{
  "abstract": "De novo protein binders can now be generated far faster than they can be tested. Current validation routes - display selection, cell-free two-hybrid assays, or surface-based biophysics - require cell culture, colony picking, protein purification, target labeling or sensor immobilisation, take 2 to 8 weeks, and require additional methods for affinity and kinetics characterisation. Here we report an end-to-end, cell- and purification-free pipeline that goes from gene to binding affinity and kinetics in three days without genetically modified organisms, cell culture, purification, labelling or immobilisation. An automated, metric-driven loop of RFdiffusion, ProteinMPNN and AlphaFold2 initial-guess generates and filters designs, which are ranked by Boltz-2 ipSAE, assembled by Golden Gate cloning from linear DNA, expressed in a cell-free protein synthesis lysate as fluorescent protein fusions, and screened directly in the unpurified product by flow-induced dispersion analysis (FIDA). Against human growth hormone, 11 of 30 designs (37%) bound, and the four strongest had equilibrium dissociation constants of 0.93-9.98 nM with on- and off-rates resolved from the same cell-free material. Each sample run measures expression level, hydrodynamic radius, oligomeric and aggregation state alongside the binding readout, producing the standardised, per-design datasets that model training requires.",
  "abstract_shown": true,
  "affiliations": [
    "Technical University of Denmark (DTU)"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 12,
  "authors": [
    {
      "name": "D. Ruiz Espi"
    },
    {
      "name": "G. Avelar Molina"
    },
    {
      "name": "O. Sonne Buch"
    },
    {
      "name": "J. Heyd"
    },
    {
      "name": "A. Unger"
    },
    {
      "name": "L. Winther Gormsen"
    },
    {
      "name": "K. Boldsen Lund"
    },
    {
      "name": "K. Sahni Ray"
    },
    {
      "name": "C. Williams"
    },
    {
      "name": "A. Hald"
    },
    {
      "name": "T. P. Jenkins"
    },
    {
      "affs": [
        0
      ],
      "name": "E. Rivera-de-Torre"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/4w56tahteredv.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.10.19.759856",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.10.19.759856v1.full",
  "id": "4w56tahteredv",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.10.19.759856v1",
  "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",
      "molecular biology"
    ],
    [
      "openalex",
      "1312|Protein Structure and Dynamics"
    ]
  ],
  "openalex": "W7223017854",
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.10.19.759856v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-19",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/4w56tahteredv.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:28:46Z",
      "native_id": "10.64898/2026.10.19.759856",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.10.19.759856v1"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "biorxiv:molecular biology",
      "subfield": "molecular-biology"
    }
  ],
  "title": "A rapid, automation-friendly, cell-free and purification-free pipeline for binding and kinetic characterisation of de novo designed binders with high quality standardised per-design data",
  "title_text": "A rapid, automation-friendly, cell-free and purification-free pipeline for binding and kinetic characterisation of de novo designed binders with high quality standardised per-design data",
  "type": "preprint",
  "updated_at": "2026-10-30T12:28:46Z",
  "url": "https://research.draconitic.com/paper/4w56tahteredv",
  "venue": "bioRxiv",
  "versions": []
}