Control of Physical and Biochemical Parameters Influencing Exogeneous Cargo Protein Association to Extracellular Vesicles Using Lipid Anchors Enables High Loading and Effective Intracellular Delivery
ABSTRACT Despite biomolecule delivery is a natural function of extracellular vesicles (EVs), low loading of exogenous macromolecules such as proteins into EVs limits their interest as convincing protein delivery systems for health applications. In this context, lipid‐anchorage of exogenous cargo int...
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| Format: | Article |
| Language: | English |
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Wiley
2025-05-01
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| Series: | Journal of Extracellular Biology |
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| Online Access: | https://doi.org/10.1002/jex2.70048 |
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| author | Antonin Marquant Jade Berthelot Claudia Bich Zeineb Ibn Elfekih Laurianne Simon Baptiste Robin Joël Chopineau David Tianpei Wang Samuel Jay Emerson Aijun Wang Clément Benedetti Simon Langlois Laurence Guglielmi Pierre Martineau Anne Aubert‐Pouëssel Marie Morille |
| author_facet | Antonin Marquant Jade Berthelot Claudia Bich Zeineb Ibn Elfekih Laurianne Simon Baptiste Robin Joël Chopineau David Tianpei Wang Samuel Jay Emerson Aijun Wang Clément Benedetti Simon Langlois Laurence Guglielmi Pierre Martineau Anne Aubert‐Pouëssel Marie Morille |
| author_sort | Antonin Marquant |
| collection | DOAJ |
| description | ABSTRACT Despite biomolecule delivery is a natural function of extracellular vesicles (EVs), low loading of exogenous macromolecules such as proteins into EVs limits their interest as convincing protein delivery systems for health applications. In this context, lipid‐anchorage of exogenous cargo into EV membrane recently emerged as a promising option to enable their vectorisation into cells. Nevertheless, this option was not explored for protein intracellular delivery, and further characterisation of critical parameters governing the association of a lipid‐anchored cargo protein to EVs is still needed to confirm the relevance of this anchorage strategy. Therefore, we sought to identify these parameters in a precise and quantitative manner, using bulk and single nanoparticle analysis methods to identify protein loading capacity and subsequent intracellular delivery. We identified incubation temperature, cargo concentration, lipid anchor (LA) structure (lipid nature, linker) and EV origin as critical factors influencing maximal EV loading capacity. Precise control of these parameters enabled to load cargo protein close to EV saturation without hindering cellular delivery. The structural properties of LA influenced not only cargo protein/EV association but also intracellular delivery into different carcinoma cell lines. By thoroughly characterising Lipid‐PEG‐protein anchorage, this study evidences the interest of this tunable and controllable approach for efficient EV protein delivery. |
| format | Article |
| id | doaj-art-604aa66b8ffa4e208cec288f0fcfd3a0 |
| institution | OA Journals |
| issn | 2768-2811 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Biology |
| spelling | doaj-art-604aa66b8ffa4e208cec288f0fcfd3a02025-08-20T01:56:49ZengWileyJournal of Extracellular Biology2768-28112025-05-0145n/an/a10.1002/jex2.70048Control of Physical and Biochemical Parameters Influencing Exogeneous Cargo Protein Association to Extracellular Vesicles Using Lipid Anchors Enables High Loading and Effective Intracellular DeliveryAntonin Marquant0Jade Berthelot1Claudia Bich2Zeineb Ibn Elfekih3Laurianne Simon4Baptiste Robin5Joël Chopineau6David Tianpei Wang7Samuel Jay Emerson8Aijun Wang9Clément Benedetti10Simon Langlois11Laurence Guglielmi12Pierre Martineau13Anne Aubert‐Pouëssel14Marie Morille15ICGM Montpellier University, CNRS, ENSCM Montpellier FranceICGM Montpellier University, CNRS, ENSCM Montpellier FranceInstitut des Biomolécules Max Mousseron (IBMM) Montpellier FranceICGM Montpellier University, CNRS, ENSCM Montpellier FranceICGM Montpellier University, CNRS, ENSCM Montpellier FranceICGM Montpellier University, CNRS, ENSCM Montpellier FranceICGM Montpellier University, CNRS, ENSCM Montpellier FranceCenter for Surgical Bioengineering University of California Davis Sacramento California USACenter for Surgical Bioengineering University of California Davis Sacramento California USACenter for Surgical Bioengineering University of California Davis Sacramento California USAMontpellier Ressources Imagerie, BioCampus University of Montpellier, CNRS, INSERM Montpellier FranceMontpellier Ressources Imagerie, BioCampus University of Montpellier, CNRS, INSERM Montpellier FranceInstitut de Recherche en Cancérologie de Montpellier (IRCM), INSERM Montpellier FranceInstitut de Recherche en Cancérologie de Montpellier (IRCM), INSERM Montpellier FranceICGM Montpellier University, CNRS, ENSCM Montpellier FranceICGM Montpellier University, CNRS, ENSCM Montpellier FranceABSTRACT Despite biomolecule delivery is a natural function of extracellular vesicles (EVs), low loading of exogenous macromolecules such as proteins into EVs limits their interest as convincing protein delivery systems for health applications. In this context, lipid‐anchorage of exogenous cargo into EV membrane recently emerged as a promising option to enable their vectorisation into cells. Nevertheless, this option was not explored for protein intracellular delivery, and further characterisation of critical parameters governing the association of a lipid‐anchored cargo protein to EVs is still needed to confirm the relevance of this anchorage strategy. Therefore, we sought to identify these parameters in a precise and quantitative manner, using bulk and single nanoparticle analysis methods to identify protein loading capacity and subsequent intracellular delivery. We identified incubation temperature, cargo concentration, lipid anchor (LA) structure (lipid nature, linker) and EV origin as critical factors influencing maximal EV loading capacity. Precise control of these parameters enabled to load cargo protein close to EV saturation without hindering cellular delivery. The structural properties of LA influenced not only cargo protein/EV association but also intracellular delivery into different carcinoma cell lines. By thoroughly characterising Lipid‐PEG‐protein anchorage, this study evidences the interest of this tunable and controllable approach for efficient EV protein delivery.https://doi.org/10.1002/jex2.70048cell deliveryEV exogenous engineeringexosomeslipid‐PEG anchorageprocess optimisationprotein association |
| spellingShingle | Antonin Marquant Jade Berthelot Claudia Bich Zeineb Ibn Elfekih Laurianne Simon Baptiste Robin Joël Chopineau David Tianpei Wang Samuel Jay Emerson Aijun Wang Clément Benedetti Simon Langlois Laurence Guglielmi Pierre Martineau Anne Aubert‐Pouëssel Marie Morille Control of Physical and Biochemical Parameters Influencing Exogeneous Cargo Protein Association to Extracellular Vesicles Using Lipid Anchors Enables High Loading and Effective Intracellular Delivery Journal of Extracellular Biology cell delivery EV exogenous engineering exosomes lipid‐PEG anchorage process optimisation protein association |
| title | Control of Physical and Biochemical Parameters Influencing Exogeneous Cargo Protein Association to Extracellular Vesicles Using Lipid Anchors Enables High Loading and Effective Intracellular Delivery |
| title_full | Control of Physical and Biochemical Parameters Influencing Exogeneous Cargo Protein Association to Extracellular Vesicles Using Lipid Anchors Enables High Loading and Effective Intracellular Delivery |
| title_fullStr | Control of Physical and Biochemical Parameters Influencing Exogeneous Cargo Protein Association to Extracellular Vesicles Using Lipid Anchors Enables High Loading and Effective Intracellular Delivery |
| title_full_unstemmed | Control of Physical and Biochemical Parameters Influencing Exogeneous Cargo Protein Association to Extracellular Vesicles Using Lipid Anchors Enables High Loading and Effective Intracellular Delivery |
| title_short | Control of Physical and Biochemical Parameters Influencing Exogeneous Cargo Protein Association to Extracellular Vesicles Using Lipid Anchors Enables High Loading and Effective Intracellular Delivery |
| title_sort | control of physical and biochemical parameters influencing exogeneous cargo protein association to extracellular vesicles using lipid anchors enables high loading and effective intracellular delivery |
| topic | cell delivery EV exogenous engineering exosomes lipid‐PEG anchorage process optimisation protein association |
| url | https://doi.org/10.1002/jex2.70048 |
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