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Immuno-acoustic trapping for extracellular vesicle subpopulations

Broman, Axel LU ; Havers, Megan LU orcid ; Sattarov, Roman LU and Laurell, Thomas LU (2025) In Scientific Reports 15(1).
Abstract

Extracellular vesicles (EVs) in biofluids are heterogeneous in origin, surface protein expression, and biomolecular cargo. Isolation of specific EV subpopulations may enable studies of biomarkers originating from specific cell phenotypes and disease states. Acoustic trapping is a rapid, automatable EV isolation technique requiring minimal sample preprocessing; however, it lacks the specificity needed for some clinical studies. Combining acoustic trapping with immunoaffinity bead-based isolation could provide access to more specific EV content and broaden the applications of acoustic trapping. Here, we present a novel immuno-acoustic trapping methodology implemented on an automated platform with simple design. EVs were rapidly isolated... (More)

Extracellular vesicles (EVs) in biofluids are heterogeneous in origin, surface protein expression, and biomolecular cargo. Isolation of specific EV subpopulations may enable studies of biomarkers originating from specific cell phenotypes and disease states. Acoustic trapping is a rapid, automatable EV isolation technique requiring minimal sample preprocessing; however, it lacks the specificity needed for some clinical studies. Combining acoustic trapping with immunoaffinity bead-based isolation could provide access to more specific EV content and broaden the applications of acoustic trapping. Here, we present a novel immuno-acoustic trapping methodology implemented on an automated platform with simple design. EVs were rapidly isolated (8 min) from ~ 17 µL blood plasma, generating two EV fractions from a single trapping run: acoustically trapped EVs and EVs bound specifically to anti-CD9 silica beads. We benchmark our technique against a manual (90 min) immunoaffinity isolation with multiple centrifugation washes. Nanoparticle tracking analysis and transmission electron microscopy with CD9+ immunogold labeling confirmed isolation of intact EVs. Quantitative proteomic profiling showed that immuno-acoustic fractions contained proteins more strongly associated with CD9 than both acoustically and immunoaffinity-isolated fractions. This methodology can be adapted to isolate specific EV subpopulations expressing other surface proteins while simultaneously collecting a broad EV population as background reference.

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author
; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Acoustic trapping, Extracellular vesicles, Immuno-acoustic trapping, Immunoaffinity, Proteomics, Subpopulation
in
Scientific Reports
volume
15
issue
1
article number
45805
publisher
Nature Publishing Group
external identifiers
  • scopus:105026398744
  • pmid:41476096
ISSN
2045-2322
DOI
10.1038/s41598-025-33842-6
language
English
LU publication?
yes
id
a2ba56bf-bef9-49ad-a29c-be6b3f7587e6
date added to LUP
2026-02-11 12:56:38
date last changed
2026-06-04 04:21:45
@article{a2ba56bf-bef9-49ad-a29c-be6b3f7587e6,
  abstract     = {{<p>Extracellular vesicles (EVs) in biofluids are heterogeneous in origin, surface protein expression, and biomolecular cargo. Isolation of specific EV subpopulations may enable studies of biomarkers originating from specific cell phenotypes and disease states. Acoustic trapping is a rapid, automatable EV isolation technique requiring minimal sample preprocessing; however, it lacks the specificity needed for some clinical studies. Combining acoustic trapping with immunoaffinity bead-based isolation could provide access to more specific EV content and broaden the applications of acoustic trapping. Here, we present a novel immuno-acoustic trapping methodology implemented on an automated platform with simple design. EVs were rapidly isolated (8 min) from ~ 17 µL blood plasma, generating two EV fractions from a single trapping run: acoustically trapped EVs and EVs bound specifically to anti-CD9 silica beads. We benchmark our technique against a manual (90 min) immunoaffinity isolation with multiple centrifugation washes. Nanoparticle tracking analysis and transmission electron microscopy with CD9<sup>+</sup> immunogold labeling confirmed isolation of intact EVs. Quantitative proteomic profiling showed that immuno-acoustic fractions contained proteins more strongly associated with CD9 than both acoustically and immunoaffinity-isolated fractions. This methodology can be adapted to isolate specific EV subpopulations expressing other surface proteins while simultaneously collecting a broad EV population as background reference.</p>}},
  author       = {{Broman, Axel and Havers, Megan and Sattarov, Roman and Laurell, Thomas}},
  issn         = {{2045-2322}},
  keywords     = {{Acoustic trapping; Extracellular vesicles; Immuno-acoustic trapping; Immunoaffinity; Proteomics; Subpopulation}},
  language     = {{eng}},
  number       = {{1}},
  publisher    = {{Nature Publishing Group}},
  series       = {{Scientific Reports}},
  title        = {{Immuno-acoustic trapping for extracellular vesicle subpopulations}},
  url          = {{http://dx.doi.org/10.1038/s41598-025-33842-6}},
  doi          = {{10.1038/s41598-025-33842-6}},
  volume       = {{15}},
  year         = {{2025}},
}