Multinodal Acoustic Trapping Enables High Capacity and High Throughput Enrichment of Extracellular Vesicles and Microparticles in miRNA and MS Proteomics Studies.

Axel Broman,Andreas Lenshof,Mikael Evander,Lotta Happonen,Anson Ku,Johan Malmström,Thomas Laurell

doi:10.1021/acs.analchem.0c04772

Abstract

We report a new design of an acoustophoretic trapping device with significantly increased capacity and throughput, compared to current commercial acoustic trapping systems. Acoustic trapping enables nanoparticle and extracellular vesicle (EV) enrichment without ultracentrifugation. Current commercial acoustic trapping technology uses an acoustic single-node resonance and typically operates at flow rates <50 μL/min, which limits the processing of the larger samples. Here, we use a larger capillary that supports an acoustic multinode resonance, which increased the seed particle capacity 40 times and throughput 25–40 times compared to single-node systems. The resulting increase in capacity and throughput was demonstrated by isolation of nanogram amounts of microRNA from acoustically trapped urinary EVs within 10 min. Additionally, the improved trapping performance enabled isolation of extracellular vesicles for downstream mass spectrometry analysis. This was demonstrated by the differential protein abundance profiling of urine samples (1–3 mL), derived from the non-trapped versus trapped urine samples.

Highlights

Extracellular vesicles (EVs) are small, membrane-enclosed particles that are released by cells and contain a wide range of bioactive molecules
After the trapping performance had been evaluated with 500 nm PS beads, we investigated the potential for trapping extracellular vesicles from the urine samples, using the optimized settings from above
We conclude that the sample preparation does not introduce a large increase in relative standard deviation (RSD), which is an important aspect for future quantitative proteomics comparisons for trapped EVs

Summary

Introduction

Extracellular vesicles (EVs) are small, membrane-enclosed particles that are released by cells and contain a wide range of bioactive molecules. The trapping efficiency of 500 nm polystyrene particles (PS) was measured at five different flow rates (100, 200, 500, 1000, and 2000 μL/min). Different volumes of urine (1, 2, or 3 mL) were run through the trap at a flow rate of 500 μL/min to capture EVs. The trap was rinsed with 1 mL

Results

Conclusion