{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T18:55:11Z","timestamp":1790794511110,"version":"4.1.0"},"reference-count":55,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"40","funder":[{"DOI":"10.13039\/100010269","name":"Wellcome","doi-asserted-by":"publisher","award":["221702\/Z\/20\/Z"],"award-info":[{"award-number":["221702\/Z\/20\/Z"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002727","name":"Ananda Mahidol Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002727","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011958","name":"Danmarks Frie Forskningsfond","doi-asserted-by":"publisher","award":["5332-00023B"],"award-info":[{"award-number":["5332-00023B"]}],"id":[{"id":"10.13039\/501100011958","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100025276","name":"Novo Nordisk USA","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100025276","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Knut and Alice Wallenberg foundation","id":[{"id":"https:\/\/ror.org\/004hzzk67","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Villum Foundation","award":["00025302"],"award-info":[{"award-number":["00025302"]}],"id":[{"id":"https:\/\/ror.org\/05nqkay65","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Independent Research Fund Denmark","award":["grant 10.46540\/4283-00192B"],"award-info":[{"award-number":["grant 10.46540\/4283-00192B"]}],"id":[{"id":"https:\/\/ror.org\/05svhj534","id-type":"ROR","asserted-by":"crossref"}]},{"name":"European Union\u2019s Horizon","award":["850974"],"award-info":[{"award-number":["850974"]}]},{"name":"Eurotech University Allicance programme"},{"name":"Eurotech University Allicance programme"}],"content-domain":{"domain":["www.science.org"],"crossmark-restriction":true},"short-container-title":["Sci. Adv."],"published-print":{"date-parts":[[2026,10,2]]},"abstract":"<jats:p>\n                    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\u2013only antibodies (V\n                    <jats:sub>H<\/jats:sub>\n                    Hs) against long-chain \u03b1-neurotoxins. Multiplexed yeast display screening generated a dataset of diverse hits with varying binding specificities. AlphaFold3-generated V\n                    <jats:sub>H<\/jats:sub>\n                    H-toxin complex predictions enabled stringent high-precision structural triage of polyspecific V\n                    <jats:sub>H<\/jats:sub>\n                    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\n                    <jats:sub>H<\/jats:sub>\n                    Hs, with experimental validation confirming that optimized V\n                    <jats:sub>H<\/jats:sub>\n                    H variants maintained broad binding specificity across toxins. This integrated approach supports structure-guided prioritization of polyspecific V\n                    <jats:sub>H<\/jats:sub>\n                    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.\n                  <\/jats:p>","DOI":"10.1126\/sciadv.aef5325","type":"journal-article","created":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T17:58:26Z","timestamp":1790791106000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":0,"title":["Multiobjective V\n                    <sub>H<\/sub>\n                    H discovery through integrated high-throughput screening and AlphaFold3-guided structural prioritization"],"prefix":"10.1126","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-6347-9809","authenticated-orcid":true,"given":"Max D.","family":"Overath","sequence":"first","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."},{"name":"Center for Translational Protein Design, Technical University of Denmark, Kongens Lyngby, Denmark."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8853-6568","authenticated-orcid":true,"given":"Suthimon","family":"Thumtecho","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."},{"name":"Division of Toxicology, Department of Medicine, Chulalongkorn University and King Chulalongkorn Memorial Hospital, Thai Red Cross Society, 10330 Pathumwan, Bangkok, Thailand."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0272-6150","authenticated-orcid":true,"given":"Esperanza","family":"Rivera-de-Torre","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."},{"name":"Center for Translational Protein Design, Technical University of Denmark, Kongens Lyngby, Denmark."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0042-6592","authenticated-orcid":true,"given":"Melisa","family":"Benard-Valle","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9266-3017","authenticated-orcid":true,"given":"Darian S.","family":"Wolff","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."},{"name":"Center for Translational Protein Design, Technical University of Denmark, Kongens Lyngby, Denmark."},{"name":"Novonesis, Kongens Lyngby, Denmark."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1904-1163","authenticated-orcid":true,"given":"Rahmat","family":"Grahadi","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."},{"name":"Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Brawijaya, Malang, Indonesia."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1339-8620","authenticated-orcid":true,"given":"Kasper H.","family":"Bj\u00f6rnsson","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."},{"name":"Center for Translational Protein Design, Technical University of Denmark, Kongens Lyngby, Denmark."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas S. H.","family":"Rygaard","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."},{"name":"Center for Translational Protein Design, Technical University of Denmark, Kongens Lyngby, Denmark."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-4639-6161","authenticated-orcid":true,"given":"Nils","family":"Hofmann","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-9774-095X","authenticated-orcid":true,"given":"Jann","family":"Ledergerber","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."},{"name":"Molecular Design Laboratory, ETH Zurich, Zurich, Switzerland."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2158-0601","authenticated-orcid":true,"given":"Anne","family":"Ljungars","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6918-5574","authenticated-orcid":true,"given":"Andreas H.","family":"Laustsen","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3927-7897","authenticated-orcid":true,"given":"Simon","family":"Olsson","sequence":"additional","affiliation":[{"name":"Department of Computer Science and Engineering, Chalmers University of Technology and University of Gothenburg, Gothenburg, Sweden."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4338-0405","authenticated-orcid":true,"given":"Thomas J.","family":"Fryer","sequence":"additional","affiliation":[{"name":"IMES, Massachusetts Institute of Technology, Cambridge, MA, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2979-5663","authenticated-orcid":true,"given":"Timothy P.","family":"Jenkins","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark."},{"name":"Center for Translational Protein Design, Technical University of Denmark, Kongens Lyngby, Denmark."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1093\/abt\/tbac021"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.compchemeng.2024.108585"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1080\/19420862.2023.2171248"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-39022-2"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2021.03.003"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.4331\/wjbc.v3.i12.187"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-33627-9"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0005361"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-025-09661-0"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-36393-4"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.adk1867"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-48539-z"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-06086-4"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2018.03.004"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2020.00703"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0012833"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jprot.2015.02.002"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2015.03.001"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1111\/jnc.13975"},{"key":"e_1_3_2_21_2","unstructured":"D. A. Warrell \u201cClinical toxicology of snakebite in Asia\u201d in Handbook of Clinical Toxicology of Animal Venoms and Poisons (CRC Press 2008)."},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4296"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0069495"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-08393-x"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2022.106991"},{"key":"e_1_3_2_26_2","first-page":"S0167-7799(26)0","article-title":"One-pot expression of an oligoclonal nanobody-based antivenom in growth-decoupled Escherichia coli","author":"Stargardt P.","year":"2026","unstructured":"P. Stargardt, A. Ljungars, D. Scheich, N. Lingg, E. S. Elmose, S. Ahmadi, F. Simon, A. Jungbauer, J. Mairhofer, A. H. Laustsen, One-pot expression of an oligoclonal nanobody-based antivenom in growth-decoupled Escherichia coli. Trends Biotechnol., S0167-7799(26)00085-5 (2026).","journal-title":"Trends Biotechnol."},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0049535"},{"key":"e_1_3_2_28_2","doi-asserted-by":"crossref","unstructured":"F. Ferrara M. F. Soluri D. Sblattero \u201cRecombinant antibody selections by combining phage and yeast display\u201d in Human Monoclonal Antibodies: Methods and Protocols M. Steinitz Ed. (Springer New York NY 2019) pp. 339\u2013352; https:\/\/doi.org\/10.1007\/978-1-4939-8958-4_16.","DOI":"10.1007\/978-1-4939-8958-4_16"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.6b02334"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2015.09.006"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkz131"},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-35668-6"},{"key":"e_1_3_2_33_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41551-021-00699-9"},{"key":"e_1_3_2_34_2","doi-asserted-by":"publisher","DOI":"10.1080\/19420862.2022.2069075"},{"key":"e_1_3_2_35_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-31457-3"},{"key":"e_1_3_2_36_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2023.1249247"},{"key":"e_1_3_2_37_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cels.2025.101391"},{"key":"e_1_3_2_38_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07487-w"},{"key":"e_1_3_2_39_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.adk8946"},{"key":"e_1_3_2_40_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2014.09.026"},{"key":"e_1_3_2_41_2","doi-asserted-by":"publisher","DOI":"10.1038\/s42004-025-01600-4"},{"key":"e_1_3_2_42_2","unstructured":"EMBL-EBI AlphaFold: Confidence scores in AlphaFold-Multimer; www.ebi.ac.uk\/training\/online\/courses\/alphafold\/inputs-and-outputs\/evaluating-alphafolds-predicted-structures-using-confidence-scores\/confidence-scores-in-alphafold-multimer\/."},{"key":"e_1_3_2_43_2","doi-asserted-by":"crossref","unstructured":"L. S. Mille-Fragoso J. N. Wang C. L. Driscoll H. Dai T. Widatalla X. Zhang B. L. Hie X. J. Gao Efficient generation of epitope-targeted de novo antibodies with Germinal. bioRxiv 677421 [Preprint] (2025); https:\/\/doi.org\/10.1101\/2025.09.19.677421.","DOI":"10.1038\/s41587-026-03187-0"},{"key":"e_1_3_2_44_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-025-09721-5"},{"key":"e_1_3_2_45_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7600620"},{"key":"e_1_3_2_46_2","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aef7163"},{"key":"e_1_3_2_47_2","first-page":"1","article-title":"Massive sampling strategy for antibody\u2013antigen targets in CAPRI Round 55 with MassiveFold","volume":"2025","author":"Raouraoua N.","year":"2025","unstructured":"N. Raouraoua, M. F. Lensink, G. Brysbaert, Massive sampling strategy for antibody\u2013antigen targets in CAPRI Round 55 with MassiveFold. Proteins 2025, 1\u20139 (2025).","journal-title":"Proteins"},{"key":"e_1_3_2_48_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btad573"},{"key":"e_1_3_2_49_2","doi-asserted-by":"crossref","unstructured":"E. Smorodina M. Ali K. K. Brumat L. Salicari S. Miklavc A. Kappassov C. Fu P. Sormanni A. de Marco V. Greiff Structural plausibility without binding specificity: Limits of AI-based antibody-antigen structure prediction confidence scores. bioRxiv 709004 [Preprint] (2026); https:\/\/doi.org\/10.64898\/2026.03.02.709004.","DOI":"10.64898\/2026.03.02.709004"},{"key":"e_1_3_2_50_2","doi-asserted-by":"crossref","unstructured":"Y. Zhang C. Gong J. Sun J. Guan M. Ren S. Xue H. Zhang W. Ma Z. Liu X. Chen W. Xiao Protenix-v2: Broadening the reach of structure prediction and biomolecular design. bioRxiv 717613 [Preprint] (2026); https:\/\/doi.org\/10.64898\/2026.04.10.717613.","DOI":"10.64898\/2026.04.10.717613"},{"key":"e_1_3_2_51_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2007.13"},{"key":"e_1_3_2_52_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-09678-w"},{"key":"e_1_3_2_53_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.3364"},{"key":"e_1_3_2_54_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-27794-3"},{"key":"e_1_3_2_55_2","doi-asserted-by":"publisher","DOI":"10.1080\/19420862.2022.2085536"},{"key":"e_1_3_2_56_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(01)00461-5"}],"container-title":["Science Advances"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/sciadv.aef5325","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T17:58:35Z","timestamp":1790791115000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/sciadv.aef5325"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,2]]},"references-count":55,"journal-issue":{"issue":"40","published-print":{"date-parts":[[2026,10,2]]}},"alternative-id":["10.1126\/sciadv.aef5325"],"URL":"https:\/\/doi.org\/10.1126\/sciadv.aef5325","relation":{},"ISSN":["2375-2548"],"issn-type":[{"value":"2375-2548","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,2]]},"assertion":[{"value":"2026-01-18","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-20","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-30","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"eaef5325"}}