Affinity-Based Isolation and One-Pot Analysis of Extracellular Vesicles from Biofluids Using Phase Separated...

Area of Science:

• Biotechnology
• Materials Science
• Biochemistry

Background:

• Extracellular vesicles (EVs) are crucial for diagnostics and therapeutics.
• Isolating EVs from complex biofluids like urine is challenging due to their heterogeneity.
• Current isolation methods struggle with efficiency, selectivity, and removing contaminants like lipoproteins.

Purpose of the Study:

• To develop a novel method for efficient and selective isolation of EVs.
• To demonstrate the utility of functionalized zwitterionic (ZW) coacervates for EV isolation and analysis.
• To overcome limitations of conventional EV separation techniques.

Main Methods:

• Development of phase-separated zwitterionic (ZW) coacervates.
• Functionalization of ZW coacervates with a membrane-sensing peptide for affinity-based EV capture.
• Application of the method to isolate EVs from urine samples.
• Analysis of isolated EVs using flow cytometry.

Main Results:

• ZW coacervates provide selective EV isolation across various pH and ionic strengths.
• The method achieves high yields of structurally intact EVs from urine.
• Efficient separation of EVs from lipoproteins was demonstrated.
• A one-pot assay for EV biomarker profiling was successfully implemented, streamlining workflows.

Conclusions:

• Functionalized ZW coacervates offer an effective strategy for selective EV isolation and direct analysis.
• This approach advances EV-based liquid biopsy and drug delivery applications.
• The method paves the way for improved diagnostics and large-scale EV manufacturing.