{
  "abstract": "Tangential flow filtration (TFF) is an effective method for scalable EV isolation. Here, we describe and validate a reference protocol for TriPort-TFF, a laboratory-assembled TFF system featuring a three- port reservoir design enabling sample circulation, retentate return, and diafiltration. Constructed from individually sourced, commercially available components with detailed assembly and operating parameters, TriPort-TFF can be built at a lower cost than commercially integrated TFF platforms. We evaluated the system using conditioned media (CM) from human MDM cultured with EV-depleted human serum, human cerebral organoids in serum-free medium, and mouse BMDM cultured with EV-depleted foetal bovine serum (FBS) unstimulated or LPS-stimulated. The system reproducibly isolated EVs from all three sources, and biological replicates of BMDM showed distinct proteomic clusters between LPS-stimulated and unstimulated conditions. The requirement for subsequent size-exclusion chromatography (SEC) after TFF depended on the sample serum-presence. In serum-containing CM, TFF alone co-concentrated serum proteins alongside EVs, subsequent SEC improved the particle-to- protein ratio, reduced inter-replicate variability and improved proteomic depth. TFF alone in serum- free organoid CM already yielded higher particle-to-protein ratios than TFF-only preparations from serum-containing CM, with SEC further improving this ratio. TriPort-TFF offers an accessible system for EV-isolation that can be combined with SEC for diverse sample types.",
  "abstract_shown": true,
  "affiliations": [
    "School of Medical Sciences, University of Sydney, Sydney, NSW, Australia"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 12,
  "authors": [
    {
      "name": "D. Mainali"
    },
    {
      "name": "M. D. Hade"
    },
    {
      "name": "A. Ge"
    },
    {
      "name": "H. Ezzarouli"
    },
    {
      "name": "S. Maksour"
    },
    {
      "name": "L. Thompson"
    },
    {
      "name": "C. L. Moreno"
    },
    {
      "name": "G. G. Neely"
    },
    {
      "name": "S. Hallal"
    },
    {
      "name": "C. Goldsbury"
    },
    {
      "name": "S. Magana"
    },
    {
      "affs": [
        0
      ],
      "name": "L. Piccio"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/f9c5c5pj4j87y.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.10.03.756465",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.10.03.756465v1.full",
  "id": "f9c5c5pj4j87y",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.10.03.756465v1",
  "language": null,
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "cell biology"
    ],
    [
      "openalex",
      "1312|Extracellular vesicles in disease"
    ]
  ],
  "openalex": "W7220714518",
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.10.03.756465v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-03",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/f9c5c5pj4j87y.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:26:11Z",
      "native_id": "10.64898/2026.10.03.756465",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.10.03.756465v1"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "biorxiv:cell biology",
      "subfield": "cell-biology"
    }
  ],
  "title": "TriPort tangential flow filtration: a lab-assembled system for EV isolation across diverse cell-culture sources",
  "title_text": "TriPort tangential flow filtration: a lab-assembled system for EV isolation across diverse cell-culture sources",
  "type": "preprint",
  "updated_at": "2026-10-30T12:26:11Z",
  "url": "https://research.draconitic.com/paper/f9c5c5pj4j87y",
  "venue": "bioRxiv",
  "versions": []
}