Exosomes have shown great potential in disease diagnostics and therapeutics. However, current isolation approaches are burdensome and suffer from low speed, yield and purity, limiting basic research and clinical applications. Here, we describe an efficient exosome detection method via the ultrafast-isolation system (EXODUS) that allows automated label-free purification of exosomes from varied biofluids. We obtained the ultra-efficient purification of exosomes by negative pressure oscillation and double coupled harmonic oscillator–enabled membrane vibration. Our two coupled oscillators generate dual-frequency transverse waves on the membranes, enabling EXODUS to outperform other isolation techniques in speed, purity and yield. We demonstrated EXODUS by purifying exosomes from urine samples of 113 patients and validated the practical relevance in exosomal RNA profiling with the high-resolution capability and high-throughput analysis.
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We thank Tongji Hospital in the Tongji Medical College at Huazhong University of Science and Technology for providing clinical samples in this study. The work was primarily supported by research fund provided by the Zhenan Technology City Research Fund, the Zhejiang Provincial and Ministry of Health Research Fund for Medical Sciences (grant no. WKJ-ZJ-1910), the Wenzhou Medical University (grant no. 89218012) and the Wenzhou Institute, University of Chinese Academy of Sciences (grant no. WIBEZD2017006-05).
The authors declare no competing interests.
Peer review information Madhura Mukhopadhyay was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
a, A photo of the EXODUS device, with its cross-sectional illustration. Scale bar, 1 cm. b, Illustration of harmonic oscillations in the EXODUS device for minimizing fouling effect and bio-aggregates. c, The vibration motors generate acoustic streaming at different fluidic levels inside the EXODUS device. Scale bar, 1 cm.
a, An image of the workstation. Scale bar, 10 cm. b, Internal view of the workstation, including (1) autosampler, (2) needle, (3) needle wash site, (4) EXODUS device window, (5) specimen tube, (6) wash buffer A, and (7) wash buffer B. Scale bar, 3 cm. c, The interface to EXODUS device via station: a device slot moves out from inside by pressing the button “Install Device” on the control panel to install EXODUS’s membrane device onto the workstation. d, The architecture of the EXODUS workstation. e, The illustration of the fluidic system in the EXODUS workstation.
Extended Data Fig. 3 Simulations of pressure, flow distribution, and vibration modes of the EXODUS device.
a, COMSOL simulation showing the pressure distribution in a cross-section view of the EXODUS device when applying a −20 kPa negative pressure (NP) from the L-outlet. The sample is injected from the top opening into the sample reservoir. b, Distribution of flow velocity on the membrane from top to bottom. The flow velocity is between 45 and 60 μm/s, with a variation below 25%. c, Simulation of different vibration modes for a clamped circular nanoporous AAO membrane at their representative vibration frequencies: 469.8 Hz (5–700 Hz), 1281.8 Hz (700–1600 Hz), 1861.8 Hz (1600–3000 Hz), 3717.8 Hz (3000–5000 Hz), 5921.8 Hz (5000–8000 Hz), and 10562 Hz (8000–11500 Hz). The integer index m refers to the azimuthal node number. The index n refers to the nth non-trivial zero of the Bessel function. The high-frequency oscillation of the membrane at 6250 Hz by piezoelectric transducer has a (0,4) vibration mode, while the low high-frequency oscillation of the membrane at 200 Hz by vibration motor has a (0,1) vibration mode. The scale ranges from no displacement (dark blue) to maximum displacement (dark red).
a, Comparison of the sample processing times of EXODUS and other methods showing their detailed procedures. For each method, the time cost was basically calculated according to its protocol. b, Size distributions and c amount of the particles obtained by EXODUS and other methods from 10 mL of urine sample aliquots. (n≥5 independent experiments). d, Isolation of exosomes from 10 mL of urine sample aliquots with 20 replications to study the reproducibility of EXODUS. Qubit™ Protein Assay Kits measured the total protein amounts of isolated samples, with an average protein amount of 4.3 μg and a CV of 9.9% over the 20 measurements. e, The concentrations and f size distributions of the particles isolated from different biofluids by EXODUS. (n = 3 independent experiments). The concentration of tear exosomes is presented as particles per centimeter tear collection paper. In c and e, data are presented as mean value ± SD. NTA profiles in b and f are constructed by the average curve (solid line) and error band (shaded area). Source data
a, NTA profiles of EV particles with serious cut-off size ranges of 20–450, 20–100, 100–200, and 200–450 nm. b, The typical TEM images of vesicle subpopulations and the corresponding statistics of size distributions are summarized in c. (20–450 nm: n = 565 independent EVs, 20–100 nm: n = 386 independent EVs, 100–200 nm: n = 46 independent EVs, 200–450 nm: n = 31 independent EVs). Source data
Supplementary Figs. 1–17 and Notes 1–3.
Transverse waves observed at the top surface of liquid in PEI device.
Transverse waves observed in the deep of PEI device.
Transverse waves observed in a PMMA device.
Clinical information of patients
Original western blot scans.
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Chen, Y., Zhu, Q., Cheng, L. et al. Exosome detection via the ultrafast-isolation system: EXODUS. Nat Methods 18, 212–218 (2021). https://doi.org/10.1038/s41592-020-01034-x