Multi-parametric orthogonal characterization of extracellular vesicles by liquid chromatography combined with in-line light scattering and fluorescence detection

10 May 2023, Version 1
This content is a preprint and has not undergone peer review at the time of posting.


Extracellular vesicles (EVs) are membrane enclosed biological nanoparticles with potential as diagnostic markers and carriers for therapeutics. Characterization of EVs poses severe challenges due to their complex structure and composition, requiring the combination of orthogonal analytical techniques. Here, we demonstrate how liquid chromatography combined with multi-angle light scattering (MALS) and fluorescence detection can provide multi-parametric characterization of EV samples, including concentration of particles, average diameter of the particles, protein amount to particle number ratio, presence of EV surface markers and lipids, EV shape and sample purity. The method requires a small amount of sample of approximately 107 EVs, limited handling of the sample and data analysis time in the order of minutes; it is fully automatable and can be applied to both crude and purified samples.


Extracellular vesicles
Multi angle light scattering
Size exclusion chromatography
Protein quantification
Particle quantification
Particle sizing
Anion exchange chromatography

Supplementary materials

Supplementary Information
Supplementary information accompanying the main text. It includes the derivation of the equations used in the main work as well as additional figures referred to in the main text.


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