Skip to main content
Research article - Peer-reviewed, 2022

Coincident Fluorescence-Burst Analysis of the Loading Yields of Exosome-Mimetic Nanovesicles with Fluorescently-Labeled Cargo Molecules

Sanaee, Maryam; Sandberg, Elin; Ronquist, K. Goran; Morrell, Jane M.; Widengren, Jerker; Gallo, Katia

Abstract

The possible targeting functionality and low immunogenicity of exosomes and exosome-like nanovesicles make them promising as drug-delivery carriers. To tap into this potential, accurate non-destructive methods to load them and characterize their contents are of utmost importance. However, the small size, polydispersity, and aggregation of nanovesicles in solution make quantitative characterizations of their loading particularly challenging. Here, an ad-hoc methodology is developed based on burst analysis of dual-color confocal fluorescence microscopy experiments, suited for quantitative characterizations of exosome-like nanovesicles and of their fluorescently-labeled loading. It is applied to study exosome-mimetic nanovesicles derived from animal extracellular-vesicles and human red blood cell detergent-resistant membranes, loaded with fluorescently-tagged dUTP cargo molecules. For both classes of nanovesicles, successful loading is proved and by dual-color coincident fluorescence burst analysis, size statistics and loading yields are retrieved and quantified. The procedure affords single-vesicle characterizations well-suited for the investigation of a variety of cargo molecules and biological nanovesicle combinations besides the proof-of-principle demonstrations of this study. The results highlight a powerful characterization tool essential for optimizing the loading process and for advanced engineering of biomimetic nanovesicles for therapeutic drug delivery.

Keywords

burst analysis; drug loading; extracellular nanovesicles; fluorescence correlation spectroscopy; single vesicle profiling

Published in

Small
2022, volume: 18, number: 12, article number: 2106241
Publisher: WILEY-V C H VERLAG GMBH

Authors' information

Sanaee, Maryam
Royal Institute of Technology
Sandberg, Elin
Royal Institute of Technology
Swedish University of Agricultural Sciences, Department of Clinical Sciences
Swedish University of Agricultural Sciences, Department of Clinical Sciences
Widengren, Jerker
Royal Institute of Technology
Gallo, Katia
Royal Institute of Technology

UKÄ Subject classification

Pharmaceutical Sciences

Publication Identifiers

DOI: https://doi.org/10.1002/smll.202106241

URI (permanent link to this page)

https://res.slu.se/id/publ/116094