{
  "abstract": "Site-selective bridging of cysteines and native disulfide bonds represents a powerful strategy for peptide and protein modification. However, most bis-electrophilic reagents rely on azide-alkyne cycloadditions for downstream bioconjugation, while existing tetrazine-based cysteine linkers can generate undesired byproducts, such as peptide cyclodimers, or incorporate additional components that are not directly involved in conjugation or ligation. Here, we report divinyl- s -tetrazine ( DvTz ) as a minimal linchpin in which the electron-deficient tetrazine core activates the vinyl groups as Michael acceptors for efficient cysteine conjugation, while simultaneously serving as a bioorthogonal handle for subsequent inverse-electron-demand Diels-Alder (IEDDA) ligation. DvTz enables high-yielding, cysteine-selective stapling of unprotected peptides, tolerates all twenty canonical amino acids, and accommodates a broad range of sequences and stapling distances. A one-pot stapling-IEDDA sequence enables facile installation of functional tags without intermediate purification. The chemistry was extended to antibody Fab fragments, achieving site-specific disulfide rebridging and subsequent IEDDA ligation across three distinct Fab proteins, with exclusive formation of singly conjugated species. A Fab-drug conjugate was constructed with retained antigen-binding activity and selective potency against HER2-positive cell lines, showcasing the application of DvTz in therapeutically relevant settings. Collectively, this work establishes DvTz as a simple and versatile linchpin for peptide and protein bioconjugation.",
  "abstract_shown": true,
  "affiliations": [
    "Yusuf Hamied Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, United Kingdom, CB2 1EW",
    "Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge, United Kingdom, CB2 1PD",
    "Engineering Biology Interdisciplinary Research Centre",
    "University of Cambridge"
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 7,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Tianyi Yang",
      "orcid": "0009-0008-9603-4411"
    },
    {
      "affs": [
        1
      ],
      "name": "Yujia Cui",
      "orcid": "0009-0004-0520-7542"
    },
    {
      "affs": [
        1
      ],
      "name": "Danye Chen"
    },
    {
      "affs": [
        0
      ],
      "name": "Daria K. Wanic"
    },
    {
      "affs": [
        0
      ],
      "name": "Linwei Zeng"
    },
    {
      "affs": [
        1,
        2,
        3
      ],
      "name": "Mark R. Howarth"
    },
    {
      "affs": [
        0
      ],
      "name": "David R. Spring"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/4m61z1gwnmyhp.bib",
  "cited_by": 0,
  "doi": "10.26434/chemrxiv.15007631/v2",
  "fulltext_url": null,
  "id": "4m61z1gwnmyhp",
  "issns": [],
  "landing_url": "https://chemrxiv.org/doi/full/10.26434/chemrxiv.15007631/v2",
  "language": null,
  "licence": "CC-BY-NC-ND",
  "licence_label": "CC BY-NC-ND",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by-nc-nd/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1605|Click Chemistry and Applications"
    ]
  ],
  "openalex": "W7223911225",
  "pdf_url": "https://chemrxiv.org/doi/pdf/10.26434/chemrxiv.15007631/v2",
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-27",
  "publisher": "American Chemical Society (ACS)",
  "ris": "https://research.draconitic.com/paper/4m61z1gwnmyhp.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:26Z",
      "native_id": "10.26434/chemrxiv.15007631",
      "source": "chemrxiv",
      "url": "https://chemrxiv.org/doi/full/10.26434/chemrxiv.15007631/v2"
    }
  ],
  "subjects": [
    {
      "field": "chemistry",
      "origin": "openalex:1605|Click Chemistry and Applications",
      "subfield": "organic"
    }
  ],
  "title": "A Minimal Tetrazine Linchpin for Cysteine Bridging and Bioorthogonal Ligation",
  "title_text": "A Minimal Tetrazine Linchpin for Cysteine Bridging and Bioorthogonal Ligation",
  "type": "preprint",
  "updated_at": "2026-10-30T12:17:26Z",
  "url": "https://research.draconitic.com/paper/4m61z1gwnmyhp",
  "venue": "ChemRxiv",
  "versions": []
}