Methods and Challenges in Purifying Drug‐Loaded Extracellular Vesicles

ABSTRACT Extracellular vesicles (EV) have emerged as promising nanocarriers for drug delivery. However, the efficient loading of therapeutic molecules into EV and the subsequent purification of drug‐loaded EV from unloaded drugs remain significant challenges. This review explores the most used metho...

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Main Authors: Marie Auquière, Giulio G. Muccioli, Anne des Rieux
Format: Article
Language:English
Published: Wiley 2025-06-01
Series:Journal of Extracellular Vesicles
Online Access:https://doi.org/10.1002/jev2.70097
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author Marie Auquière
Giulio G. Muccioli
Anne des Rieux
author_facet Marie Auquière
Giulio G. Muccioli
Anne des Rieux
author_sort Marie Auquière
collection DOAJ
description ABSTRACT Extracellular vesicles (EV) have emerged as promising nanocarriers for drug delivery. However, the efficient loading of therapeutic molecules into EV and the subsequent purification of drug‐loaded EV from unloaded drugs remain significant challenges. This review explores the most used methods for EV purification, meaning the separation of drug‐loaded EV from unloaded drugs, including ultracentrifugation, density gradient centrifugation, ultrafiltration, size exclusion chromatography, dialysis and commercial exosome isolation kits. The principles, advantages and limitations of each method are discussed. Critical parameters such as molecular weight cutoff, membrane composition, and the nature of the loaded molecule are highlighted for their impact on the purification process. The review also addresses the technical aspects, including time, cost and equipment requirements, and emphasizes the need for standardized guidelines to improve reproducibility and comparability across studies. By providing a comprehensive overview of current purification strategies, this review aims to guide researchers in selecting the most appropriate methods for advancing EV‐based drug delivery systems.
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spelling doaj-art-cb0439d087a84110a9a507b05b201eaa2025-08-20T02:48:54ZengWileyJournal of Extracellular Vesicles2001-30782025-06-01146n/an/a10.1002/jev2.70097Methods and Challenges in Purifying Drug‐Loaded Extracellular VesiclesMarie Auquière0Giulio G. Muccioli1Anne des Rieux2Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials (ADDB) Université Catholique de Louvain, UCLouvain Brussels BelgiumLouvain Drug Research Institute Bioanalysis and Pharmacology of Bioactive Lipids (BPBL) Université Catholique de Louvain, UCLouvain Brussels BelgiumLouvain Drug Research Institute, Advanced Drug Delivery and Biomaterials (ADDB) Université Catholique de Louvain, UCLouvain Brussels BelgiumABSTRACT Extracellular vesicles (EV) have emerged as promising nanocarriers for drug delivery. However, the efficient loading of therapeutic molecules into EV and the subsequent purification of drug‐loaded EV from unloaded drugs remain significant challenges. This review explores the most used methods for EV purification, meaning the separation of drug‐loaded EV from unloaded drugs, including ultracentrifugation, density gradient centrifugation, ultrafiltration, size exclusion chromatography, dialysis and commercial exosome isolation kits. The principles, advantages and limitations of each method are discussed. Critical parameters such as molecular weight cutoff, membrane composition, and the nature of the loaded molecule are highlighted for their impact on the purification process. The review also addresses the technical aspects, including time, cost and equipment requirements, and emphasizes the need for standardized guidelines to improve reproducibility and comparability across studies. By providing a comprehensive overview of current purification strategies, this review aims to guide researchers in selecting the most appropriate methods for advancing EV‐based drug delivery systems.https://doi.org/10.1002/jev2.70097
spellingShingle Marie Auquière
Giulio G. Muccioli
Anne des Rieux
Methods and Challenges in Purifying Drug‐Loaded Extracellular Vesicles
Journal of Extracellular Vesicles
title Methods and Challenges in Purifying Drug‐Loaded Extracellular Vesicles
title_full Methods and Challenges in Purifying Drug‐Loaded Extracellular Vesicles
title_fullStr Methods and Challenges in Purifying Drug‐Loaded Extracellular Vesicles
title_full_unstemmed Methods and Challenges in Purifying Drug‐Loaded Extracellular Vesicles
title_short Methods and Challenges in Purifying Drug‐Loaded Extracellular Vesicles
title_sort methods and challenges in purifying drug loaded extracellular vesicles
url https://doi.org/10.1002/jev2.70097
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