Structural mechanisms behind the neutralisation of long-chain α-neurotoxins by broadly neutralising VHHs discovered using a consensus antigen
Snakebite envenoming, a neglected tropical disease, affects millions globally, causing significant morbidity and mortality. Developing broadly neutralising monoclonal antibodies offers a promising approach to address the antigenic variation present in snake venoms. In this study, we designed a long-chain consensus α-neurotoxin, LCC, to serve as an antigen in a phage display-based antibody discovery campaign. Utilising a yeast expression system, we expressed LCC and identified 21 variable domains of heavy-chain-only antibodies (V<sub>H</sub>Hs) from immune libraries. These V<sub>H</sub>Hs were assessed for their binding affinity to various long-chain α-neurotoxins and their neutralising capability in vitro. The V<sub>H</sub>H with the broadest cross-reactivity and highest affinity, TPL1158_01_C09, was co-crystallised with α-cobratoxin to elucidate its binding mechanism. In vivo rodent studies demonstrated the neutralisation potential of TPL1158_01_C09. Our findings highlight that the use of a consensus toxin as an antigen coincided with the discovery of broadly neutralising V<sub>H</sub>Hs against snake venom toxins.
Authors
- Johannes R. Loeffler (ORCID: https://orcid.org/0000-0002-5724-655X)
- Alid Guadarrama-Martínez (ORCID: https://orcid.org/0009-0007-6378-0859)
- Melisa Bénard-Valle (ORCID: https://orcid.org/0000-0002-0042-6592)
- Tulika Tulika
- Monica L. Fernández‐Quintero (ORCID: https://orcid.org/0000-0002-6811-6283)
- Jens Preben Morth (ORCID: https://orcid.org/0000-0003-4077-0192)
- Andreas H. Laustsen (ORCID: https://orcid.org/0000-0001-6918-5574)
- Kim Boddum (ORCID: https://orcid.org/0000-0002-1588-9055)
- Anna Jensen (ORCID: https://orcid.org/0000-0001-8601-518X)
- Esperanza Rivera‐de‐Torre (ORCID: https://orcid.org/0000-0002-0272-6150)
- Max D. Overath (ORCID: https://orcid.org/0009-0009-6347-9809)
- Nick J. Burlet (ORCID: https://orcid.org/0009-0002-5099-3971)
- Anneline Vlamynck
- Ida Bisbo
- Alejandro Alagón (ORCID: https://orcid.org/0000-0003-1721-3726)
- César Villalobos (ORCID: https://orcid.org/0009-0007-8623-3508)
Institutions
- Scripps Research Institute (US)
- Universidad Nacional Autónoma de México (MX)
- Technical University of Denmark (DK)
Publication Details
- Journal
- Communications Chemistry
- Published
- 2025-07-19
- DOI
- https://doi.org/10.1038/s42004-025-01600-4
- Citations
- 4
- Primary Topic
- Venomous Animal Envenomation and Studies
- Type
- article
- Field-Weighted Citation Impact
- 7.27
Funders
- Villum Fonden
- European Commission
- VINNOVA
- Svenska Forskningsrådet Formas
- Consejo Nacional de Ciencia y Tecnología
- Vetenskapsrådet