ArticlePLoS neglected tropical diseases2019
The importance of extracellular vesicle purification for downstream analysis: A comparison of differential centrifugation and size exclusion chromatography for helminth pathogens.
Article in PLoS neglected tropical diseases, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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Who cites it
40 citing papers in PubMed, 76 citations in OpenAlex.
- Thyroid-Originating Extracellular Vesicles Harbor Thyroid-Specific Biomarkers with Potential Relevance for Thyroid Cancer Recurrence Detection.International journal of molecular sciences · 2026Article
- Plant-Derived Exosomes: Nano-Inducers of Cross-Kingdom Regulations.Pharmaceuticals (Basel, Switzerland) · 2025Review
- Detoxome Capacity of the Adult Rumen FlukeJournal of proteome research · 2025Article
- Establishment of minimum protein standards forMycobacteria (London, England) · 2025Article
- Proteomic and metabolomic profiles of plasma-derived Extracellular Vesicles differentiate melanoma patients from healthy controls.Translational oncology · 2024Article
- Characterisation of extracellular vesicles in baculovirus infection ofJournal of extracellular biology · 2024Article
- Article
- A Comparative Analysis of the Protein Cargo of Extracellular Vesicles from Helminth Parasites.Life (Basel, Switzerland) · 2023Article
- Guidelines for the purification and characterization of extracellular vesicles of parasites.Journal of extracellular biology · 2023Article
- Emerging Roles of Using Small Extracellular Vesicles as an Anti-Cancer Drug.International journal of molecular sciences · 2023Review
- Assessment of anti-inflammatory bioactivity of extracellular vesicles is susceptible to error via media component contamination.Cytotherapy · 2023Article
- Therapeutic Implications of the Drug Resistance Conferred by Extracellular Vesicles Derived from Triple-Negative Breast Cancer Cells.International journal of molecular sciences · 2023Review
- Proteomic analysis of exosome-like vesicles from Fasciola gigantica adult worm provides support for new vaccine targets against fascioliasis.Parasites & vectors · 2023Article
- Analyzing bronchoalveolar fluid derived small extracellular vesicles using single-vesicle SERS for non-small cell lung cancer detection.Sensors & diagnostics · 2023Article
- Special considerations for studies of extracellular vesicles from parasitic helminths: A community-led roadmap to increase rigour and reproducibility.Journal of extracellular vesicles · 2023Article
- Circulating Exosomes from Alzheimer's Disease Suppress Vascular Endothelial-Cadherin Expression and Induce Barrier Dysfunction in Recipient Brain Microvascular Endothelial Cell.Journal of Alzheimer's disease : JAD · 2023Article
- Excretory-secretory products from the brown stomach worm, Teladorsagia circumcincta, exert antimicrobial activity in in vitro growth assays.Parasites & vectors · 2022Article
- Article
- Extending applications of AFM to fluidic AFM in single living cell studies.Journal of cellular physiology · 2022Review
- Macrophage-Derived Extracellular Vesicles: A Promising Tool for Personalized Cancer Therapy.Biomedicines · 2022Review
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
backgroundRobust protocols for the isolation of extracellular vesicles (EVs) from the rest of their excretory-secretory products are necessary for downstream studies and application development. The most widely used purification method of EVs for helminth pathogens is currently differential centrifugation (DC). In contrast, size exclusion chromatography (SEC) has been included in the purification pipeline for EVs from other pathogens, highlighting there is not an agreed research community 'gold standard' for EV isolation. In this case study, Fasciola hepatica from natural populations were cultured in order to collect EVs from culture media and evaluate a SEC or DC approach to pathogen helminth EV purification. METHODOLOGY/PRINCIPAL
findingsTransmission electron and atomic force microscopy demonstrated that EVs prepared by SEC were both smaller in size and less diverse than EV resolved by DC. Protein quantification and Western blotting further demonstrated that SEC purification realised a higher EV purity to free excretory-secretory protein (ESP) yield ratio compared to DC approaches as evident by the reduction of soluble free cathepsin L proteases in SEC EV preparations. Proteomic analysis further highlighted DC contamination from ESP as shown by an increased diversity of protein identifications and unique peptide hits in DC EVs as compared to SEC EVs. In addition, SEC purified EVs contained less tegumental based proteins than DC purified EVs. CONCLUSIONS/SIGNIFICANCE: The data suggests that DC and SEC purification methods do not isolate equivalent EV population profiles and caution should be taken in the choice of EV purification utilised, with certain protocols for DC preparations including more free ES proteins and tegumental artefacts. We propose that SEC methods should be used for EV purification prior to downstream studies.
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