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However, designing and developing broadly\u2010neutralizing antibodies can be cumbersome using traditional low\u2010throughput iterative protein engineering methods. Here, we present a new high\u2010throughput approach for the standardized discovery of broadly\u2010neutralizing monoclonal antibodies relying on phage display technology and consensus antigens representing average sequences of related proteins. We showcase the utility of this approach by applying it to toxic sphingomyelinases from the venoms of species from very distant orders of the animal kingdom, the recluse spider and Gadim scorpion. First, we designed a consensus sphingomyelinase and performed three rounds of phage display selection, followed by DELFIA\u2010based screening and ranking, and benchmarked this to a similar campaign involving cross\u2010panning against recombinant versions of the native toxins. Second, we identified two scFvs that not only bind the consensus toxins, but which can also neutralize sphingomyelinase activity of native whole venom in vitro. Finally, we conclude that the phage display campaign involving the use of the consensus toxin was more successful in yielding cross\u2010neutralizing scFvs than the phage display campaign involving cross\u2010panning.<\/jats:p>","DOI":"10.1002\/pro.4901","type":"journal-article","created":{"date-parts":[[2024,2,15]],"date-time":"2024-02-15T08:35:29Z","timestamp":1707986129000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Discovery of broadly\u2010neutralizing antibodies against brown recluse spider and Gadim scorpion sphingomyelinases using consensus toxins as antigens"],"prefix":"10.1002","volume":"33","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0272-6150","authenticated-orcid":false,"given":"Esperanza","family":"Rivera\u2010de\u2010Torre","sequence":"first","affiliation":[{"name":"Department of Biotechnology and Biomedicine Technical University of Denmark  Kongens Lyngby 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