From:  Unlocking the potential of circulating small extracellular vesicles in neurodegenerative disease through targeted biomarkers and advancements in biosensing
 

Summary of common sEV isolation techniques

Isolation methodAdvantagesDisadvantages
Ultracentrifugation

  • Well-established

  • Widely used

  • Simple protocol

  • Multi-step process

  • Time-consuming

  • Special equipment required

  • Low yields

  • Isolated sEVs could be contaminated with other EVs

Differential filtration

  • Fast, convenient, and low-cost

  • Doesn’t need special equipment

  • sEVs loss due to the entrapment of the sEVs in the filters

  • Cross-contamination with other EVs

SEC

  • Fast, convenient, and low-cost

  • Yields high purity sample

  • Typically, needs to be combined with another method

Polymer-based precipitation

  • Easy protocol

  • High yield

  • Low-cost method

  • Doesn’t need special equipment

  • Cross-contamination with protein aggregates and other EVs

  • Isolated sEVs could be contaminated with the polymer which might affect sEVs characterization, such as zeta potential, and integrity

Immunoprecipitation

  • High purity

  • Doesn’t need special equipment

  • Based on antigen-antibody interactions, which can discriminate between sEV sub-types and isolate particles of interest

  • Relies on high-cost antibodies

  • Low yields

Microfluidic separation

  • Doesn’t need special equipment once fabricated

  • Fast and cost-effective

  • Low yields

  • Complex designs/fabrication

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