{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,11]],"date-time":"2026-11-11T14:11:40Z","timestamp":1794406300366,"version":"4.9.0"},"reference-count":72,"publisher":"Springer Science and Business Media LLC","issue":"8053","license":[{"start":{"date-parts":[[2025,1,15]],"date-time":"2025-01-15T00:00:00Z","timestamp":1736899200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,1,15]],"date-time":"2025-01-15T00:00:00Z","timestamp":1736899200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Nature"],"published-print":{"date-parts":[[2025,3,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Snakebite envenoming remains a devastating and neglected tropical disease, claiming over 100,000 lives annually and causing severe complications and long-lasting disabilities for many more\n                    <jats:sup>1,2<\/jats:sup>\n                    . Three-finger toxins (3FTx) are highly toxic components of elapid snake venoms that can cause diverse pathologies, including severe tissue damage\n                    <jats:sup>3<\/jats:sup>\n                    and inhibition of nicotinic acetylcholine receptors, resulting in life-threatening neurotoxicity\n                    <jats:sup>4<\/jats:sup>\n                    . At present, the only available treatments for snakebites consist of polyclonal antibodies derived from the plasma of immunized animals, which have high cost and limited efficacy against 3FTxs\n                    <jats:sup>5\u20137<\/jats:sup>\n                    . Here we used deep learning methods to de novo design proteins to bind short-chain and long-chain \u03b1-neurotoxins and cytotoxins from the 3FTx family. With limited experimental screening, we obtained protein designs with remarkable thermal stability, high binding affinity and near-atomic-level agreement with the computational models. The designed proteins effectively neutralized all three 3FTx subfamilies in vitro and protected mice from a lethal neurotoxin challenge. Such potent, stable and readily manufacturable toxin-neutralizing proteins could provide the basis for safer, cost-effective and widely accessible next-generation antivenom therapeutics. Beyond snakebite, our results highlight how computational design could help democratize therapeutic discovery, particularly in resource-limited settings, by substantially reducing costs and resource requirements for the development of therapies for neglected tropical diseases.\n                  <\/jats:p>","DOI":"10.1038\/s41586-024-08393-x","type":"journal-article","created":{"date-parts":[[2025,1,15]],"date-time":"2025-01-15T11:02:13Z","timestamp":1736938933000},"page":"225-231","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":183,"title":["De novo designed proteins neutralize lethal snake venom toxins"],"prefix":"10.1038","volume":"639","author":[{"given":"Susana","family":"V\u00e1zquez Torres","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0042-6592","authenticated-orcid":false,"given":"Melisa","family":"Benard Valle","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4515-2545","authenticated-orcid":false,"given":"Stephen P.","family":"Mackessy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9273-9296","authenticated-orcid":false,"given":"Stefanie K.","family":"Menzies","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8035-4719","authenticated-orcid":false,"given":"Nicholas R.","family":"Casewell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shirin","family":"Ahmadi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-5099-3971","authenticated-orcid":false,"given":"Nick J.","family":"Burlet","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edin","family":"Muratspahi\u0107","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isaac","family":"Sappington","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Max D.","family":"Overath","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0272-6150","authenticated-orcid":false,"given":"Esperanza","family":"Rivera-de-Torre","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jann","family":"Ledergerber","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6918-5574","authenticated-orcid":false,"given":"Andreas H.","family":"Laustsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim","family":"Boddum","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9473-2912","authenticated-orcid":false,"given":"Asim K.","family":"Bera","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5487-0499","authenticated-orcid":false,"given":"Alex","family":"Kang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evans","family":"Brackenbrough","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0288-4706","authenticated-orcid":false,"given":"Iara A.","family":"Cardoso","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1676-5457","authenticated-orcid":false,"given":"Edouard P.","family":"Crittenden","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebecca J.","family":"Edge","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justin","family":"Decarreau","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert J.","family":"Ragotte","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5012-1199","authenticated-orcid":false,"given":"Arvind S.","family":"Pillai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamad","family":"Abedi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4787-5168","authenticated-orcid":false,"given":"Hannah L.","family":"Han","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0313-6248","authenticated-orcid":false,"given":"Stacey R.","family":"Gerben","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1560-6673","authenticated-orcid":false,"given":"Analisa","family":"Murray","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-0757-8432","authenticated-orcid":false,"given":"Rebecca","family":"Skotheim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lynda","family":"Stuart","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4264-5125","authenticated-orcid":false,"given":"Lance","family":"Stewart","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas J. A.","family":"Fryer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2979-5663","authenticated-orcid":false,"given":"Timothy P.","family":"Jenkins","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7896-6217","authenticated-orcid":false,"given":"David","family":"Baker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,1,15]]},"reference":[{"key":"8393_CR1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-33627-9","volume":"13","author":"GBD 2019 Snakebite Envenomation Collaborators.","year":"2022","unstructured":"GBD 2019 Snakebite Envenomation Collaborators. Global mortality of snakebite envenoming between 1990 and 2019. Nat. Commun. 13, 6160 (2022).","journal-title":"Nat. Commun."},{"key":"8393_CR2","doi-asserted-by":"publisher","first-page":"17063","DOI":"10.1038\/nrdp.2017.63","volume":"3","author":"JM Guti\u00e9rrez","year":"2017","unstructured":"Guti\u00e9rrez, J. M. et al. Snakebite envenoming. Nat. Rev. Dis. Primer 3, 17063 (2017).","journal-title":"Nat. Rev. Dis. Primer"},{"key":"8393_CR3","doi-asserted-by":"publisher","first-page":"61","DOI":"10.11604\/pamj.2023.44.61.37759","volume":"44","author":"L Royte","year":"2023","unstructured":"Royte, L. & Sawarkar, A. Snake bite\u2014cytotoxic effects of snake venom: a rare clinical image. Pan Afr. Med. J. 44, 61 (2023).","journal-title":"Pan Afr. Med. J."},{"key":"8393_CR4","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1016\/j.toxicon.2013.01.019","volume":"66","author":"CM Barber","year":"2013","unstructured":"Barber, C. M., Isbister, G. K. & Hodgson, W. C. Alpha neurotoxins. Toxicon 66, 47\u201358 (2013).","journal-title":"Toxicon"},{"key":"8393_CR5","doi-asserted-by":"publisher","first-page":"103463","DOI":"10.1016\/j.jprot.2019.103463","volume":"207","author":"A Deka","year":"2019","unstructured":"Deka, A., Gogoi, A., Das, D., Purkayastha, J. & Doley, R. Proteomics of Naja kaouthia venom from North East India and assessment of Indian polyvalent antivenom by third generation antivenomics. J. Proteomics 207, 103463 (2019).","journal-title":"J. Proteomics"},{"key":"8393_CR6","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1016\/j.toxicon.2015.03.001","volume":"99","author":"AH Laustsen","year":"2015","unstructured":"Laustsen, A. H. et al. Snake venomics of monocled cobra (Naja kaouthia) and investigation of human IgG response against venom toxins. Toxicon 99, 23\u201335 (2015).","journal-title":"Toxicon"},{"key":"8393_CR7","doi-asserted-by":"publisher","first-page":"105","DOI":"10.1016\/j.jprot.2015.02.012","volume":"120","author":"KY Tan","year":"2015","unstructured":"Tan, K. Y., Tan, C. H., Fung, S. Y. & Tan, N. H. Venomics, lethality and neutralization of Naja kaouthia (monocled cobra) venoms from three different geographical regions of Southeast Asia. J. Proteomics 120, 105\u2013125 (2015).","journal-title":"J. Proteomics"},{"key":"8393_CR8","doi-asserted-by":"publisher","first-page":"38","DOI":"10.1186\/s40409-017-0127-6","volume":"23","author":"J-P Chippaux","year":"2017","unstructured":"Chippaux, J.-P. Snakebite envenomation turns again into a neglected tropical disease! J. Venomous Anim. Toxins Incl. Trop. Dis. 23, 38 (2017).","journal-title":"J. Venomous Anim. Toxins Incl. Trop. Dis."},{"key":"8393_CR9","doi-asserted-by":"publisher","first-page":"100079","DOI":"10.1016\/j.toxcx.2021.100079","volume":"11","author":"M Hamza","year":"2021","unstructured":"Hamza, M. et al. Clinical management of snakebite envenoming: future perspectives. Toxicon X 11, 100079 (2021).","journal-title":"Toxicon X"},{"key":"8393_CR10","doi-asserted-by":"publisher","first-page":"e2326","DOI":"10.1371\/journal.pntd.0002326","volume":"7","author":"SF Stone","year":"2013","unstructured":"Stone, S. F. et al. Immune response to snake envenoming and treatment with antivenom; complement activation, cytokine production and mast cell degranulation. PLoS Negl. Trop. Dis. 7, e2326 (2013).","journal-title":"PLoS Negl. Trop. Dis."},{"key":"8393_CR11","doi-asserted-by":"publisher","first-page":"1382","DOI":"10.1016\/S0140-6736(23)00002-8","volume":"401","author":"DA Warrell","year":"2023","unstructured":"Warrell, D. A. & Williams, D. J. Clinical aspects of snakebite envenoming and its treatment in low-resource settings. Lancet 401, 1382\u20131398 (2023).","journal-title":"Lancet"},{"key":"8393_CR12","doi-asserted-by":"publisher","first-page":"e20230057","DOI":"10.1590\/1678-9199-jvatitd-2023-0057","volume":"29","author":"S Thumtecho","year":"2023","unstructured":"Thumtecho, S., Burlet, N. J., Ljungars, A. & Laustsen, A. H. Towards better antivenoms: navigating the road to new types of snakebite envenoming therapies. J. Venomous Anim. Toxins Incl. Trop. Dis. 29, e20230057 (2023).","journal-title":"J. Venomous Anim. Toxins Incl. Trop. Dis."},{"key":"8393_CR13","doi-asserted-by":"publisher","first-page":"143","DOI":"10.3390\/toxins9040143","volume":"9","author":"A Silva","year":"2017","unstructured":"Silva, A., Hodgson, W. & Isbister, G. Antivenom for neuromuscular paralysis resulting from snake envenoming. Toxins 9, 143 (2017).","journal-title":"Toxins"},{"key":"8393_CR14","doi-asserted-by":"publisher","first-page":"eadk1867","DOI":"10.1126\/scitranslmed.adk1867","volume":"16","author":"IS Khalek","year":"2024","unstructured":"Khalek, I. S. et al. Synthetic development of a broadly neutralizing antibody against snake venom long-chain \u03b1-neurotoxins. Sci. Transl. Med. 16, eadk1867 (2024).","journal-title":"Sci. Transl. Med."},{"key":"8393_CR15","doi-asserted-by":"publisher","first-page":"15561","DOI":"10.1021\/acsomega.2c00280","volume":"7","author":"AH Laustsen","year":"2022","unstructured":"Laustsen, A. H. et al. In vivo neutralization of myotoxin II, a phospholipase A2 homologue from Bothrops asper venom, using peptides discovered via phage display technology. ACS Omega 7, 15561\u201315569 (2022).","journal-title":"ACS Omega"},{"key":"8393_CR16","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-42624-5","volume":"15","author":"CV S\u00f8rensen","year":"2024","unstructured":"S\u00f8rensen, C. V. et al. Antibody-dependent enhancement of toxicity of myotoxin II from Bothrops asper. Nat. Commun. 15, 173 (2024).","journal-title":"Nat. Commun."},{"key":"8393_CR17","doi-asserted-by":"publisher","first-page":"655","DOI":"10.3389\/fimmu.2020.00655","volume":"11","author":"H Bailon Calderon","year":"2020","unstructured":"Bailon Calderon, H. et al. Development of nanobodies against hemorrhagic and myotoxic components of Bothrops atrox snake venom. Front. Immunol. 11, 655 (2020).","journal-title":"Front. Immunol."},{"key":"8393_CR18","doi-asserted-by":"publisher","first-page":"1494","DOI":"10.1021\/acs.bioconjchem.2c00220","volume":"33","author":"J Wade","year":"2022","unstructured":"Wade, J. et al. Generation of multivalent nanobody-based proteins with improved neutralization of long \u03b1-neurotoxins from elapid snakes. Bioconjugate Chem. 33, 1494\u20131504 (2022).","journal-title":"Bioconjugate Chem."},{"key":"8393_CR19","doi-asserted-by":"publisher","first-page":"e4821","DOI":"10.1002\/pro.4821","volume":"32","author":"T Tulika","year":"2023","unstructured":"Tulika, T. et al. Phage display assisted discovery of a pH\u2010dependent anti\u2010\u03b1\u2010cobratoxin antibody from a natural variable domain library. Protein Sci. 32, e4821 (2023).","journal-title":"Protein Sci."},{"key":"8393_CR20","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-36393-4","volume":"14","author":"L Ledsgaard","year":"2023","unstructured":"Ledsgaard, L. et al. Discovery and optimization of a broadly-neutralizing human monoclonal antibody against long-chain \u03b1-neurotoxins from snakes. Nat. Commun. 14, 682 (2023).","journal-title":"Nat. Commun."},{"key":"8393_CR21","doi-asserted-by":"publisher","unstructured":"Glanville, J. et al. Venom protection by broadly neutralizing antibody from a snakebite subject. Preprint at bioRxiv https:\/\/doi.org\/10.1101\/2022.09.26.507364 (2022).","DOI":"10.1101\/2022.09.26.507364"},{"key":"8393_CR22","doi-asserted-by":"publisher","first-page":"469","DOI":"10.3390\/toxins14070469","volume":"14","author":"N Alomran","year":"2022","unstructured":"Alomran, N., Chinnappan, R., Alsolaiss, J., Casewell, N. R. & Zourob, M. Exploring the utility of ssDNA aptamers directed against snake venom toxins as new therapeutics for snakebite envenoming. Toxins 14, 469 (2022).","journal-title":"Toxins"},{"key":"8393_CR23","doi-asserted-by":"publisher","first-page":"113523","DOI":"10.1016\/j.bios.2021.113523","volume":"193","author":"A Anand","year":"2021","unstructured":"Anand, A. et al. Complex target SELEX-based identification of DNA aptamers against Bungarus caeruleus venom for the detection of envenomation using a paper-based device. Biosens. Bioelectron. 193, 113523 (2021).","journal-title":"Biosens. Bioelectron."},{"key":"8393_CR24","doi-asserted-by":"publisher","first-page":"66","DOI":"10.3390\/toxins8030066","volume":"8","author":"Y-J Chen","year":"2016","unstructured":"Chen, Y.-J., Tsai, C.-Y., Hu, W.-P. & Chang, L.-S. DNA aptamers against Taiwan banded krait \u03b1-bungarotoxin recognize Taiwan cobra cardiotoxins. Toxins 8, 66 (2016).","journal-title":"Toxins"},{"key":"8393_CR25","doi-asserted-by":"publisher","first-page":"44","DOI":"10.1007\/s10517-013-2274-2","volume":"156","author":"EY Savchik","year":"2013","unstructured":"Savchik, E. Y. et al. Aptamer RA36 inhibits of human, rabbit, and rat plasma coagulation activated with thrombin or snake venom coagulases. Bull. Exp. Biol. Med. 156, 44\u201348 (2013).","journal-title":"Bull. Exp. Biol. Med."},{"key":"8393_CR26","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-35985-1","volume":"8","author":"A Dhiman","year":"2018","unstructured":"Dhiman, A. et al. Rational truncation of aptamer for cross-species application to detect krait envenomation. Sci. Rep. 8, 17795 (2018).","journal-title":"Sci. Rep."},{"key":"8393_CR27","doi-asserted-by":"publisher","first-page":"53","DOI":"10.3390\/toxins11010053","volume":"11","author":"T Jenkins","year":"2019","unstructured":"Jenkins, T. et al. Toxin neutralization using alternative binding proteins. Toxins 11, 53 (2019).","journal-title":"Toxins"},{"key":"8393_CR28","unstructured":"Romanovskis, P., Rosenberry, T. L., Cusack, B. & Spatola, A. F. in Peptides: The Wave of the Future (eds Lebl, M. & Houghten, R. A.) (Springer Netherlands, 2001)."},{"key":"8393_CR29","doi-asserted-by":"publisher","first-page":"1263","DOI":"10.1016\/j.tibtech.2021.03.003","volume":"39","author":"AH Laustsen","year":"2021","unstructured":"Laustsen, A. H., Greiff, V., Karatt-Vellatt, A., Muyldermans, S. & Jenkins, T. P. Animal immunization, in vitro display technologies, and machine learning for antibody discovery. Trends Biotechnol. 39, 1263\u20131273 (2021).","journal-title":"Trends Biotechnol."},{"key":"8393_CR30","doi-asserted-by":"publisher","first-page":"107613","DOI":"10.1016\/j.toxicon.2024.107613","volume":"239","author":"A Damsbo","year":"2024","unstructured":"Damsbo, A. et al. A comparative study of the performance of E. coli and K. phaffii for expressing \u03b1-cobratoxin. Toxicon 239, 107613 (2024).","journal-title":"Toxicon"},{"key":"8393_CR31","doi-asserted-by":"publisher","first-page":"1268","DOI":"10.1158\/0008-5472.CAN-19-2295","volume":"80","author":"I Nessler","year":"2020","unstructured":"Nessler, I. et al. Increased tumor penetration of single-domain antibody\u2013drug conjugates improves in vivo efficacy in prostate cancer models. Cancer Res. 80, 1268\u20131278 (2020).","journal-title":"Cancer Res."},{"key":"8393_CR32","doi-asserted-by":"publisher","first-page":"1089","DOI":"10.1038\/s41586-023-06415-8","volume":"620","author":"JL Watson","year":"2023","unstructured":"Watson, J. L. et al. De novo design of protein structure and function with RFdiffusion. Nature 620, 1089\u20131100 (2023).","journal-title":"Nature"},{"key":"8393_CR33","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1016\/j.toxicon.2018.03.004","volume":"146","author":"AH Laustsen","year":"2018","unstructured":"Laustsen, A. H. et al. Pros and cons of different therapeutic antibody formats for recombinant antivenom development. Toxicon 146, 151\u2013175 (2018).","journal-title":"Toxicon"},{"key":"8393_CR34","doi-asserted-by":"publisher","first-page":"17","DOI":"10.4331\/wjbc.v10.i1.17","volume":"10","author":"YN Utkin","year":"2019","unstructured":"Utkin, Y. N. Last decade update for three-finger toxins: newly emerging structures and biological activities. World J. Biol. Chem. 10, 17\u201327 (2019).","journal-title":"World J. Biol. Chem."},{"key":"8393_CR35","doi-asserted-by":"publisher","first-page":"107559","DOI":"10.1016\/j.toxicon.2023.107559","volume":"238","author":"K Kalogeropoulos","year":"2024","unstructured":"Kalogeropoulos, K. et al. A comparative study of protein structure prediction tools for challenging targets: snake venom toxins. Toxicon 238, 107559 (2024).","journal-title":"Toxicon"},{"key":"8393_CR36","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-32174-7","volume":"13","author":"M Nys","year":"2022","unstructured":"Nys, M. et al. The molecular mechanism of snake short-chain \u03b1-neurotoxin binding to muscle-type nicotinic acetylcholine receptors. Nat. Commun. 13, 4543 (2022).","journal-title":"Nat. Commun."},{"key":"8393_CR37","doi-asserted-by":"publisher","unstructured":"Sappington, I. et al. Improved protein binder design using beta-pairing targeted RFdiffusion. Preprint at bioRxiv https:\/\/doi.org\/10.1101\/2024.10.11.617496 (2024).","DOI":"10.1101\/2024.10.11.617496"},{"key":"8393_CR38","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-38328-5","volume":"14","author":"NR Bennett","year":"2023","unstructured":"Bennett, N. R. et al. Improving de novo protein binder design with deep learning. Nat. Commun. 14, 2625 (2023).","journal-title":"Nat. Commun."},{"key":"8393_CR39","doi-asserted-by":"publisher","first-page":"885","DOI":"10.1007\/s00726-018-2556-0","volume":"50","author":"G De La Rosa","year":"2018","unstructured":"De La Rosa, G., Corrales-Garc\u00eda, L. L., Rodriguez-Ruiz, X., L\u00f3pez-Vera, E. & Corzo, G. Short-chain consensus alpha-neurotoxin: a synthetic 60-mer peptide with generic traits and enhanced immunogenic properties. Amino Acids 50, 885\u2013895 (2018).","journal-title":"Amino Acids"},{"key":"8393_CR40","doi-asserted-by":"publisher","first-page":"435","DOI":"10.1038\/s41586-023-06953-1","volume":"626","author":"S V\u00e1zquez Torres","year":"2024","unstructured":"V\u00e1zquez Torres, S. et al. De novo design of high-affinity binders of bioactive helical peptides. Nature 626, 435\u2013442 (2024).","journal-title":"Nature"},{"key":"8393_CR41","doi-asserted-by":"publisher","first-page":"3373","DOI":"10.3390\/molecules26113373","volume":"26","author":"A Bekbossynova","year":"2021","unstructured":"Bekbossynova, A., Zharylgap, A. & Filchakova, O. Venom-derived neurotoxins targeting nicotinic acetylcholine receptors. Molecules 26, 3373 (2021).","journal-title":"Molecules"},{"key":"8393_CR42","doi-asserted-by":"publisher","first-page":"e4296","DOI":"10.1002\/pro.4296","volume":"31","author":"S Miersch","year":"2022","unstructured":"Miersch, S. et al. Synthetic antibodies block receptor binding and current\u2010inhibiting effects of \u03b1\u2010cobratoxin from Naja kaouthia. Protein Sci. 31, e4296 (2022).","journal-title":"Protein Sci."},{"key":"8393_CR43","doi-asserted-by":"publisher","first-page":"29594","DOI":"10.1074\/jbc.M909746199","volume":"275","author":"S Antil-Delbeke","year":"2000","unstructured":"Antil-Delbeke, S. et al. Molecular determinants by which a long chain toxin from snake venom interacts with the neuronal \u03b17-nicotinic acetylcholine receptor. J. Biol. Chem. 275, 29594\u201329601 (2000).","journal-title":"J. Biol. Chem."},{"key":"8393_CR44","doi-asserted-by":"publisher","first-page":"1512","DOI":"10.1038\/sj.emboj.7600620","volume":"24","author":"Y Bourne","year":"2005","unstructured":"Bourne, Y., Talley, T. T., Hansen, S. B., Taylor, P. & Marchot, P. Crystal structure of a Cbtx\u2013AChBP complex reveals essential interactions between snake \u03b1-neurotoxins and nicotinic receptors. EMBO J. 24, 1512\u20131522 (2005).","journal-title":"EMBO J."},{"key":"8393_CR45","doi-asserted-by":"publisher","first-page":"100123","DOI":"10.1016\/j.toxcx.2022.100123","volume":"14","author":"JJ Hiu","year":"2022","unstructured":"Hiu, J. J. & Yap, M. K. K. The myth of cobra venom cytotoxin: more than just direct cytolytic actions. Toxicon X 14, 100123 (2022).","journal-title":"Toxicon X"},{"key":"8393_CR46","doi-asserted-by":"publisher","first-page":"334","DOI":"10.3390\/toxins14050334","volume":"14","author":"HP Chong","year":"2022","unstructured":"Chong, H. P., Tan, K. Y., Liu, B.-S., Sung, W.-C. & Tan, C. H. Cytotoxicity of venoms and cytotoxins from Asiatic cobras (Naja kaouthia, Naja sumatrana, Naja atra) and neutralization by antivenoms from Thailand, Vietnam, and Taiwan. Toxins 14, 334 (2022).","journal-title":"Toxins"},{"key":"8393_CR47","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-023-39222-2","volume":"13","author":"JJ Hiu","year":"2023","unstructured":"Hiu, J. J., Fung, J. K. Y., Tan, H. S. & Yap, M. K. K. Unveiling the functional epitopes of cobra venom cytotoxin by immunoinformatics and epitope-omic analyses. Sci. Rep. 13, 12271 (2023).","journal-title":"Sci. Rep."},{"key":"8393_CR48","doi-asserted-by":"publisher","first-page":"2038","DOI":"10.1046\/j.1432-1033.2003.03580.x","volume":"270","author":"PV Dubovskii","year":"2003","unstructured":"Dubovskii, P. V., Lesovoy, D. M., Dubinnyi, M. A., Utkin, Y. N. & Arseniev, A. S. Interaction of the P\u2010type cardiotoxin with phospholipid membranes. Eur. J. Biochem. 270, 2038\u20132046 (2003).","journal-title":"Eur. J. Biochem."},{"key":"8393_CR49","doi-asserted-by":"publisher","first-page":"425","DOI":"10.3390\/toxins12070425","volume":"12","author":"F Li","year":"2020","unstructured":"Li, F., Shrivastava, I. H., Hanlon, P., Dagda, R. K. & Gasanoff, E. S. Molecular mechanism by which cobra venom cardiotoxins interact with the outer mitochondrial membrane. Toxins 12, 425 (2020).","journal-title":"Toxins"},{"key":"8393_CR50","doi-asserted-by":"publisher","first-page":"570","DOI":"10.2174\/138920312803582960","volume":"13","author":"AG Konshina","year":"2012","unstructured":"Konshina, A. G., Dubovskii, P. V. & Efremov, R. G. Structure and dynamics of cardiotoxins. Curr. Protein Pept. Sci. 13, 570\u2013584 (2012).","journal-title":"Curr. Protein Pept. Sci."},{"key":"8393_CR51","doi-asserted-by":"publisher","first-page":"386","DOI":"10.1126\/science.abb9303","volume":"371","author":"TD Kazandjian","year":"2021","unstructured":"Kazandjian, T. D. et al. Convergent evolution of pain-inducing defensive venom components in spitting cobras. Science 371, 386\u2013390 (2021).","journal-title":"Science"},{"key":"8393_CR52","unstructured":"Jensen, A. Design of Consensus Toxins and Their Use for the Discovery of Broadly Neutralizing Antibodies. PhD thesis, DTU Bioengineering (2023)."},{"key":"8393_CR53","doi-asserted-by":"publisher","first-page":"1266","DOI":"10.1021\/pr101040f","volume":"10","author":"D Petras","year":"2011","unstructured":"Petras, D. et al. Snake venomics of African spitting cobras: toxin composition and assessment of congeneric cross-reactivity of the Pan-African EchiTAb-Plus-ICP antivenom by antivenomics and neutralization approaches. J. Proteome Res. 10, 1266\u20131280 (2011).","journal-title":"J. Proteome Res."},{"key":"8393_CR54","doi-asserted-by":"crossref","unstructured":"Bartlett, K. E. et al. Dermonecrosis caused by spitting cobra snakebite results from toxin potentiation and is prevented by the repurposed drug varespladib. Proc. Natl Acad. Sci. USA 121, e2315597121 (2024).","DOI":"10.1073\/pnas.2315597121"},{"key":"8393_CR55","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1016\/j.toxicon.2016.04.049","volume":"119","author":"CM Modahl","year":"2016","unstructured":"Modahl, C. M., Mukherjee, A. K. & Mackessy, S. P. An analysis of venom ontogeny and prey-specific toxicity in the monocled cobra (Naja kaouthia). Toxicon 119, 8\u201320 (2016).","journal-title":"Toxicon"},{"key":"8393_CR56","doi-asserted-by":"publisher","first-page":"703","DOI":"10.3389\/fbioe.2020.00703","volume":"8","author":"TP Jenkins","year":"2020","unstructured":"Jenkins, T. P. & Laustsen, A. H. Cost of manufacturing for recombinant snakebite antivenoms. Front. Bioeng. Biotechnol. 8, 703 (2020).","journal-title":"Front. Bioeng. Biotechnol."},{"key":"8393_CR57","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1016\/j.it.2024.03.001","volume":"45","author":"AH Laustsen","year":"2024","unstructured":"Laustsen, A. H. Recombinant snake antivenoms get closer to the clinic. Trends Immunol. 45, 225\u2013227 (2024).","journal-title":"Trends Immunol."},{"key":"8393_CR58","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1016\/j.toxicon.2015.07.334","volume":"104","author":"AH Laustsen","year":"2015","unstructured":"Laustsen, A. H., Lohse, B., Lomonte, B., Engmark, M. & Guti\u00e9rrez, J. M. Selecting key toxins for focused development of elapid snake antivenoms and inhibitors guided by a toxicity score. Toxicon 104, 43\u201345 (2015).","journal-title":"Toxicon"},{"key":"8393_CR59","doi-asserted-by":"publisher","first-page":"964","DOI":"10.1107\/S0907444998005125","volume":"54","author":"V Nastopoulos","year":"1998","unstructured":"Nastopoulos, V., Kanellopoulos, P. N. & Tsernoglou, D. Structure of dimeric and monomeric erabutoxin a refined at 1.5 \u00c5 resolution. Acta Crystallogr. D Biol. Crystallogr. 54, 964\u2013974 (1998).","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"8393_CR60","doi-asserted-by":"publisher","first-page":"3463","DOI":"10.1080\/07391102.2017.1389662","volume":"36","author":"PV Dubovskii","year":"2018","unstructured":"Dubovskii, P. V. et al. Impact of membrane partitioning on the spatial structure of an S-type cobra cytotoxin. J. Biomol. Struct. Dyn. 36, 3463\u20133478 (2018).","journal-title":"J. Biomol. Struct. Dyn."},{"key":"8393_CR61","doi-asserted-by":"publisher","first-page":"e91232","DOI":"10.1371\/journal.pone.0091232","volume":"9","author":"JA Olsen","year":"2014","unstructured":"Olsen, J. A., Balle, T., Gajhede, M., Ahring, P. K. & Kastrup, J. S. Molecular recognition of the neurotransmitter acetylcholine by an acetylcholine binding protein reveals determinants of binding to nicotinic acetylcholine receptors. PLoS ONE 9, e91232 (2014).","journal-title":"PLoS ONE"},{"key":"8393_CR62","doi-asserted-by":"publisher","first-page":"539","DOI":"10.1038\/msb.2011.75","volume":"7","author":"F Sievers","year":"2011","unstructured":"Sievers, F. et al. Fast, scalable generation of high\u2010quality protein multiple sequence alignments using Clustal Omega. Mol. Syst. Biol. 7, 539 (2011).","journal-title":"Mol. Syst. Biol."},{"key":"8393_CR63","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1038\/s41586-022-04654-9","volume":"605","author":"L Cao","year":"2022","unstructured":"Cao, L. et al. Design of protein-binding proteins from the target structure alone. Nature 605, 551\u2013560 (2022).","journal-title":"Nature"},{"key":"8393_CR64","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1107\/S0907444909047337","volume":"66","author":"W Kabsch","year":"2010","unstructured":"Kabsch, W. XDS. Acta Crystallogr. D Biol. Crystallogr. 66, 125\u2013132 (2010).","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"8393_CR65","doi-asserted-by":"publisher","first-page":"235","DOI":"10.1107\/S0907444910045749","volume":"67","author":"MD Winn","year":"2011","unstructured":"Winn, M. D. et al. Overview of the CCP4 suite and current developments. Acta Crystallogr. D Biol. Crystallogr. 67, 235\u2013242 (2011).","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"8393_CR66","doi-asserted-by":"publisher","first-page":"658","DOI":"10.1107\/S0021889807021206","volume":"40","author":"AJ McCoy","year":"2007","unstructured":"McCoy, A. J. et al. Phaser crystallographic software. J. Appl. Crystallogr. 40, 658\u2013674 (2007).","journal-title":"J. Appl. Crystallogr."},{"key":"8393_CR67","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1107\/S0907444909052925","volume":"66","author":"PD Adams","year":"2010","unstructured":"Adams, P. D. et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr. D Biol. Crystallogr. 66, 213\u2013221 (2010).","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"8393_CR68","doi-asserted-by":"publisher","first-page":"2126","DOI":"10.1107\/S0907444904019158","volume":"60","author":"P Emsley","year":"2004","unstructured":"Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr. D Biol. Crystallogr. 60, 2126\u20132132 (2004).","journal-title":"Acta Crystallogr. D Biol. Crystallogr."},{"key":"8393_CR69","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1002\/pro.3330","volume":"27","author":"CJ Williams","year":"2018","unstructured":"Williams, C. J. et al. MolProbity: more and better reference data for improved all\u2010atom structure validation. Protein Sci. 27, 293\u2013315 (2018).","journal-title":"Protein Sci."},{"key":"8393_CR70","doi-asserted-by":"publisher","first-page":"611","DOI":"10.3389\/fphar.2020.00611","volume":"11","author":"MB Pucca","year":"2020","unstructured":"Pucca, M. B. et al. Unity makes strength: exploring intraspecies and interspecies toxin synergism between phospholipases A2 and cytotoxins. Front. Pharmacol. 11, 611 (2020).","journal-title":"Front. Pharmacol."},{"key":"8393_CR71","unstructured":"LD50 Calculator. AAT Bioquest www.aatbio.com\/tools\/ld50-calculator (2015)."},{"key":"8393_CR72","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-023-49011-6","volume":"13","author":"W Laprade","year":"2023","unstructured":"Laprade, W. et al. Machine-learning guided Venom Induced Dermonecrosis Analysis tooL: VIDAL. Sci. Rep. 13, 21662 (2023).","journal-title":"Sci. Rep."}],"container-title":["Nature"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41586-024-08393-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41586-024-08393-x","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41586-024-08393-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T17:02:03Z","timestamp":1789491723000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41586-024-08393-x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,15]]},"references-count":72,"journal-issue":{"issue":"8053","published-print":{"date-parts":[[2025,3,6]]}},"alternative-id":["8393"],"URL":"https:\/\/doi.org\/10.1038\/s41586-024-08393-x","relation":{"has-preprint":[{"id-type":"doi","id":"10.21203\/rs.3.rs-4402792\/v1","asserted-by":"object"}]},"ISSN":["0028-0836","1476-4687"],"issn-type":[{"value":"0028-0836","type":"print"},{"value":"1476-4687","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,15]]},"assertion":[{"value":"8 May 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"13 November 2024","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"15 January 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"14 September 2026","order":5,"name":"change_date","label":"Change Date","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"Update","order":6,"name":"change_type","label":"Change Type","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The version of this article initially published was available under an open access CC-BY-NC-ND license and is now available under an open access CC-BY license.","order":7,"name":"change_details","label":"Change Details","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"D.B., S.V.T., T.P.J., M.B.V., S.P.M., S.K.M., N.R.C., E.M., I.S., A.K.B. and A.K. are inventors on a provisional patent application submitted by the University of Washington for the design and composition of the proteins created in this study.","order":1,"name":"Ethics","label":"Competing interests","group":{"name":"EthicsHeading","label":"Ethics"}}]}}