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There is an urgent need to develop techniques for isolation to appraise extracellular vesicles (EVs) in diagnostics and therapeutics. One major challenge in the EV field is to improve and standardize methods of EV isolation and characterization to obtain pure and intact EVs on a large scale. Commonly used EV methods, such as ultracentrifugation, size exclusion chromatography, ultrafiltration, immunoaffinity, precipitation, and microfluidics, each have their advantages and limitations. Here, we present a tangential flow filtration (TFF)-based, multistep purification protocol to isolate EVs at high purity from large volumes of cell culture media. Clinical importance necessitates collecting the maximum amount of EVs, and to achieve the diagnostic goal, EVs should be isolated using techniques that provide maximum yield to confirm that the analyzed results are not misleading.

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Large-Scale Isolation of Extracellular Vesicles Using Tangential Flow Filtration Method

  • 实验方案

  • 发表日期：

    2026年

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  • Mandeep Kaur,
  • Sowmya Shree Gopal,
  • Zenia Barretto,
  • Amit K. Srivastava

摘要

There is an urgent need to develop techniques for isolation to appraise extracellular vesicles (EVs) in diagnostics and therapeutics. One major challenge in the EV field is to improve and standardize methods of EV isolation and characterization to obtain pure and intact EVs on a large scale. Commonly used EV methods, such as ultracentrifugation, size exclusion chromatography, ultrafiltration, immunoaffinity, precipitation, and microfluidics, each have their advantages and limitations. Here, we present a tangential flow filtration (TFF)-based, multistep purification protocol to isolate EVs at high purity from large volumes of cell culture media. Clinical importance necessitates collecting the maximum amount of EVs, and to achieve the diagnostic goal, EVs should be isolated using techniques that provide maximum yield to confirm that the analyzed results are not misleading.