume samples, paving the way for rapid and clinically compatible EV biomarker diagnostics.