Snorkel‐tag based affinity chromatography for recombinant extracellular vesicle purification
Abstract Extracellular vesicles (EVs) are lipid nanoparticles and play an important role in cell‐cell communications, making them potential therapeutic agents and allowing to engineer for targeted drug delivery. The expanding applications of EVs in next generation medicine is still limited by existi...
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| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2024-10-01
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| Series: | Journal of Extracellular Vesicles |
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| Online Access: | https://doi.org/10.1002/jev2.12523 |
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| _version_ | 1846118091993382912 |
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| author | Madhusudhan Reddy Bobbili André Görgens Yan Yan Stefan Vogt Dhanu Gupta Giulia Corso Samir Barbaria Carolina Patrioli Sylvia Weilner Marianne Pultar Jaroslaw Jacak Matthias Hackl Markus Schosserer Regina Grillari Jørgen Kjems Samir EL Andaloussi Johannes Grillari |
| author_facet | Madhusudhan Reddy Bobbili André Görgens Yan Yan Stefan Vogt Dhanu Gupta Giulia Corso Samir Barbaria Carolina Patrioli Sylvia Weilner Marianne Pultar Jaroslaw Jacak Matthias Hackl Markus Schosserer Regina Grillari Jørgen Kjems Samir EL Andaloussi Johannes Grillari |
| author_sort | Madhusudhan Reddy Bobbili |
| collection | DOAJ |
| description | Abstract Extracellular vesicles (EVs) are lipid nanoparticles and play an important role in cell‐cell communications, making them potential therapeutic agents and allowing to engineer for targeted drug delivery. The expanding applications of EVs in next generation medicine is still limited by existing tools for scaling standardized EV production, single EV tracing and analytics, and thus provide only a snapshot of tissue‐specific EV cargo information. Here, we present the Snorkel‐tag, for which we have genetically fused the EV surface marker protein CD81, to a series of tags with an additional transmembrane domain to be displayed on the EV surface, resembling a snorkel. This system enables the affinity purification of EVs from complex matrices in a non‐destructive form while maintaining EV characteristics in terms of surface protein profiles, associated miRNA patterns and uptake into a model cell line. Therefore, we consider the Snorkel‐tag to be a widely applicable tool in EV research, allowing for efficient preparation of EV standards and reference materials, or dissecting EVs with different surface markers when fusing to other tetraspanins in vitro or in vivo. |
| format | Article |
| id | doaj-art-f0ad57c4757644158df4812e317e786e |
| institution | Kabale University |
| issn | 2001-3078 |
| language | English |
| publishDate | 2024-10-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Vesicles |
| spelling | doaj-art-f0ad57c4757644158df4812e317e786e2024-12-18T07:26:41ZengWileyJournal of Extracellular Vesicles2001-30782024-10-011310n/an/a10.1002/jev2.12523Snorkel‐tag based affinity chromatography for recombinant extracellular vesicle purificationMadhusudhan Reddy Bobbili0André Görgens1Yan Yan2Stefan Vogt3Dhanu Gupta4Giulia Corso5Samir Barbaria6Carolina Patrioli7Sylvia Weilner8Marianne Pultar9Jaroslaw Jacak10Matthias Hackl11Markus Schosserer12Regina Grillari13Jørgen Kjems14Samir EL Andaloussi15Johannes Grillari16Institute of Molecular Biotechnology, Department of Biotechnology BOKU University Vienna AustriaDepartment of Laboratory Medicine, Division of Biomolecular and Cellular Medicine Karolinska Institutet Stockholm SwedenDepartment of Molecular Biology and Genetics, Centre for Cellular Signal Patterns (CellPat), Interdisciplinary Nanoscience Centre (iNANO) Aarhus University Aarhus C DenmarkInstitute of Molecular Biotechnology, Department of Biotechnology BOKU University Vienna AustriaDepartment of Laboratory Medicine, Division of Biomolecular and Cellular Medicine Karolinska Institutet Stockholm SwedenDepartment of Laboratory Medicine, Division of Biomolecular and Cellular Medicine Karolinska Institutet Stockholm SwedenInstitute of Molecular Biotechnology, Department of Biotechnology BOKU University Vienna AustriaInstitute of Molecular Biotechnology, Department of Biotechnology BOKU University Vienna AustriaInstitute of Molecular Biotechnology, Department of Biotechnology BOKU University Vienna AustriaTAmiRNA TAmiRNA GmbH Vienna AustriaLudwig Boltzmann Institute for Traumatology The Research Center in Cooperation with AUVA Vienna AustriaAustrian Cluster for Tissue RegenerationInstitute of Molecular Biotechnology, Department of Biotechnology BOKU University Vienna AustriaAustrian Cluster for Tissue RegenerationDepartment of Molecular Biology and Genetics, Centre for Cellular Signal Patterns (CellPat), Interdisciplinary Nanoscience Centre (iNANO) Aarhus University Aarhus C DenmarkDepartment of Laboratory Medicine, Division of Biomolecular and Cellular Medicine Karolinska Institutet Stockholm SwedenInstitute of Molecular Biotechnology, Department of Biotechnology BOKU University Vienna AustriaAbstract Extracellular vesicles (EVs) are lipid nanoparticles and play an important role in cell‐cell communications, making them potential therapeutic agents and allowing to engineer for targeted drug delivery. The expanding applications of EVs in next generation medicine is still limited by existing tools for scaling standardized EV production, single EV tracing and analytics, and thus provide only a snapshot of tissue‐specific EV cargo information. Here, we present the Snorkel‐tag, for which we have genetically fused the EV surface marker protein CD81, to a series of tags with an additional transmembrane domain to be displayed on the EV surface, resembling a snorkel. This system enables the affinity purification of EVs from complex matrices in a non‐destructive form while maintaining EV characteristics in terms of surface protein profiles, associated miRNA patterns and uptake into a model cell line. Therefore, we consider the Snorkel‐tag to be a widely applicable tool in EV research, allowing for efficient preparation of EV standards and reference materials, or dissecting EVs with different surface markers when fusing to other tetraspanins in vitro or in vivo.https://doi.org/10.1002/jev2.12523affinity chromatographyCD81extracellular vesiclessnorkel‐tagStEVACtetraspanins |
| spellingShingle | Madhusudhan Reddy Bobbili André Görgens Yan Yan Stefan Vogt Dhanu Gupta Giulia Corso Samir Barbaria Carolina Patrioli Sylvia Weilner Marianne Pultar Jaroslaw Jacak Matthias Hackl Markus Schosserer Regina Grillari Jørgen Kjems Samir EL Andaloussi Johannes Grillari Snorkel‐tag based affinity chromatography for recombinant extracellular vesicle purification Journal of Extracellular Vesicles affinity chromatography CD81 extracellular vesicles snorkel‐tag StEVAC tetraspanins |
| title | Snorkel‐tag based affinity chromatography for recombinant extracellular vesicle purification |
| title_full | Snorkel‐tag based affinity chromatography for recombinant extracellular vesicle purification |
| title_fullStr | Snorkel‐tag based affinity chromatography for recombinant extracellular vesicle purification |
| title_full_unstemmed | Snorkel‐tag based affinity chromatography for recombinant extracellular vesicle purification |
| title_short | Snorkel‐tag based affinity chromatography for recombinant extracellular vesicle purification |
| title_sort | snorkel tag based affinity chromatography for recombinant extracellular vesicle purification |
| topic | affinity chromatography CD81 extracellular vesicles snorkel‐tag StEVAC tetraspanins |
| url | https://doi.org/10.1002/jev2.12523 |
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