{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T03:43:54Z","timestamp":1773287034477,"version":"3.50.1"},"reference-count":41,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2001,7,1]],"date-time":"2001-07-01T00:00:00Z","timestamp":993945600000},"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":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2001,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p><jats:italic>Clostridium botulinum<\/jats:italic>neurotoxin type A (BTx-A) is known to inhibit the release of acetylcholine at the neuromuscular junctions and synapses and to cause neuroparalysis and death. In this study, we have identified two monoclonal antibodies, BT57-1 and BT150-3, which protect ICR mice against lethal doses of BTx-A challenge. The neutralizing activities for BT57-1 and BT150-3 were 10<jats:sup>3<\/jats:sup>and 10<jats:sup>4<\/jats:sup>times the 50% lethal dose, respectively. Using immunoblotting analysis, BT57-1 was recognized as a light chain and BT150-3 was recognized as a heavy chain of BTx-A. Also, applying the phage display method, we investigated the antibodies' neutralizing B-cell epitopes. These immunopositive phage clones displayed consensus motifs, Asp-Pro-Leu for BT57-1 and Cys-X-Asp-Cys for BT150. The synthetic peptide P4M (KGTFDPLQEPRT) corresponded to the phage-displayed peptide selected by BT57-1 and was able to bind the antibodies specifically. This peptide was also shown by competitive inhibition assay to be able to inhibit phage clone binding to BT57-1. Aspartic acid (D<jats:sup>5<\/jats:sup>) in P4M was crucial to the binding of P4M to BT57-1, since its binding activity dramatically decreased when it was changed to lysine (K<jats:sup>5<\/jats:sup>). Finally, immunizing mice with the selected phage clones elicited a specific humoral response against BTx-A. These results suggest that phage-displayed random-peptide libraries are useful in identifying the neutralizing epitopes of monoclonal antibodies. In the future, the identification of the neutralizing epitopes of BTx-A may provide important information for the identification of the BTx-A receptor and the design of a BTx-A vaccine.<\/jats:p>","DOI":"10.1128\/aem.67.7.3201-3207.2001","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T10:00:58Z","timestamp":1027764058000},"page":"3201-3207","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":28,"title":["Characterization of Neutralizing Antibodies and Identification of Neutralizing Epitope Mimics on the<i>Clostridium botulinum<\/i>Neurotoxin Type A"],"prefix":"10.1128","volume":"67","author":[{"given":"Han-Chung","family":"Wu","sequence":"first","affiliation":[{"name":"Institute of Preventive Medicine, National Defense Medical Center, San-Hsia, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chia-Tsui","family":"Yeh","sequence":"additional","affiliation":[{"name":"Institute of Preventive Medicine, National Defense Medical Center, San-Hsia, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yue-Ling","family":"Huang","sequence":"additional","affiliation":[{"name":"Institute of Preventive Medicine, National Defense Medical Center, San-Hsia, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lih-Jeng","family":"Tarn","sequence":"additional","affiliation":[{"name":"Institute of Preventive Medicine, National Defense Medical Center, San-Hsia, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chien-Cheng","family":"Lung","sequence":"additional","affiliation":[{"name":"Institute of Preventive Medicine, National Defense Medical Center, San-Hsia, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.279.5349.377"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01886751"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.278.5340.1125"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm0396-299"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(17)42071-0"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(87)80562-8"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1006\/prep.2000.1200"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.7542403"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/0167-7799(94)90137-6"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/0041-0101(84)90085-0"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291X(72)90350-6"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(93)90148-V"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1994.tb06501.x"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1001\/jama.1997.03550050061035"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/mr.46.1.86-94.1982"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(91)90354-9"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.71.5.3879-3885.1997"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1038\/256495a0"},{"key":"e_1_3_2_20_2","first-page":"883","article-title":"Identification of receptor ligands with phage display peptide libraries","volume":"40","author":"Koivunen E.","year":"1999","unstructured":"Koivunen E. 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