{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,5]],"date-time":"2026-06-05T00:06:37Z","timestamp":1780617997803,"version":"3.54.1"},"reference-count":31,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2009,7,1]],"date-time":"2009-07-01T00:00:00Z","timestamp":1246406400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Infect Immun"],"published-print":{"date-parts":[[2009,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>The botulinum neurotoxins (BoNTs) are a large family of extremely potent, neuroparalytic, dichain proteins which act at the peripheral nervous system. The wide genetic diversity observed with this neurotoxin family poses a significant challenge for the development of an effective botulinum vaccine. The present study describes a vaccine development platform based on protein fragments representing the N-terminal two-thirds of each toxin molecule. These fragments, designated LH<jats:sub>N<\/jats:sub>, comprise the light chain and translocation domains of each neurotoxin and are devoid of any neuron-binding activity. Using codon-optimized genes, LH<jats:sub>N<\/jats:sub>fragments derived from BoNT serotypes A and B were expressed in<jats:italic>Escherichia coli<\/jats:italic>in high yield with &gt;1 g of purified, soluble fragment recoverable from 4.5 liter-scale fermentations. The protective efficacy of LH<jats:sub>N<\/jats:sub>\/A was significantly enhanced by treatment with formaldehyde, which induced intramolecular cross-linking but virtually no aggregation of the fragment. A single immunization of the modified fragment protected mice from challenge with a 10<jats:sup>3<\/jats:sup>50% lethal dose (LD<jats:sub>50<\/jats:sub>) of BoNT\/A<jats:sub>1<\/jats:sub>with an 50% effective dose (ED<jats:sub>50<\/jats:sub>) of 50 ng of the vaccine. In similar experiments, the LH<jats:sub>N<\/jats:sub>\/A vaccine was shown to protect mice against challenge with BoNT\/A subtypes A<jats:sub>1<\/jats:sub>, A<jats:sub>2<\/jats:sub>, and A<jats:sub>3<\/jats:sub>, which is the first demonstration of single-dose protection by a vaccine against the principal toxin subtypes of BoNT\/A. The LH<jats:sub>N<\/jats:sub>\/B vaccine was also highly efficacious, giving an ED<jats:sub>50<\/jats:sub>of \u223c140 ng to a challenge of 10<jats:sup>3<\/jats:sup>LD<jats:sub>50<\/jats:sub>of BoNT\/B<jats:sub>1<\/jats:sub>. In addition, LH<jats:sub>N<\/jats:sub>\/B provided single-dose protection in mice against BoNT\/B<jats:sub>4<\/jats:sub>(nonproteolytic toxin subtype).<\/jats:p>","DOI":"10.1128\/iai.01252-08","type":"journal-article","created":{"date-parts":[[2009,4,28]],"date-time":"2009-04-28T01:23:00Z","timestamp":1240881780000},"page":"2795-2801","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":34,"title":["Bivalent Recombinant Vaccine for Botulinum Neurotoxin Types A and B Based on a Polypeptide Comprising Their Effector and Translocation Domains That Is Protective against the Predominant A and B Subtypes"],"prefix":"10.1128","volume":"77","author":[{"given":"Clifford","family":"Shone","sequence":"first","affiliation":[{"name":"Health Protection Agency, Porton Down, Salisbury, Wilts SP4 0JG, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heidi","family":"Agostini","sequence":"additional","affiliation":[{"name":"Emergent BioSolutions, Inc., 300 Professional Drive, Gaithersburg, Maryland 20879"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joanna","family":"Clancy","sequence":"additional","affiliation":[{"name":"Emergent BioSolutions, Inc., 300 Professional Drive, Gaithersburg, Maryland 20879"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mili","family":"Gu","sequence":"additional","affiliation":[{"name":"Emergent BioSolutions, Inc., 300 Professional Drive, Gaithersburg, Maryland 20879"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huei-Hsiung","family":"Yang","sequence":"additional","affiliation":[{"name":"Emergent BioSolutions, Inc., 300 Professional Drive, Gaithersburg, Maryland 20879"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanfang","family":"Chu","sequence":"additional","affiliation":[{"name":"Emergent BioSolutions, Inc., 300 Professional Drive, Gaithersburg, Maryland 20879"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Virginia","family":"Johnson","sequence":"additional","affiliation":[{"name":"Emergent BioSolutions, Inc., 300 Professional Drive, Gaithersburg, Maryland 20879"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Makie","family":"Taal","sequence":"additional","affiliation":[{"name":"Health Protection Agency, Porton Down, Salisbury, Wilts SP4 0JG, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joanna","family":"McGlashan","sequence":"additional","affiliation":[{"name":"Health Protection Agency, Porton Down, Salisbury, Wilts SP4 0JG, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Brehm","sequence":"additional","affiliation":[{"name":"Health Protection Agency, Porton Down, Salisbury, Wilts SP4 0JG, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaomi","family":"Tong","sequence":"additional","affiliation":[{"name":"Emergent BioSolutions, Inc., 300 Professional Drive, Gaithersburg, Maryland 20879"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2006.07.040"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1001\/jama.285.8.1059"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0300-9084(00)01173-1"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.10.4817-4822.1998"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1046-5928(02)00002-5"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/307457a0"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1123654"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.8.4629"},{"key":"e_1_3_2_10_2","doi-asserted-by":"crossref","first-page":"697","DOI":"10.4049\/jimmunol.90.5.697","volume":"90","year":"1963","unstructured":"Fiock, M., M. 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