Affinity‐Based Isolation and One‐Pot Analysis of Extracellular Vesicles from Biofluids Using Phase Separated Zwitterionic Coacervates
Abstract Extracellular vesicles (EVs) hold promise for liquid biopsy and drug delivery applications. However, their heterogeneous nature poses challenges for efficient and selective isolation from complex biofluids. Here, an isolation method based on phase‐separated zwitterionic (ZW) coacervates is...
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| Format: | Article |
| Language: | English |
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Wiley
2025-05-01
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| Series: | Advanced Science |
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| Online Access: | https://doi.org/10.1002/advs.202411653 |
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| author | Francesca Torrini Roberto Frigerio Jonathan Garlipp Philippe Lenzen Karl Normak Carolina Paganini Marina Cretich Alessandro Gori Paolo Arosio |
| author_facet | Francesca Torrini Roberto Frigerio Jonathan Garlipp Philippe Lenzen Karl Normak Carolina Paganini Marina Cretich Alessandro Gori Paolo Arosio |
| author_sort | Francesca Torrini |
| collection | DOAJ |
| description | Abstract Extracellular vesicles (EVs) hold promise for liquid biopsy and drug delivery applications. However, their heterogeneous nature poses challenges for efficient and selective isolation from complex biofluids. Here, an isolation method based on phase‐separated zwitterionic (ZW) coacervates is developed. These coacervates form over a wide range of pH values and ionic strengths, ensuring compatibility with all biofluids. They exhibit antifouling properties that minimize nonspecific binding, allowing for the selective isolation of EVs from biofluids upon functionalization of the polymer with a suitable affinity probe, as proved here with a membrane‐sensing peptide. This strategy is applied to pull down, concentrate, and release EVs from urine samples with high yields while retaining their structural integrity. This approach effectively separates EVs from lipoproteins, a challenging task for conventional separation techniques. The power of the approach is demonstrated as a preparative step for downstream analysis and as a one‐pot assay to profile EV biomarkers in complex fluids. The latter application, implemented here with flow cytometry, significantly streamlines pre‐analytical workflows. Thus, functionalized ZW coacervates represent an effective strategy for the selective isolation and direct analysis of EVs from complex mixtures, paving the way for advances in large‐scale manufacturing and diagnostics. |
| format | Article |
| id | doaj-art-856d100d15094a8a8844ad8f49a5e5b2 |
| institution | DOAJ |
| issn | 2198-3844 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Wiley |
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| series | Advanced Science |
| spelling | doaj-art-856d100d15094a8a8844ad8f49a5e5b22025-08-20T03:20:10ZengWileyAdvanced Science2198-38442025-05-011220n/an/a10.1002/advs.202411653Affinity‐Based Isolation and One‐Pot Analysis of Extracellular Vesicles from Biofluids Using Phase Separated Zwitterionic CoacervatesFrancesca Torrini0Roberto Frigerio1Jonathan Garlipp2Philippe Lenzen3Karl Normak4Carolina Paganini5Marina Cretich6Alessandro Gori7Paolo Arosio8Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering ETH Zurich Zurich 8093 SwitzerlandConsiglio Nazionale delle Ricerche Istituto di Scienze e Tecnologie Chimiche “Giulio Natta” (SCITEC) Milan 20131 ItalyDepartment of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering ETH Zurich Zurich 8093 SwitzerlandDepartment of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering ETH Zurich Zurich 8093 SwitzerlandDepartment of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering ETH Zurich Zurich 8093 SwitzerlandDepartment of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering ETH Zurich Zurich 8093 SwitzerlandConsiglio Nazionale delle Ricerche Istituto di Scienze e Tecnologie Chimiche “Giulio Natta” (SCITEC) Milan 20131 ItalyConsiglio Nazionale delle Ricerche Istituto di Scienze e Tecnologie Chimiche “Giulio Natta” (SCITEC) Milan 20131 ItalyDepartment of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering ETH Zurich Zurich 8093 SwitzerlandAbstract Extracellular vesicles (EVs) hold promise for liquid biopsy and drug delivery applications. However, their heterogeneous nature poses challenges for efficient and selective isolation from complex biofluids. Here, an isolation method based on phase‐separated zwitterionic (ZW) coacervates is developed. These coacervates form over a wide range of pH values and ionic strengths, ensuring compatibility with all biofluids. They exhibit antifouling properties that minimize nonspecific binding, allowing for the selective isolation of EVs from biofluids upon functionalization of the polymer with a suitable affinity probe, as proved here with a membrane‐sensing peptide. This strategy is applied to pull down, concentrate, and release EVs from urine samples with high yields while retaining their structural integrity. This approach effectively separates EVs from lipoproteins, a challenging task for conventional separation techniques. The power of the approach is demonstrated as a preparative step for downstream analysis and as a one‐pot assay to profile EV biomarkers in complex fluids. The latter application, implemented here with flow cytometry, significantly streamlines pre‐analytical workflows. Thus, functionalized ZW coacervates represent an effective strategy for the selective isolation and direct analysis of EVs from complex mixtures, paving the way for advances in large‐scale manufacturing and diagnostics.https://doi.org/10.1002/advs.202411653extracellular vesiclesisolationseparationliquid biopsyzwitterionic coacervatesbiofluids |
| spellingShingle | Francesca Torrini Roberto Frigerio Jonathan Garlipp Philippe Lenzen Karl Normak Carolina Paganini Marina Cretich Alessandro Gori Paolo Arosio Affinity‐Based Isolation and One‐Pot Analysis of Extracellular Vesicles from Biofluids Using Phase Separated Zwitterionic Coacervates Advanced Science extracellular vesicles isolation separation liquid biopsy zwitterionic coacervates biofluids |
| title | Affinity‐Based Isolation and One‐Pot Analysis of Extracellular Vesicles from Biofluids Using Phase Separated Zwitterionic Coacervates |
| title_full | Affinity‐Based Isolation and One‐Pot Analysis of Extracellular Vesicles from Biofluids Using Phase Separated Zwitterionic Coacervates |
| title_fullStr | Affinity‐Based Isolation and One‐Pot Analysis of Extracellular Vesicles from Biofluids Using Phase Separated Zwitterionic Coacervates |
| title_full_unstemmed | Affinity‐Based Isolation and One‐Pot Analysis of Extracellular Vesicles from Biofluids Using Phase Separated Zwitterionic Coacervates |
| title_short | Affinity‐Based Isolation and One‐Pot Analysis of Extracellular Vesicles from Biofluids Using Phase Separated Zwitterionic Coacervates |
| title_sort | affinity based isolation and one pot analysis of extracellular vesicles from biofluids using phase separated zwitterionic coacervates |
| topic | extracellular vesicles isolation separation liquid biopsy zwitterionic coacervates biofluids |
| url | https://doi.org/10.1002/advs.202411653 |
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