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Microfluidic device for high-throughput affinity-based isolation of extracellular vesicles.

Ting-Wen Lo1, Ziwen Zhu, Emma Purcell

  • 1Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, USA. snagrath@umich.edu.

Lab on a Chip
|April 28, 2020
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Summary

Researchers developed a high-throughput method for isolating extracellular vesicles (EVs) using a modified microfluidic chip. This new technique allows for the efficient capture and release of functional EVs, overcoming limitations of current immunoaffinity methods.

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Cell Biology

Background:

  • Immunoaffinity isolation offers high specificity for extracellular vesicles (EVs) but faces challenges in EV release and throughput.
  • Current methods often require harsh conditions for EV elution, compromising their function, and suffer from low flow rates, limiting sample processing.
  • Efficient and gentle isolation of intact EVs is crucial for downstream biological studies and biomarker discovery.

Purpose of the Study:

  • To develop a high-throughput and gentle method for isolating extracellular vesicles (EVs) using immunoaffinity capture.
  • To enable the release of captured EVs in a functional state for subsequent analyses.
  • To adapt a microfluidic device originally designed for circulating tumor cell isolation for efficient EV isolation.

Main Methods:

  • Modification of the OncoBean chip, a radial flow microfluidic device, by functionalizing bean-shaped microposts with antibodies targeting common EV surface markers.
  • Utilizing desthiobiotin-conjugated antibodies to facilitate the release of captured EVs under mild conditions.
  • Assessing the functionality of released EVs through cellular uptake studies using flow cytometry and fluorescent microscopy.

Main Results:

  • The modified OncoBean chip achieved high-throughput isolation of EVs by leveraging its unique design for increased surface area and varying shear rates.
  • The incorporation of desthiobiotin-conjugated antibodies enabled the successful release of captured EVs, a significant improvement over existing methods.
  • Released EVs demonstrated functionality, confirmed by their uptake into cells, indicating preserved biological activity.

Conclusions:

  • The developed immunoaffinity-based, high-throughput EV isolation technology overcomes critical limitations of existing methods, particularly regarding EV release and processing speed.
  • This approach facilitates the study of EVs as crucial mediators of intercellular communication.
  • The technology holds promise for advancing the identification and characterization of EV biomarkers for diagnostic and therapeutic applications.