{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:02:16Z","timestamp":1760058136597,"version":"build-2065373602"},"reference-count":31,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,3,14]],"date-time":"2025-03-14T00:00:00Z","timestamp":1741910400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005073","name":"Agency for Defense Development, Republic of Korea","doi-asserted-by":"publisher","award":["912797301"],"award-info":[{"award-number":["912797301"]}],"id":[{"id":"10.13039\/501100005073","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Botulinum neurotoxins (BoNTs) are the most potent toxins on Earth and are classified as Category A biological agents. BoNTs lead to paralysis in humans and cause botulism. Antibody therapeutics can effectively treat toxin-mediated infectious diseases. In this study, we generated a pharmaceutical humanized monoclonal antibody (HZ45 mAb) to prevent or treat botulism. HZ45 binds to the heavy chain receptor (HCR) domain of the toxin, preventing the toxin from entering the cell. The mAb was produced using hybridoma technology and phage display. We evaluated HZ45 mAb for the neutralization of BoNT serotype A (BoNT\/A) in mice and rabbits. The survival results showed that pretreatment with HZ45 mAb provided 100% protection at a dose of 0.1 mg per mouse against a maximum of 100 LD50 of BoNT\/A. To assess the therapeutic efficacy of HZ45 mAb in New Zealand white rabbits (NZWs), a 5 mg dose was administered 4 or 8 h after challenge with 10 LD50. The results indicated that 5 mg of HZ45 could treat the NZWs within 8 h after exposure to 10 LD50 botulinum. Consequently, in an in vivo context, including mice and rabbits, HZ45 mAb could protect against botulinum type A intoxication.<\/jats:p>","DOI":"10.3390\/toxins17030138","type":"journal-article","created":{"date-parts":[[2025,3,14]],"date-time":"2025-03-14T07:02:16Z","timestamp":1741935736000},"page":"138","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["An Effective Prophylactic and Therapeutic Protection Against Botulinum Type A Intoxication in Mice and Rabbits Using a Humanized Monoclonal Antibody"],"prefix":"10.3390","volume":"17","author":[{"given":"Chi Ho","family":"Yu","sequence":"first","affiliation":[{"name":"Agency for Defense Development, Yuseong, P.O. Box 35, Daejeon 305-600, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Young-Jo","family":"Song","sequence":"additional","affiliation":[{"name":"Agency for Defense Development, Yuseong, P.O. Box 35, Daejeon 305-600, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-1678-599X","authenticated-orcid":false,"given":"Dong Hyun","family":"Song","sequence":"additional","affiliation":[{"name":"Agency for Defense Development, Yuseong, P.O. Box 35, Daejeon 305-600, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hae Eun","family":"Joe","sequence":"additional","affiliation":[{"name":"Agency for Defense Development, Yuseong, P.O. Box 35, Daejeon 305-600, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chang-Hwan","family":"Kim","sequence":"additional","affiliation":[{"name":"Agency for Defense Development, Yuseong, P.O. Box 35, Daejeon 305-600, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9959-7254","authenticated-orcid":false,"given":"Hyungseok","family":"Yun","sequence":"additional","affiliation":[{"name":"Agency for Defense Development, Yuseong, P.O. Box 35, Daejeon 305-600, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Na Young","family":"Kim","sequence":"additional","affiliation":[{"name":"ABION Inc., Seoul 08394, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Euni","family":"Sim","sequence":"additional","affiliation":[{"name":"ABION Inc., Seoul 08394, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seong Tae","family":"Jeong","sequence":"additional","affiliation":[{"name":"Agency for Defense Development, Yuseong, P.O. Box 35, Daejeon 305-600, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-5214-7525","authenticated-orcid":false,"given":"Gyeung Haeng","family":"Hur","sequence":"additional","affiliation":[{"name":"Agency for Defense Development, Yuseong, P.O. Box 35, Daejeon 305-600, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1111\/j.1365-2958.2004.04377.x","article-title":"BotR\/A and TetR are alternative RNA polymerase sigma factors controlling the expression of the neurotoxin and associated protein genes in Clostridium botulinum type A and Clostridium tetani","volume":"55","author":"Raffestin","year":"2005","journal-title":"Mol. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1093\/infdis\/jit450","article-title":"Molecular characterization of a novel botulinum neurotoxin type H gene","volume":"209","author":"Dover","year":"2014","journal-title":"J. Infect. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1038\/nrmicro3295","article-title":"Botulinum neurotoxins: Genetic, structural and mechanistic insights","volume":"12","author":"Rossetto","year":"2014","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1001\/jama.285.8.1059","article-title":"Botulinum toxin as a biological weapon: Medical and public health management","volume":"285","author":"Arnon","year":"2001","journal-title":"JAMA"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"818","DOI":"10.1128\/JB.01180-06","article-title":"Genetic diversity among Botulinum Neurotoxin-producing clostridial strains","volume":"189","author":"Hill","year":"2007","journal-title":"J. Bacteriol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1146\/annurev.pharmtox.44.101802.121554","article-title":"Identification of the major steps in botulinum toxin action","volume":"44","author":"Simpson","year":"2004","journal-title":"Annu. Rev. Pharmacol. Toxicol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/S0140-6736(05)72119-X","article-title":"Extreme potency of botulinum toxin","volume":"355","author":"Schechter","year":"2000","journal-title":"Lancet"},{"key":"ref_8","first-page":"275","article-title":"Bioterrorism in Canada: An economic assessment of prevention and postattack response","volume":"12","author":"Finlay","year":"2001","journal-title":"Can. J. Infect. Dis."},{"key":"ref_9","unstructured":"(2024, May 12). Cangene Corp. BAT\u00ae [Botulism Antitoxin Heptavalent (A, B, C, D, E, F, G)\u2013(Equine)] Sterile Solution for Injection, Available online: https:\/\/www.fda.gov\/media\/85514\/download."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1056\/NEJMoa051926","article-title":"Human botulism immune globulin for the treatment of infant botulism","volume":"354","author":"Arnon","year":"2006","journal-title":"N. Engl. J. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5047","DOI":"10.1128\/AAC.02830-14","article-title":"Safety and pharmacokinetics of XOMA 3AB, a novel mixture of three monoclonal antibodies against botulinum toxin A","volume":"58","author":"Nayak","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Lu, J., Jiang, Y., Guo, J., Chen, L., Liu, F., Li, Z., Liu, X., Du, P., Yu, Y., and Wang, R. (2023). A human bispecific antibody neutralizes botulinum neurotoxin serotype A. Sci. Rep., 13.","DOI":"10.1038\/s41598-023-48008-5"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Snow, D.M., Cobb, R.R., Martinez, J., Finger-Baker, I., Collins, L., Terpening, S., Syar, E.S., Niemuth, N., Kobs, D., and Barnewall, R. (2021). A Monoclonal Antibody Combination against both Serotypes A and B Botulinum Toxin Prevents Inhalational Botulism in a Guinea Pig Model. Toxins, 13.","DOI":"10.3390\/toxins13010031"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Garcia-Rodriguez, C., Yan, S., Geren, I.N., Knopp, K.A., Dong, J., Sun, Z., Lou, J., Conrad, F., Wen, W.H., and Farr-Jones, S. (2021). A Four-Monoclonal Antibody Combination Potently Neutralizes Multiple Botulinum Neurotoxin Serotypes C and D. Toxins, 13.","DOI":"10.3390\/toxins13090641"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1016\/0002-9343(80)90469-6","article-title":"Hypersensitivity reactions associated with botulinal antitoxin","volume":"69","author":"Black","year":"1980","journal-title":"Am. J. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"570","DOI":"10.1128\/IAI.69.1.570-574.2001","article-title":"High-affinity, protective antibodies to the binding domain of botulinum neurotoxin type A","volume":"69","author":"Pless","year":"2001","journal-title":"Infect. Immun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.jmb.2005.06.003","article-title":"Molecular evolution of antibody affinity for sensitive detection of botulinum neutrotoxin type A","volume":"351","author":"Razai","year":"2005","journal-title":"J. Mol. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"446","DOI":"10.4161\/mabs.27773","article-title":"Development of neutralizing scFv-Fc against botulinum neurotoxin A light chain from a macaque immune library","volume":"6","author":"Miethe","year":"2014","journal-title":"MAbs"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4305","DOI":"10.1128\/IAI.00405-09","article-title":"Antibody protection against botulinum neurotoxin intoxication in mice","volume":"77","author":"Cheng","year":"2009","journal-title":"Infect. Immun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1006\/taap.1994.1181","article-title":"Measurement of botulinum toxin activity: Evaluation of the lethality assay","volume":"128","author":"Pearce","year":"1994","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Adekar, S.P., Takahashi, T., Jones, R.M., Al-Saleem, F.H., Ancharski, D.M., Root, M.J., Kapadnis, B.P., Simpson, L.L., and Dessain, S.K. (2008). Neutralization of botulinum neurotoxin by a human monoclonal antibody specific for the catalytic light chain. PLoS ONE, 3.","DOI":"10.1371\/journal.pone.0003023"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"991","DOI":"10.1177\/1087057109343206","article-title":"Neutralizing antibodies of botulinum neurotoxin serotype A screened from a fully synthetic human antibody phage display library","volume":"14","author":"Yu","year":"2009","journal-title":"J. Biomol. Screen."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"11346","DOI":"10.1073\/pnas.172229899","article-title":"Potent neutralization of botulinum neurotoxin by recombinant oligoclonal antibody","volume":"99","author":"Nowakowski","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Fan, Y., Garcia-Rodriguez, C., Lou, J., Wen, W., Conrad, F., Zhai, W., Smith, T.J., Smith, L.A., and Marks, J.D. (2017). A three monoclonal antibody combination potently neutralizes multiple botulinum neurotoxin serotype F subtypes. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0174187"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Garcia-Rodriguez, C., Razai, A., Geren, I.N., Lou, J., Conrad, F., Wen, W.H., Farr-Jones, S., Smith, T.J., Brown, J.L., and Skerry, J.C. (2018). A Three Monoclonal Antibody Combination Potently Neutralizes Multiple Botulinum Neurotoxin Serotype E Subtypes. Toxins, 10.","DOI":"10.3390\/toxins10030105"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1080\/21645515.2017.1405201","article-title":"A mutated recombinant subunit vaccine protects mice and guinea pigs against botulinum type A intoxication","volume":"14","author":"Yu","year":"2018","journal-title":"Hum. Vaccin. Immunother."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1002\/eji.1830060713","article-title":"Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion","volume":"6","author":"Kohler","year":"1976","journal-title":"Eur. J. Immunol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1038\/nm0103-129","article-title":"Engineered antibodies","volume":"9","author":"Hudson","year":"2003","journal-title":"Nat. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.ymeth.2005.01.004","article-title":"Use of human germline genes in a CDR homology-based approach to antibody humanization","volume":"36","author":"Hwang","year":"2005","journal-title":"Methods"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1016\/0161-5890(92)90047-2","article-title":"Possible use of similar framework region amino acid sequences between human and mouse immunoglobulins for humanizing mouse antibodies","volume":"29","author":"Wu","year":"1992","journal-title":"Mol. Immunol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"207","DOI":"10.3390\/toxins3030207","article-title":"Post-intoxication inhibition of botulinum neurotoxin serotype A within neurons by small-molecule, non-peptidic inhibitors","volume":"3","author":"Ruthel","year":"2011","journal-title":"Toxins"}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/17\/3\/138\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:53:36Z","timestamp":1760028816000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/17\/3\/138"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,14]]},"references-count":31,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["toxins17030138"],"URL":"https:\/\/doi.org\/10.3390\/toxins17030138","relation":{},"ISSN":["2072-6651"],"issn-type":[{"type":"electronic","value":"2072-6651"}],"subject":[],"published":{"date-parts":[[2025,3,14]]}}}