{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,23]],"date-time":"2026-01-23T06:06:37Z","timestamp":1769148397956,"version":"3.49.0"},"reference-count":127,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2011,5,13]],"date-time":"2011-05-13T00:00:00Z","timestamp":1305244800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Specific treatment is not available for human botulism. Current remedial mainstay is the passive administration of polyclonal antibody to botulinum neurotoxin (BoNT) derived from heterologous species (immunized animal or mouse hybridoma) together with supportive and symptomatic management. The antibody works extracellularly, probably by blocking the binding of receptor binding (R) domain to the neuronal receptors; thus inhibiting cellular entry of the holo-BoNT. The antibody cannot neutralize the intracellular toxin. Moreover, a conventional antibody with relatively large molecular size (150 kDa) is not accessible to the enzymatic groove and, thus, cannot directly inhibit the BoNT zinc metalloprotease activity. Recently, a 15\u201320 kDa single domain antibody (VHH) that binds specifically to light chain of BoNT serotype A was produced from a humanized-camel VH\/VHH phage display library. The VHH has high sequence homology (&gt;80%) to the human VH and could block the enzymatic activity of the BoNT. Molecular docking revealed not only the interface binding between the VHH and the toxin but also an insertion of the VHH CDR3 into the toxin enzymatic pocket. It is envisaged that, by molecular linking the VHH to a cell penetrating peptide (CPP), the CPP-VHH fusion protein would be able to traverse the hydrophobic cell membrane into the cytoplasm and inhibit the intracellular BoNT. This presents a novel and safe immunotherapeutic strategy for botulism by using a cell penetrating, humanized-single domain antibody that inhibits the BoNT by means of a direct blockade of the groove of the menace enzyme.<\/jats:p>","DOI":"10.3390\/toxins3050469","type":"journal-article","created":{"date-parts":[[2011,5,13]],"date-time":"2011-05-13T11:42:42Z","timestamp":1305286962000},"page":"469-488","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":27,"title":["Botulinum Neurotoxins and Botulism: A Novel Therapeutic Approach"],"prefix":"10.3390","volume":"3","author":[{"given":"Jeeraphong","family":"Thanongsaksrikul","sequence":"first","affiliation":[{"name":"Laboratory for Research and Technology Development, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, 2 Prannok Road, Bangkok-noi, Bangkok 10700, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wanpen","family":"Chaicumpa","sequence":"additional","affiliation":[{"name":"Laboratory for Research and Technology Development, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, 2 Prannok Road, Bangkok-noi, Bangkok 10700, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2011,5,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1128\/mr.44.3.419-448.1980","article-title":"Clostridium botulinum neurotoxin","volume":"44","author":"Sugiyama","year":"1980","journal-title":"Microbiol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1128\/mr.46.1.86-94.1982","article-title":"Bacterial toxins: A table of lethal amounts","volume":"46","author":"Gill","year":"1982","journal-title":"Microbiol. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1128\/CMR.3.1.66","article-title":"Toxigenic clostridia","volume":"3","author":"Hatheway","year":"1990","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"225","DOI":"10.3201\/eid0802.010164","article-title":"Public health assessment of potential biological terrorism agents","volume":"8","author":"Rotz","year":"2002","journal-title":"Emerg. Infect. Dis."},{"key":"ref_5","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_6","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1016\/j.toxicon.2008.11.014","article-title":"A novel function of botulinum toxin-associated proteins: HA proteins disrupt intestinal epithelial barrier to increase toxin absorption","volume":"54","author":"Fuginaga","year":"2009","journal-title":"Toxicon"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1504\/TBJ.2008.018951","article-title":"How to botulinum neurotoxins block neurotransmitter release: From botulism to the molecular mechanism of action","volume":"1","author":"Poulain","year":"2008","journal-title":"The Botulinum J."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2345","DOI":"10.1128\/aem.58.8.2345-2354.1992","article-title":"Molecular cloning of the Clostridium botulinum structural gene encoding the type B neurotoxin and determination of its entire nucleotide sequence","volume":"58","author":"Whelan","year":"1992","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1082","DOI":"10.1101\/gr.6282807","article-title":"Genome sequence of a proteolytic (Group I) Clostridium botulinum strain Hall A and comparative analysis of the clostridial genomes","volume":"17","author":"Sebaihia","year":"2007","journal-title":"Genome Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1111\/j.1348-0421.1970.tb00495.x","article-title":"Conversion of toxigenicity in Clostridium botulinum type C","volume":"14","author":"Inoue","year":"1970","journal-title":"Jpn. J. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1038\/newbio235016a0","article-title":"Bacteriophage and the toxigenicity of Clostridium botulinum type D","volume":"235","author":"Eklund","year":"1972","journal-title":"Nat. New Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.1128\/aem.54.6.1405-1408.1988","article-title":"Evidence for plasmid-mediated toxin and bacteriocin production in Clostridium botulinum type G","volume":"54","author":"Eklund","year":"1988","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e4829","DOI":"10.1371\/journal.pone.0004829","article-title":"Evidence that plasmid-borne botulinum neurotoxin type B genes are widespread among Clostridium botulinum serotype B strains","volume":"4","author":"Franciosa","year":"2009","journal-title":"PLoS ONE"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"898","DOI":"10.1038\/2338","article-title":"Crystal structure of botulinum neurotoxin type A and implications for toxicity","volume":"5","author":"Lacy","year":"1998","journal-title":"Nat. Struct. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1038\/78005","article-title":"Structural analysis of the catalytic and binding sites of Clostridium botulinum neurotoxin B","volume":"7","author":"Swaminathan","year":"2000","journal-title":"Nat. Struct. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"23479","DOI":"10.1016\/S0021-9258(18)35863-0","article-title":"Botulinum neurotoxins are zinc protein","volume":"267","author":"Schiavo","year":"1992","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1016\/j.toxicon.2008.12.027","article-title":"Receptor and substrate interactions of clostridial neurotoxins","volume":"54","author":"Brunger","year":"2009","journal-title":"Toxicon"},{"key":"ref_18","unstructured":"Thanongsaksrikul, J. (2010). Therapeutic Antibodies for Disease Caused by Biological Weapon(s). Ph.D. Thesis, Mahidol University, Thailand."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"21950","DOI":"10.1074\/jbc.273.34.21950","article-title":"Binding and transcytosis of botulinum neurotoxin by polarized human colon carcinoma cells","volume":"273","author":"Maksymowych","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1073\/pnas.0609713104","article-title":"Identification of the protein receptor binding site of botulinum neurotoxins B and G proves the double-receptor concept","volume":"104","author":"Rummel","year":"2007","journal-title":"Proc. Natl. Acad. Sci.USA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1511","DOI":"10.1083\/jcb.200707184","article-title":"Mechanism of botulinum neurotoxin B and G entry into hippocampal neurons","volume":"179","author":"Dong","year":"2007","journal-title":"J. Cell. Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"592","DOI":"10.1126\/science.1123654","article-title":"SV2 is the protein receptor for botulinum neurotoxin A","volume":"312","author":"Dong","year":"2006","journal-title":"Science"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1038\/364827a0","article-title":"Direct visualization of botulinum neurotoxin-induced channels in phospholipid vesicles","volume":"364","author":"Schmidt","year":"1993","journal-title":"Nature"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1038\/nsb879","article-title":"Translocation of botulinum neurotoxin light chain protease through the heavy chain channel","volume":"10","author":"Koriazova","year":"2003","journal-title":"Nat. Struct. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2296","DOI":"10.1007\/s00018-008-8088-0","article-title":"Highly specific interaction between botulinum neurotoxins and synaptic vesicle proteins","volume":"65","author":"Brunger","year":"2008","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1152\/physrev.2000.80.2.717","article-title":"Neurotoxins affecting neuroexocytosis","volume":"80","author":"Schiavo","year":"2000","journal-title":"Physiol. Rev."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1038\/365160a0","article-title":"Botulinum neurotoxin A selectively cleaves the synapticprotein SNAP-25","volume":"365","author":"Blasi","year":"1993","journal-title":"Nature"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"23784","DOI":"10.1016\/S0021-9258(20)80452-9","article-title":"Identification of the nerve terminal targets of botulinum neurotoxin serotypes A, D, and E","volume":"268","author":"Schiavo","year":"1993","journal-title":"J. Biol. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1617","DOI":"10.1016\/S0021-9258(17)42071-0","article-title":"Proteolysis of SNAP-25 by types E and A botulinal neurotoxins","volume":"269","author":"Binz","year":"1994","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"10566","DOI":"10.1074\/jbc.270.18.10566","article-title":"Botulinum neurotoxin type C cleaves a single Lys-Ala bond within the carboxyl-terminal region of syntaxins","volume":"270","author":"Schiavo","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4821","DOI":"10.1002\/j.1460-2075.1993.tb06171.x","article-title":"Botulinum neurotoxin C1 blocks neurotransmitter release by means of cleaving HPC-1\/syntaxin","volume":"12","author":"Blasi","year":"1993","journal-title":"EMBO J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"832","DOI":"10.1038\/359832a0","article-title":"Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin","volume":"359","author":"Schiavo","year":"1992","journal-title":"Nature"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"11516","DOI":"10.1016\/S0021-9258(19)50230-7","article-title":"Botulinum neurotoxin serotype F is a zinc endopeptidase specific for VAMP\/synaptobrevin","volume":"268","author":"Schiavo","year":"1993","journal-title":"J. Biol. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"12764","DOI":"10.1016\/S0021-9258(18)99941-2","article-title":"Cleavage of members of the synaptobrevin\/VAMP family by types D and F botulinal neurotoxins and tetanus toxin","volume":"269","author":"Yamasaki","year":"1994","journal-title":"J. Biol. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1006\/bbrc.1994.1526","article-title":"Botulinum neurotoxin type G proteolyzes the Ala81-Ala82 bond of Rat synaptobrevin 2","volume":"200","author":"Yamasaki","year":"1994","journal-title":"Biochem. Biophys. Res."},{"key":"ref_36","unstructured":"Emmeluth, D. (2005). Botulism (Deadly Disease and Epidemics), Chelsea House Publishers. [1st]."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1111\/j.1432-1033.1994.tb19926.x","article-title":"Antagonism of the intracellular action of botulinum neurotoxin type A with monoclonal antibodies that map to light-chain epitopes","volume":"219","author":"Poulain","year":"1994","journal-title":"Eur. J. Biochem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"4648","DOI":"10.1021\/jm0300680","article-title":"Development of potent inhibitors of botulinum neurotoxin type B","volume":"46","author":"Anne","year":"2003","journal-title":"J. Med. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3055","DOI":"10.1016\/j.bmc.2004.03.006","article-title":"Small tripeptide surrogates with low nanomolar affinity as potent inhibitors of botulinum neurotoxin B metallo-proteolytic activity","volume":"12","author":"Blommaert","year":"2004","journal-title":"Bioorg. Med. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"643","DOI":"10.7326\/0003-4819-70-3-643","article-title":"Trivalent botulinus antotoxin","volume":"70","author":"Koenig","year":"1969","journal-title":"Ann. Intern. Med."},{"key":"ref_41","unstructured":"(2010). Investigational heptavalent botulinum antitoxin (HBAT) to replace licensed botulinum antitoxin AB and investigational botulinum antitoxin E. MMWR, 59, 299."},{"key":"ref_42","unstructured":"Available online:http:\/\/www.fda.gov\/downloads\/BiologicsBloodVaccines\/UCM149970.pdf."},{"key":"ref_43","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_44","doi-asserted-by":"crossref","first-page":"785","DOI":"10.1542\/peds.2006-0646","article-title":"Creation and development of the public service orphan drug human botulism immune globulin","volume":"119","author":"Arnon","year":"2007","journal-title":"Pediatrics"},{"key":"ref_45","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_46","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1016\/j.jmb.2006.09.084","article-title":"Fine and Domain-level epitope mapping of botulinum neurotoxin type A neutralizing antibodies by yeast surface display","volume":"365","author":"Levy","year":"2007","journal-title":"J. Mol. Biol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1093\/protein\/gzq001","article-title":"Affinity maturation of human botulinum neurotoxin antibodies by light chain shuffling via yeast mating","volume":"23","author":"Lou","year":"2010","journal-title":"Protein Eng. Des. Sel."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1016\/j.toxicon.2009.02.034","article-title":"Immune recognition of BoNTs A and B: how anti-toxin antibodies that bind to the heavy chain obstruct toxin action","volume":"54","author":"Atassi","year":"2009","journal-title":"Toxicon"},{"key":"ref_49","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_50","doi-asserted-by":"crossref","first-page":"602","DOI":"10.1128\/CMR.13.4.602","article-title":"Passive immunity in prevention of infectious diseases","volume":"13","author":"Keller","year":"2000","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1136\/bmj.1.5338.1123","article-title":"Human antitetanus serum in the treatment of tetanus","volume":"1","author":"Ellis","year":"1963","journal-title":"Br. Med. J."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"595","DOI":"10.2105\/AJPH.25.5.595","article-title":"Use of convalescent measles serum to control measles in a preparatory school","volume":"25","author":"Gallagher","year":"1935","journal-title":"Am. J. Public Health Nations Health"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"6191","DOI":"10.1128\/IAI.72.11.6191-6196.2004","article-title":"New concepts in antibody-mediated immunity","volume":"72","author":"Casadevall","year":"2004","journal-title":"Infect. Immun."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1016\/j.jaci.2006.01.015","article-title":"Use of intravenous immunoglobulin in human disease: A review of evidence by members of the primary immunodeficiency committee of the American academy of allergy, asthma and immunology","volume":"117","author":"Orange","year":"2006","journal-title":"J. Allerg. Clin. Immunol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"e16059","DOI":"10.1371\/journal.pone.0016059","article-title":"A broadly Flavivirus cross-neutralizing monoclonal antibody that recognizes a novel epitope within fushion loop of E protein","volume":"6","author":"Deng","year":"2011","journal-title":"PLoS ONE"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1394","DOI":"10.1128\/iai.63.4.1394-1399.1995","article-title":"Urease-specific monoclonal antibodies prevent Helicobacter felis infection in mice","volume":"63","author":"Blanchard","year":"1995","journal-title":"Infect. Immun."},{"key":"ref_57","first-page":"751","article-title":"Neutralization of measles virus infectivity and antibody-dependent cell-mediated cytotoxicity activity against an Epstein-Barr virus-infected cell line by intravenous administration of immunoglobulin G","volume":"10","author":"Colomar","year":"2003","journal-title":"Clin. Diagn. Lab. Immunol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1550","DOI":"10.4049\/jimmunol.167.3.1550","article-title":"Complement is essential for protection by an IgM and an IgG3 monoclonal antibody against experimental hematogenously disseminated candidiasis","volume":"167","author":"Han","year":"2001","journal-title":"J. Immunol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1038\/256495a0","article-title":"Continuous cultures of fused cells secreting antibody of predefined specificity","volume":"256","author":"Milstein","year":"1975","journal-title":"Nature"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"655","DOI":"10.1016\/j.drudis.2006.05.009","article-title":"Recombinant polyclonal antibodies: The next generation of antibody therapeutics?","volume":"11","author":"Haurum","year":"2006","journal-title":"Drug Discov. Today"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1093\/clinids\/21.1.150","article-title":"Return to the past: The case for antibody-based therapies in infectious diseases","volume":"21","author":"Casadevall","year":"1995","journal-title":"Clin. Infect. Dis."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1038\/nbt0409-331","article-title":"Development trends for therapeutic antibody fragments","volume":"27","author":"Nelson","year":"2009","journal-title":"Nat. Biotechnol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"1198","DOI":"10.1038\/aps.2010.120","article-title":"Research and development of next generation of antibody-based therapeutics","volume":"31","author":"Li","year":"2010","journal-title":"Acta. Pharmacol. Sin."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"e12416","DOI":"10.1371\/journal.pone.0012416","article-title":"Characterization of botulinum neurotoxin type A neutralizing monoclonal antibodies and influence of their half-lives on therapeutic activity","volume":"5","author":"Mazuet","year":"2010","journal-title":"PLoS ONE"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"2197","DOI":"10.1073\/pnas.77.4.2197","article-title":"Immunoglobulin chain loss in hybridoma lines","volume":"77","year":"1980","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_66","first-page":"4520","article-title":"Preexisting human anti-murine immunoglobulin reactivity due to polyclonal rheumatoid factors","volume":"47","author":"Epenetos","year":"1987","journal-title":"Cancer Res."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1038\/icb.1990.50","article-title":"Development of human anti-murine antibody (HAMA) response in patients","volume":"68","author":"Tjandra","year":"1990","journal-title":"Immunol. Cell Biol."},{"key":"ref_68","first-page":"1055","article-title":"Human immune response to anti-carcinoembryonic antigen murine monoclonal antibodies","volume":"1","author":"Losman","year":"1990","journal-title":"Cancer Res."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"6851","DOI":"10.1073\/pnas.81.21.6851","article-title":"Chimeric human antibody molecules: Mouse antigen-binding domains with human constant region domains","volume":"81","author":"Morrison","year":"1984","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.critrevonc.2004.10.011","article-title":"Overview of monoclonal antibodies in cancer therapy: Present and promise","volume":"54","author":"Stern","year":"2005","journal-title":"Crit. Rev. Oncol. Hematol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"673","DOI":"10.4049\/jimmunol.104.3.673","article-title":"Why antibody inhibits vascular clearance of staphylococcal enterotoxin B","volume":"104","author":"Normann","year":"1970","journal-title":"J. Immunol."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1046\/j.1432-1327.2001.01923.x","article-title":"Construction and functional evaluation of a single-chain antibody fragment that neutralizes toxin AahI from the venom of the scorpion Androctonus australis hector","volume":"268","author":"Devaux","year":"2001","journal-title":"Eur. J. Biochem."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1007\/s00424-009-0685-8","article-title":"Receptor-mediated endocytosis in renal proximal tubule","volume":"458","author":"Christensen","year":"2009","journal-title":"Pflug. Arch. Eur. J. Phys."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1038\/332323a0","article-title":"Reshaping human antibodies for therapy","volume":"332","author":"Riechmann","year":"1988","journal-title":"Nature"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1093\/protein\/gzp008","article-title":"Humanized-monoclonal antibody against heterologous Leptospira infection","volume":"22","author":"Maneewatch","year":"2009","journal-title":"Protein Eng. Des. Sel."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1038\/nrd1386","article-title":"Monoclonal antibodies market","volume":"3","author":"Reichert","year":"2004","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1016\/S0958-1669(97)80068-7","article-title":"Production of human antibody repertoires in transgenic mice","volume":"8","author":"Taussig","year":"1997","journal-title":"Curr. Opin. Biotechnol."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1084\/jem.156.3.930","article-title":"Use of Epstein-Barr virus-transformed B cell lines for the generation of immunoglobulin-producing human B cell hybridomas","volume":"156","author":"Chiorazzi","year":"1982","journal-title":"J. Exp. Med."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"2899","DOI":"10.4049\/jimmunol.130.6.2899","article-title":"Human monoclonal antibodies to a genus-specific chlamydial antigen produced by EBV-transformed B cells","volume":"130","author":"Rosen","year":"1983","journal-title":"J. Immunol."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"584","DOI":"10.1002\/eji.1830160522","article-title":"Human monoclonal antibody to purified protein derivative of tuberculin produced by hybrids constructed with Epstein-Barr virus-transformed B lymphocytes and mouse myeloma cells","volume":"16","author":"Garzelli","year":"1986","journal-title":"Eur. J. Immunol."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/0022-1759(84)90392-2","article-title":"Rescue of human monoclonal antibody production from an EBV-transformed B cell line by fusion to a human-mouse hybridoma","volume":"70","author":"Foung","year":"1984","journal-title":"J. Immunol. Methods"},{"key":"ref_82","unstructured":"Goldsby, R.A., Kindt, T.J., and Osborne, B.A. (2003). Kuby Immunology, W.H. Freeman and Company. [5th]. Chapter 5."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1146\/annurev.genet.39.073003.110544","article-title":"Immunoglobulin gene diversification","volume":"39","author":"Maizels","year":"2005","journal-title":"Annu. Rev. Genet."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1038\/nbt0697-553","article-title":"Yeast surface display for screening combinatorial polypeptide libraries","volume":"15","author":"Boder","year":"1997","journal-title":"Nat. Biotechnol."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1093\/protein\/12.7.613","article-title":"Development of an optimized expression system for the screening of antibody libraries displayed on the Escherichia coli surface","volume":"12","author":"Daugherty","year":"1999","journal-title":"Protein Eng."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"14336","DOI":"10.1073\/pnas.0805942105","article-title":"Isolation of human monoclonal antibodies by mammalian cell display","volume":"105","author":"Beerli","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"4937","DOI":"10.1073\/pnas.94.10.4937","article-title":"In vitro selection and evolution of functional proteins by using ribosome display","volume":"94","author":"Hanes","year":"1997","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"1315","DOI":"10.1126\/science.4001944","article-title":"Filamentous fusion phage: Novel expression vectors that display cloned antigens on the virion surface","volume":"228","author":"Smith","year":"1985","journal-title":"Science"},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"1105","DOI":"10.1038\/nbt1126","article-title":"Selecting and screening recombinant antibody libraries","volume":"23","author":"Hoogenboom","year":"2005","journal-title":"Nat. Biotechnol."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1016\/j.jprot.2008.12.007","article-title":"Human monoclonal ScFv neutralize lethal Thai cobra, Naja kaouthia, neurotoxin","volume":"72","author":"Kulkeaw","year":"2009","journal-title":"J. Proteomics"},{"key":"ref_91","doi-asserted-by":"crossref","unstructured":"O\u2019Brien, P.M., and Aitken, R. (2002). Antibody Phage Display: Methods and Protocols, Humana Press Inc.","DOI":"10.1385\/1592592406"},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"682","DOI":"10.1038\/nchembio843","article-title":"Synthetic therapeutic antibodies","volume":"2","author":"Sidhu","year":"2006","journal-title":"Nat. Chem. Biol."},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1146\/annurev.iy.12.040194.002245","article-title":"Making antibodies by phage display technology","volume":"12","author":"Winter","year":"1994","journal-title":"Annu. Rev. Immunol."},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"1267","DOI":"10.1093\/protein\/11.12.1267","article-title":"Absolute conservation of residue 6 of immunoglobulin heavy chain regions of class IIA is required for correct folding","volume":"11","author":"Kazemier","year":"1998","journal-title":"Protein Eng."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1177\/135965350901400204","article-title":"Human single-chain antibodies that neutralize homologous and heterologous strains and clades of influenza A virus subtype H5N1","volume":"14","author":"Maneewatch","year":"2009","journal-title":"Antivir. Ther."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.jviromet.2009.03.010","article-title":"Human single chain monoclonal antibody that recognizes matrix protein of heterologous influenza A virus subtype","volume":"159","author":"Poungpair","year":"2009","journal-title":"J. Virol. Methods"},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"9657","DOI":"10.1074\/jbc.M109.073163","article-title":"A VHH that neutralizes the zinc metalloproteinase activity of botulinum neurotoxin type A","volume":"285","author":"Thanongsaksrikul","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1475-2859-2-1","article-title":"The production of antibody fragments and antibody fusion proteins by yeasts and filamentous fungi","volume":"2","author":"Joosten","year":"2003","journal-title":"Microb. Cell. Fact."},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1038\/363446a0","article-title":"Naturally occurring antibodies devoid of light chains","volume":"363","author":"Atarhouch","year":"1993","journal-title":"Nature"},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/0014-5793(94)80432-X","article-title":"Camelising human antibody fragments: NMR studies on VH domains","volume":"339","author":"Davies","year":"1994","journal-title":"FEBS Lett."},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"1129","DOI":"10.1093\/protein\/7.9.1129","article-title":"Sequence and structure of VH domain from naturally occurring camel heavy chain immunoglobulins lacking light chains","volume":"7","author":"Muyldermans","year":"1994","journal-title":"Protein Eng."},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1093\/emboj\/19.5.921","article-title":"Camel heavy-chain antibodies: Diverse germline V(H)H and specific mechanisms enlarge the antigen-binding repertoire","volume":"19","author":"Nguyen","year":"2000","journal-title":"EMBO J."},{"key":"ref_103","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/S0145-305X(02)00071-X","article-title":"Emergence and evolution of functional heavy-chain antibodies in Camelidae","volume":"27","author":"Conrath","year":"2003","journal-title":"Dev. Comp. Immunol."},{"key":"ref_104","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1002\/prot.340160102","article-title":"Length distribution of CDRH3 in antibodies","volume":"16","author":"Wu","year":"1993","journal-title":"Proteins"},{"key":"ref_105","first-page":"277","article-title":"Single domain camel antibodies: Current status","volume":"74","author":"Muyldermans","year":"2001","journal-title":"J. Biotechnol."},{"key":"ref_106","doi-asserted-by":"crossref","first-page":"26285","DOI":"10.1074\/jbc.M102107200","article-title":"Antigen specificity and high affinity binding provided by one single loop of a camel single-domain antibody","volume":"276","author":"Desmyter","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_107","doi-asserted-by":"crossref","first-page":"4586","DOI":"10.1073\/pnas.0505379103","article-title":"Molecular basis for the preferential cleft recognition by dromedary heavy-chain antibodies","volume":"103","author":"Silence","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_108","doi-asserted-by":"crossref","first-page":"3512","DOI":"10.1093\/emboj\/17.13.3512","article-title":"Potent enzyme inhibitors derived from dromedary heavy-chain antibodies","volume":"17","author":"Lauwereys","year":"1998","journal-title":"EMBO J."},{"key":"ref_109","doi-asserted-by":"crossref","first-page":"2807","DOI":"10.1128\/AAC.45.10.2807-2812.2001","article-title":"\u03b2-lactamase inhibitors derived from single-domain antibody fragments elicited in the Camelidae","volume":"45","author":"Conrath","year":"2001","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_110","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/S0014-5793(97)01062-4","article-title":"Selection and identification of single domain antibody fragments from camel heavy-chain antibodies","volume":"414","author":"Ghahroudi","year":"1997","journal-title":"FEBS Lett."},{"key":"ref_111","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1007\/s00253-007-1142-2","article-title":"Properties, production, and applications of camelid single-domain antibody fragments","volume":"77","author":"Harmsen","year":"2007","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_112","unstructured":"Chulanetra, M., and Chaicumpa, W. Human antibody that neutralizes tetrodotoxin, Mahidol University, Thailand. Unpublished work, 2011."},{"key":"ref_113","first-page":"230","article-title":"Recognition of antigens by single domain antibody fragments: The superfluous luxury of paired domains","volume":"26","author":"Muyldermans","year":"2001","journal-title":"TIBS"},{"key":"ref_114","doi-asserted-by":"crossref","first-page":"1106","DOI":"10.1016\/j.jmb.2010.01.070","article-title":"A single-domain llama antibody potently inhibits the enzymatic activity of botulinum neurotoxin by binding to the non-catalytic \u03b1-exosite binding region","volume":"397","author":"Dong","year":"2010","journal-title":"J. Mol. Biol."},{"key":"ref_115","doi-asserted-by":"crossref","first-page":"e8818","DOI":"10.1371\/journal.pone.0008818","article-title":"Llama single domain antibodies specific for the 7 botulinum neurotoxin serotypes as heptaplex immunoreagents","volume":"5","author":"Conway","year":"2010","journal-title":"PLoS ONE"},{"key":"ref_116","doi-asserted-by":"crossref","first-page":"10444","DOI":"10.1016\/S0021-9258(17)34080-2","article-title":"The third helix of the Antennapedia homeodomain translocates through biological membranes","volume":"269","author":"Derossi","year":"1994","journal-title":"J. Biol. Chem."},{"key":"ref_117","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1096\/fasebj.12.1.67","article-title":"Cell penetration by transpotan","volume":"12","author":"Pooga","year":"1998","journal-title":"FASEB J."},{"key":"ref_118","doi-asserted-by":"crossref","first-page":"1134","DOI":"10.1021\/bc900251u","article-title":"A human single chain transbody specific to matrix protein (M1) interferes with the replication of Influenza A virus","volume":"21","author":"Poungpair","year":"2010","journal-title":"Bioconjugate Chem."},{"key":"ref_119","first-page":"1","article-title":"Protein transduction technology: A novel therapeutic perspective","volume":"60","author":"Noguchi","year":"2006","journal-title":"Acta Med. Okayama"},{"key":"ref_120","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1038\/90741","article-title":"Is VP22 nuclear homing an artifact?","volume":"19","author":"Lundberg","year":"2001","journal-title":"Nat. Biotechnol."},{"key":"ref_121","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1002\/jmr.637","article-title":"Membrane translocation of penetratin and its derivatives in different cell lines","volume":"16","author":"Letoha","year":"2003","journal-title":"J. Mol. Recognit."},{"key":"ref_122","doi-asserted-by":"crossref","first-page":"1246","DOI":"10.1021\/bc049938y","article-title":"Protein cargo delivery properties of cell-penetrating peptides: A comparative study","volume":"15","author":"Saalik","year":"2004","journal-title":"Bioconjugate Chem."},{"key":"ref_123","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/j.jconrel.2004.10.018","article-title":"Cationic TAT peptide transduction domain enters cells by macropinocytosis","volume":"102","author":"Kaplan","year":"2005","journal-title":"J. Control. Release"},{"key":"ref_124","doi-asserted-by":"crossref","first-page":"15300","DOI":"10.1074\/jbc.M401604200","article-title":"Cellular uptake of unconjugated TAT peptide involves clathrin-dependent endocytosis and heparin sulfate receptors","volume":"280","author":"Richard","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_125","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1042\/BC20070116","article-title":"Cell-penetrating peptides: From molecular mechanisms to therapeutics","volume":"100","author":"Morris","year":"2008","journal-title":"Biol. Cell"},{"key":"ref_126","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1002\/wnan.121","article-title":"Peptide-mediated cancer targeting of nanoconjugates","volume":"3","author":"Raha","year":"2010","journal-title":"WIREs Nanomed. Nanobiotechnol."},{"key":"ref_127","doi-asserted-by":"crossref","first-page":"1839","DOI":"10.1007\/s00018-005-5109-0","article-title":"Cell-penetrating peptides: Tools for intracellular delivery of therapeutics","volume":"62","author":"Deshayes","year":"2005","journal-title":"Cell Mol. Life Sci."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/3\/5\/469\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:56:05Z","timestamp":1760219765000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/3\/5\/469"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,5,13]]},"references-count":127,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2011,5]]}},"alternative-id":["toxins3050469"],"URL":"https:\/\/doi.org\/10.3390\/toxins3050469","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2011,5,13]]}}}