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Article Dans Une Revue Journal of Chromatography A Année : 2023

Fast, simple and calibration-free size characterization and quality control of extracellular vesicles using capillary Taylor dispersion analysis

Sameh Obeid
Thanh Duc Mai
Melissa Reyre
  • Fonction : Auteur
Zuzana Krupova
  • Fonction : Auteur
Pierre Defrenaix
  • Fonction : Auteur
Myriam Taverna

Résumé

This study reports the development of a Taylor Dispersion Analysis (TDA) method for the size characterization of Extracellular Vesicles (EVs), which are highly heterogeneous nanoscale cellderived vesicles (30-1000 nm). Here, we showed that TDA, conducted in uncoated fused silica capillary (50 µm i.d.) using a conventional Capillary Electrophoresis instrument, is able to provide absolute sizing (requiring no calibration) of bovine milk-derived EVs in a small sample volume (~ 7 nL) and over their entire size range, even the smallest ones (< 70 nm) not accessible via other techniques that provide nanoparticle sizing in suspension. TDA size measurements were repeatable (RSD < 10%) and the average EV sizes were found in the range of 120-210 nm, in very good agreement with those measured with Nanoparticle Tracking Analysis, commonly used for EV characterization. TDA allowed quantitative estimation of EVs for concentrations ≥ 2x10 11 EVs/mL. Furthermore, TDA was able to detect minor changes in EV size (i.e. by ~25 nm upon interaction with specific anti-CD9 antibodies of ~150 kDa), and to highlight the impact of extraction methods (i.e. milk pretreatment: freezing, acid precipitation or centrifugation; the type of size-exclusion chromatography column) and of fluorescent labeling (i.e. intravesicular or surface labeling) on the isolated EV population size. In parallel to EV sizing, TDA allowed to detect molecular contaminants (average sizes ~1-13 nm) present within the sample, rendering this method a valuable tool to assess the quality and quantity of EV isolates.
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Dates et versions

hal-04176156 , version 1 (02-08-2023)

Identifiants

Citer

Sameh Obeid, Joseph Chamieh, Thanh Duc Mai, Marco Morani, Melissa Reyre, et al.. Fast, simple and calibration-free size characterization and quality control of extracellular vesicles using capillary Taylor dispersion analysis. Journal of Chromatography A, 2023, 1705, pp.464189. ⟨10.1016/j.chroma.2023.464189⟩. ⟨hal-04176156⟩
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