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Rapid Characterization and Quantification of Extracellular Vesicles by Fluorescence‐Based Microfluidic Diffusion Sizing
Author(s) -
Paganini Carolina,
Hettich Britta,
Kopp Marie R.G.,
Eördögh Adam,
Capasso Palmiero Umberto,
Adamo Giorgia,
Touzet Nicolas,
Manno Mauro,
Bongiovanni Antonella,
RiveraFuentes Pablo,
Leroux JeanChristophe,
Arosio Paolo
Publication year - 2022
Publication title -
advanced healthcare materials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.288
H-Index - 90
eISSN - 2192-2659
pISSN - 2192-2640
DOI - 10.1002/adhm.202100021
Subject(s) - microfluidics , extracellular vesicles , bioprocess , characterization (materials science) , materials science , nanotechnology , biological system , diffusion , reproducibility , sizing , biomolecule , chromatography , chemistry , chemical engineering , biology , physics , organic chemistry , engineering , microbiology and biotechnology , thermodynamics
Extracellular vesicles (EVs) are emerging as promising diagnostic and therapeutic tools for a variety of diseases. The characterization of EVs requires a series of orthogonal techniques that are overall time‐ and material‐consuming. Here, a microfluidic device is presented that exploits the combination of diffusion sizing and multiwavelength fluorescence detection to simultaneously provide information on EV size, concentration, and composition. The latter is achieved with the nonspecific staining of lipids and proteins combined with the specific staining of EV markers such as EV‐associated tetraspanins via antibodies. The device can be operated as a single‐step immunoassay thanks to the integrated separation and quantification of free and EV‐bound fluorophores. This microfluidic technique is capable of detecting and quantifying components associated to EV subtypes and impurities and thus to measure EV purity in a time scale of minutes, requiring less than 5 µL of sample and minimal sample handling before the analysis. Moreover, the analysis is performed directly in solution without immobilization steps. Therefore, this method can accelerate screening of EV samples and aid the evaluation of sample reproducibility, representing an important complementary tool to the current array of biophysical methods for EV characterization, particularly valuable for instance for bioprocess development.