{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,4]],"date-time":"2025-11-04T10:49:20Z","timestamp":1762253360936,"version":"build-2065373602"},"reference-count":82,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2020,4,16]],"date-time":"2020-04-16T00:00:00Z","timestamp":1586995200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["No.401355\/2014-4, No. 472460\/2013-7"],"award-info":[{"award-number":["No.401355\/2014-4, No. 472460\/2013-7"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["001"],"award-info":[{"award-number":["001"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["LabEx MAbImprove ANR- 10-LABX-53\u201301)."],"award-info":[{"award-number":["LabEx MAbImprove ANR- 10-LABX-53\u201301)."]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Envenoming due to Loxosceles spider bites still remains a neglected disease of particular medical concern in the Americas. To date, there is no consensus for the treatment of envenomed patients, yet horse polyclonal antivenoms are usually infused to patients with identified severe medical conditions. It is widely known that venom proteins in the 30\u201335 kDa range with sphingomyelinase D (SMasesD) activity, reproduce most of the toxic effects observed in loxoscelism. Hence, we believe that monoclonal antibody fragments targeting such toxins might pose an alternative safe and effective treatment. In the present study, starting from the monoclonal antibody LimAb7, previously shown to target SMasesD from the venom of L. intermedia and neutralize its dermonecrotic activity, we designed humanized antibody V-domains, then produced and purified as recombinant single-chain antibody fragments (scFvs). These molecules were characterized in terms of humanness, structural stability, antigen-binding activity, and venom-neutralizing potential. Throughout this process, we identified some blocking points that can impact the Abs antigen-binding activity and neutralizing capacity. In silico analysis of the antigen\/antibody amino acid interactions also contributed to a better understanding of the antibody\u2019s neutralization mechanism and led to reformatting the humanized antibody fragment which, ultimately, recovered the functional characteristics for efficient in vitro venom neutralization.<\/jats:p>","DOI":"10.3390\/toxins12040256","type":"journal-article","created":{"date-parts":[[2020,4,16]],"date-time":"2020-04-16T05:15:41Z","timestamp":1587014141000},"page":"256","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Loxoscelism: Advances and Challenges in the Design of Antibody Fragments with Therapeutic Potential"],"prefix":"10.3390","volume":"12","author":[{"given":"Sabrina","family":"Karim-Silva","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Imunoqu\u00edmica, Departamento de Patologia B\u00e1sica, Universidade Federal do Paran\u00e1,  Curitiba CEP 81531-980 PR,  Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessandra","family":"Becker-Finco","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Imunoqu\u00edmica, Departamento de Patologia B\u00e1sica, Universidade Federal do Paran\u00e1,  Curitiba CEP 81531-980 PR,  Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8367-8524","authenticated-orcid":false,"given":"Isabella Gizzi","family":"Jiacomini","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Imunoqu\u00edmica, Departamento de Patologia B\u00e1sica, Universidade Federal do Paran\u00e1,  Curitiba CEP 81531-980 PR,  Brazil"},{"name":"INRA, UMR 1282, Facult\u00e9 de Pharmacie, Universit\u00e9 de Tours, 37200 Tours, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fanny","family":"Boursin","sequence":"additional","affiliation":[{"name":"INRA, UMR 1282, Facult\u00e9 de Pharmacie, Universit\u00e9 de Tours, 37200 Tours, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arnaud","family":"Leroy","sequence":"additional","affiliation":[{"name":"IPSIT (Chiral-ist), School of Pharmacy, University Paris-Saclay, 92296 Ch\u00e2tenay-Malabry, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Magali","family":"Noiray","sequence":"additional","affiliation":[{"name":"IPSIT (Chiral-ist), School of Pharmacy, University Paris-Saclay, 92296 Ch\u00e2tenay-Malabry, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juliana","family":"de Moura","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Imunoqu\u00edmica, Departamento de Patologia B\u00e1sica, Universidade Federal do Paran\u00e1,  Curitiba CEP 81531-980 PR,  Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3211-0186","authenticated-orcid":false,"given":"Nicolas","family":"Aubrey","sequence":"additional","affiliation":[{"name":"INRA, UMR 1282, Facult\u00e9 de Pharmacie, Universit\u00e9 de Tours, 37200 Tours, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8148-5297","authenticated-orcid":false,"given":"Philippe","family":"Billiald","sequence":"additional","affiliation":[{"name":"IPSIT (Chiral-ist), School of Pharmacy, University Paris-Saclay, 92296 Ch\u00e2tenay-Malabry, France"},{"name":"MCAM, UMR MNHN-CNRS 7245, Mus\u00e9um national d\u2019Histoire naturelle, 75231 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Larissa M.","family":"Alvarenga","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Imunoqu\u00edmica, Departamento de Patologia B\u00e1sica, Universidade Federal do Paran\u00e1,  Curitiba CEP 81531-980 PR,  Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,4,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1186\/s40409-017-0097-8","article-title":"Highlights in the knowledge of brown spider toxins","volume":"23","author":"Wille","year":"2017","journal-title":"J. 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