{
  "abstract": "Human epidermal growth factor receptor 2 (HER2) is an oncogenic receptor tyrosine kinase in breast cancer and other malignancies. A subset of HER2-positive tumours expresses 611-CTF-p95HER2, a tumour-specific, hyperactive truncated isoform associated with metastasis and treatment resistance that lacks most of the extracellular domain targeted by conventional HER2-directed antibodies. We previously developed NAZ-mAb (formerly known as Oslo-2), a monoclonal antibody against 611-CTF-p95HER2. Here, we describe a computational antibody-engineering workflow for designing variants of NAZ-mAb. Starting from the sequence alone, we modelled the NAZ-mAb-611-CTF-p95HER2 complex, generated a combinatorial mutational landscape using FoldX 5.0, and prioritized candidate variants using predicted interaction energy and developability criteria. Two variants representing distinct design strategies were selected for validation: an aromatic double mutant, NAZ-mAb v1 (L:S31W/L:H107W), and a conservative single mutant, NAZ-mAb v2 (L:S31M). Both variants were successfully expressed as recombinant IgGs; NAZ-mAb v2 achieved a five-fold higher recombinant expression yield than parental NAZ-mAb, while both variants retained antigen binding with a higher apparent signal than the parental antibody in indirect ELISA. However, Biacore two-state kinetic analysis revealed weaker affinities than the parental antibody (KD NAZ-mAb v1: 326.1 nM, NAZ-mAb v2: 94.5 nM vs parental NAZ-mAb: 53.3 nM). These findings show that the computational workflow can generate antigen-engaging NAZ-mAb variants, while also highlighting the limitations of fixed-backbone interaction-energy ranking as a predictor of binding affinity and yield. This study provides a practical framework and a cautionary benchmark for computationally driven, developability-aware antibody engineering in the absence of experimental structural data.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Immunology, University of Oslo and Oslo University Hospital, Oslo, Norway.",
    "Department of Cancer Immunology, Institute for Cancer Research, Oslo University Hospital, Oslo, Norway.",
    "Department of Clinical Cancer Research, Oslo University Hospital, Oslo, Norway.",
    "Institute of Clinical Medicine, University of Oslo, Norway.",
    "Immunoquest Therapeutics AS, Oslo, Norway."
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 4,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Puneet Rawat",
      "orcid": "0000-0002-3822-8081"
    },
    {
      "affs": [
        1,
        2,
        3
      ],
      "name": "Jon Amund Kyte",
      "orcid": "0000-0002-2854-3694"
    },
    {
      "affs": [
        0
      ],
      "name": "Victor Greiff",
      "orcid": "0000-0003-2622-5032"
    },
    {
      "affs": [
        1,
        4
      ],
      "name": "Esmaeil Dorraji"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/4wck5kn0f7pjs.bib",
  "cited_by": 0,
  "doi": "10.1016/j.crstbi.2026.100199",
  "fulltext_url": "https://europepmc.org/articles/PMC13683858",
  "id": "4wck5kn0f7pjs",
  "issns": [
    "2665-928X"
  ],
  "landing_url": "https://europepmc.org/article/MED/42909185",
  "language": "eng",
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "2741|Monoclonal and Polyclonal Antibodies Research"
    ]
  ],
  "openalex": "W7221680369",
  "pdf_url": "https://europepmc.org/articles/PMC13683858?pdf=render",
  "pmcid": "PMC13683858",
  "pmid": "42909185",
  "published": "2026-10-13",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/4wck5kn0f7pjs.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:20:10Z",
      "native_id": "MED:42909185",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42909185"
    }
  ],
  "subjects": [
    {
      "field": "medicine",
      "origin": "openalex:2741|Monoclonal and Polyclonal Antibodies Research",
      "subfield": "imaging"
    }
  ],
  "title": "Structure-guided computational design and mechanistic understanding of the p95HER2-targeting NAZ-mAb antibody and its variants.",
  "title_text": "Structure-guided computational design and mechanistic understanding of the p95HER2-targeting NAZ-mAb antibody and its variants.",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:20:10Z",
  "url": "https://research.draconitic.com/paper/4wck5kn0f7pjs",
  "venue": "Current research in structural biology",
  "versions": []
}