{
  "abstract": "Finding therapeutic antibodies that bind multiple related targets with high affinity and favorable biophysical properties remains challenging and resource-intensive. For snakebite antivenoms, this challenge is critical as treatments must neutralize toxins across multiple snake species. We developed a pipeline combining high-throughput yeast screening, deep sequencing, and AlphaFold3 structure prediction to identify polyspecific variable domains of heavy chain–only antibodies (V H Hs) against long-chain α-neurotoxins. Multiplexed yeast display screening generated a dataset of diverse hits with varying binding specificities. AlphaFold3-generated V H H-toxin complex predictions enabled stringent high-precision structural triage of polyspecific V H Hs that bind conserved epitopes across multiple toxins, with a representative subset experimentally confirmed to block toxin binding to the acetylcholine receptor. These structural insights provided a starting point for computational optimization of affinity and soluble expression of the V H Hs, with experimental validation confirming that optimized V H H variants maintained broad binding specificity across toxins. This integrated approach supports structure-guided prioritization of polyspecific V H H hits, offering a high-precision filter that could reduce future reliance on extensive experimental specificity screening and providing a framework applicable to other therapeutic contexts, in which substantial antigen variation occurs and broad neutralization is essential.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark.",
    "Center for Translational Protein Design, Technical University of Denmark, Kongens Lyngby, Denmark.",
    "Division of Toxicology, Department of Medicine, Chulalongkorn University and King Chulalongkorn Memorial Hospital, Thai Red Cross Society, 10330 Pathumwan, Bangkok, Thailand.",
    "Novonesis, Kongens Lyngby, Denmark.",
    "Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Brawijaya, Malang, Indonesia.",
    "Molecular Design Laboratory, ETH Zurich, Zurich, Switzerland.",
    "Department of Computer Science and Engineering, Chalmers University of Technology and University of Gothenburg, Gothenburg, Sweden.",
    "IMES, Massachusetts Institute of Technology, Cambridge, MA, USA."
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 15,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Max D. Overath",
      "orcid": "0009-0009-6347-9809"
    },
    {
      "affs": [
        0,
        2
      ],
      "name": "Suthimon Thumtecho",
      "orcid": "0000-0002-8853-6568"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Esperanza Rivera-de-Torre",
      "orcid": "0000-0002-0272-6150"
    },
    {
      "affs": [
        0
      ],
      "name": "Melisa Benard-Valle",
      "orcid": "0000-0002-0042-6592"
    },
    {
      "affs": [
        0,
        1,
        3
      ],
      "name": "Darian S. Wolff",
      "orcid": "0000-0002-9266-3017"
    },
    {
      "affs": [
        0,
        4
      ],
      "name": "Rahmat Grahadi",
      "orcid": "0000-0003-1904-1163"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Kasper H. Björnsson",
      "orcid": "0000-0003-1339-8620"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Andreas S. H. Rygaard"
    },
    {
      "affs": [
        0
      ],
      "name": "Nils Hofmann",
      "orcid": "0009-0006-4639-6161"
    },
    {
      "affs": [
        0,
        5
      ],
      "name": "Jann Ledergerber",
      "orcid": "0009-0004-9774-095X"
    },
    {
      "affs": [
        0
      ],
      "name": "Anne Ljungars",
      "orcid": "0000-0002-2158-0601"
    },
    {
      "affs": [
        0
      ],
      "name": "Andreas H. Laustsen",
      "orcid": "0000-0001-6918-5574"
    },
    {
      "affs": [
        6
      ],
      "name": "Simon Olsson",
      "orcid": "0000-0002-3927-7897"
    },
    {
      "affs": [
        7
      ],
      "name": "Thomas J. Fryer",
      "orcid": "0000-0003-4338-0405"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Timothy P. Jenkins",
      "orcid": "0000-0003-2979-5663"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/e14nkse1ky1mc.bib",
  "cited_by": 0,
  "doi": "10.1126/sciadv.aef5325",
  "fulltext_url": null,
  "id": "e14nkse1ky1mc",
  "issns": [
    "2375-2548"
  ],
  "landing_url": "https://www.science.org/doi/10.1126/sciadv.aef5325",
  "language": null,
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "2741|Monoclonal and Polyclonal Antibodies Research"
    ]
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  "openalex": "W7214886635",
  "pdf_url": null,
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-02",
  "publisher": "American Association for the Advancement of Science (AAAS)",
  "ris": "https://research.draconitic.com/paper/e14nkse1ky1mc.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:41Z",
      "native_id": "10.1126/sciadv.aef5325",
      "source": "crossref",
      "url": "https://www.science.org/doi/10.1126/sciadv.aef5325"
    }
  ],
  "subjects": [
    {
      "field": "medicine",
      "origin": "openalex:2741|Monoclonal and Polyclonal Antibodies Research",
      "subfield": "imaging"
    }
  ],
  "title": "Multiobjective V H H discovery through integrated high-throughput screening and AlphaFold3-guided structural prioritization",
  "title_text": "Multiobjective V H H discovery through integrated high-throughput screening and AlphaFold3-guided structural prioritization",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:17:41Z",
  "url": "https://research.draconitic.com/paper/e14nkse1ky1mc",
  "venue": "Science Advances",
  "versions": []
}