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Updated: Jul 31, 2025

Purification of High Yield Extracellular Vesicle Preparations Away from Virus

Purification of High Yield Extracellular Vesicle Preparations Away from Virus

Published on: September 12, 2019

Extracellular Vesicle Isolation by a Tangential-Flow Filtration-Based Large-Scale Purification Method.

Runjie Yuan¹, Yijun Zhou¹, Gabriel F Arias²

¹Department of Microbiology and Immunology, The University of North Carolina, Chapel Hill, NC, USA.

Methods in Molecular Biology (Clifton, N.J.)

|May 4, 2023

Summary

Isolating pure extracellular vesicles (EVs) at scale is challenging. This study introduces a multi-step protocol using tangential-flow filtration (TFF) and multimodal chromatography (MMC) for high-purity EV isolation from large volumes.

Keywords:

Capto Core 700 multimodal chromatography (MMC)Extracellular vesicles (EVs)Purification Tangential-flow filtration (TFF)

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Last Updated: Jul 31, 2025

Purification of High Yield Extracellular Vesicle Preparations Away from Virus

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Published on: September 12, 2019

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Scalable Isolation and Purification of Extracellular Vesicles from Escherichia coli and Other Bacteria

Scalable Isolation and Purification of Extracellular Vesicles from Escherichia coli and Other Bacteria

Published on: October 13, 2021

Area of Science:

Biotechnology
Cell Biology
Biochemistry

Background:

Extracellular vesicle (EV) isolation from cell culture media is difficult, especially at scale.
Existing methods like ultracentrifugation and PEG precipitation have limitations in purity and yield.
Obtaining pure, intact EVs is crucial for research and therapeutic applications.

Purpose of the Study:

To develop an efficient, scalable, and high-purity protocol for isolating extracellular vesicles (EVs).
To overcome the limitations of conventional EV isolation techniques.
To present a multi-step purification strategy combining tangential-flow filtration (TFF), polyethylene glycol (PEG) precipitation, and multimodal chromatography (MMC).

Main Methods:

A multi-step purification protocol was developed.
Tangential-flow filtration (TFF) was employed to remove soluble proteins before precipitation.
Polyethylene glycol (PEG) precipitation was combined with Capto Core 700 multimodal chromatography (MMC) for EV purification.

Main Results:

The TFF step effectively removed proteins, preventing aggregation and co-purification.
The combined method achieved high purity of isolated EVs.
The protocol is suitable for large-scale EV isolation from conditioned cell culture medium.

Conclusions:

The presented TFF-based multi-step protocol offers a robust method for high-purity EV isolation.
This approach addresses the challenges of scalability and purity in EV purification.
The method enhances the quality of EVs for downstream applications.