Rapid and efficient isolation platform for plasma extracellular vesicles: EV-FISHER

Area of Science:

• Biotechnology
• Nanotechnology
• Theranostics

Background:

• Extracellular vesicles (EVs) are crucial for clinical theranostics but current isolation methods from plasma are inefficient and time-consuming.
• Existing techniques often involve complex procedures and yield limited amounts of EVs.

Purpose of the Study:

• To develop a rapid, efficient, and user-friendly platform for isolating plasma EVs.
• To demonstrate the platform's superiority over traditional ultracentrifugation methods.
• To validate the platform's utility in downstream applications like single-EV flow cytometry for cancer diagnostics.

Main Methods:

• Development of EV-FISHER, a platform utilizing metal-organic frameworks with cleavable lipid probes (PSDC).
• EV capture via cholesterol interaction and separation using standard centrifugation.
• EV release through enzymatic cleavage of the PSDC probes by deoxyribonuclease I.

Main Results:

• EV-FISHER isolation time is significantly reduced (∼40 min vs. 240 min for UC).
• Isolation efficiency is markedly improved (74.2% vs. 18.1% for UC).
• Lower centrifugation force required (12,800 g vs. 135,000 g for UC).
• Demonstrated compatibility with single-EV flow cytometry for identifying GPC-1 EVs.
• Successful application in breast cancer cohorts for diagnosis and monitoring.

Conclusions:

• EV-FISHER offers a superior alternative to ultracentrifugation for plasma EV isolation.
• The platform enables efficient EV isolation from complex biological fluids.
• This advancement holds significant potential for facilitating EV-based theranostics and diagnostics.