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Scalable Isolation and Purification of Extracellular Vesicles from Escherichia coli and Other Bacteria
Published on: October 13, 2021
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A scalable filtration-based method for isolating exomeres and other nanoscale extracellular particles
Thomas Scarborough1, Yuki Kawai-Harada2,3, Olivia Brennan1
1Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, Michigan.
Biorxiv : the Preprint Server for Biology
|November 24, 2025
Summary
Tangential flow filtration efficiently isolates exomeres, a type of non-vesicular extracellular particle (NVEP) that delivers therapeutic RNAs. Exomere miRNAs show remarkable stability, suggesting potential for advanced RNA therapeutics.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Extracellular particles, including extracellular vesicles (EVs) and non-vesicular extracellular particles (NVEPs), mediate intercellular communication via biomolecule transfer.
- Exomeres, a class of NVEPs, show promise for therapeutic RNA delivery due to efficient miRNA transfer.
- Current limitations in exomere isolation hinder their therapeutic application.
Purpose of the Study:
- To evaluate tangential flow filtration (TFF) as a scalable method for exomere isolation.
- To compare TFF-isolated exomeres with those obtained via ultracentrifugation.
- To characterize the stability and miRNA content of TFF-isolated exomeres.
Main Methods:
- Investigated tangential flow filtration (TFF) for separating extracellular particles based on size.
- Utilized differential ultracentrifugation as a benchmark isolation method.
- Analyzed isolated particle composition, including RNAi proteins and miRNAs, and assessed miRNA stability against nucleases, proteases, and surfactants.
Main Results:
- TFF successfully isolated exomeres, demonstrating enrichment in RNAi components (AGO2, HSP90AB1) and unique miRNAs compared to EVs.
- Exomere miRNAs exhibited significant resistance to nuclease degradation, even after protease and surfactant treatment.
- TFF isolation purity and identity were comparable to differential ultracentrifugation.
Conclusions:
- Tangential flow filtration is an efficient and scalable method for isolating exomeres.
- Exomeres represent stable, non-vesicular complexes with potential for extended in vivo half-lives.
- TFF-based exomere isolation could enable bioprocesses for novel RNA therapeutics, potentially outperforming current standards.
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