Rapid Characterization and Quantification of Extracellular Vesicles by Fluorescence‐Based Microfluidic Diffusion Sizing
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Date
2022-03-02
Publication Type
Journal Article
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yes
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Abstract
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.
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published
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Journal / series
Volume
11 (5)
Pages / Article No.
2100021
Publisher
Wiley-VCH
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Methods
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Date collected
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Subject
characterization; extracellular vesicles; fluorescence; immunoassay; microfluidics; purity; quantification
Organisational unit
09572 - Arosio, Paolo / Arosio, Paolo
03811 - Leroux, Jean-Christophe / Leroux, Jean-Christophe
Notes
Funding
801338 - Extracellular vesicles from a natural source for tailor-made nanomaterials (EC)
ETH-33 17-2 - Microfluidic assays for the quantification of low concentrations of proteins in heterogeneous mixtures (ETHZ)
- Intradermal delivery of stem cell exosomes ()
ETH-33 17-2 - Microfluidic assays for the quantification of low concentrations of proteins in heterogeneous mixtures (ETHZ)
- Intradermal delivery of stem cell exosomes ()