D. Božič, K. Vrabec, A. Železnik, A. Raspor, V. Novak, I. Petrović Koshmak, M. Leskovec, and A. Štrancar
International Journal of Molecular Sciences, 2025, Volume 26, Issue 17, Article 8477
Abstract:
The article provides a framework of multiparametric analytical method development for characterization of extracellular vesicle (EV) preparations using size-exclusion chromatography (SEC). The study integrates absorbance, fluorescence, and light scattering detection with SEC to extract information on EV particle content, size, protein content, and surface markers (CD9, CD63, CD81).
A key component of this approach is PATfix® Chromatography System (Sartorius BIA Separations, Slovenia), a chromatography platform equipped with UV, fluorescence, conductivity/pH, and multi-angle light scattering (MALS) detectors, which enabled simultaneous monitoring of these parameters.
The article concludes that the SEC method, enhanced by the PATfix® System, offers a robust protocol for EV characterization. It highlights the system’s potential to improve quality control and reproducibility in EV research, production monitoring, and therapeutic applications. The study also evaluates critical factors for assay robustness, such as mobile phase composition, immunolabelling parameters, and sample preparation conditions, underscoring the system’s role in optimizing these aspects.
