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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Materials Science

Background:

  • Cell membrane vesicles (CMVs) are promising drug delivery vehicles, typically produced by extrusion.
  • Extrusion methods result in random vesicle orientations (right-side-out and inside-out).
  • Lack of methods to orient or sort coreless CMVs hinders their therapeutic potential.

Purpose of the Study:

  • To develop a method for orienting and sorting coreless CMVs.
  • To demonstrate the utility of DNA probes for vesicle orientation control.
  • To identify efficient sorting techniques for right-side-out vesicles (RSVs).

Main Methods:

  • Utilized functional DNA probes to detect and sort CMVs based on membrane orientation.
  • Investigated strand displacement reaction, photo cleavage (PC), and enzymatic cleavage for RSV enrichment.
  • Quantified enrichment efficiency and purity of RSVs using different sorting methods.

Main Results:

  • Confirmed random distribution of membrane orientations in extruded coreless CMVs.
  • DNA probes successfully acted as handles for sorting RSVs from inside-out vesicles (ISVs).
  • Photo cleavage (PC) achieved the highest enrichment efficiency (~93%) and RSV purity (~85.4%).

Conclusions:

  • Demonstrated a novel technical platform for orienting and sorting coreless CMVs.
  • PC method is recommended for efficient enrichment of desired vesicle orientations.
  • This work advances CMV technology for diverse biomedical applications.