Neutralization of Botulinum Neurotoxin Type E by a Humanized Antibody.
Botulinum neurotoxins (BoNTs) cause botulism and are the deadliest naturally-occurring substances known to humans. BoNTs have been classified as one of the category A agents by the Centers for Disease Control and Prevention, indicating their potential use as bioweapons. To counter bio-threat and naturally-occurring botulism cases, well-tolerated antibodies by humans that neutralize BoNTs are relevant. In our previous work, we showed the neutralizing potential of macaque (Macaca fascicularis)-derived scFv-Fc (scFv-Fc ELC18) by in vitro endopeptidase immunoassay and ex vivo mouse phrenic nerve-hemidiaphragm assay by targeting the light chain of the botulinum neurotoxin type E (BoNT/E). In the present study, we germline-humanized scFv-Fc ELC18 into a full IgG hu8ELC18 to increase its immunotolerance by humans. We demonstrated the protection and prophylaxis capacity of hu8ELC18 against BoNT/E in a mouse model. A concentration of 2.5 ng/mouse of hu8ELC18 protected against 5 mouse lethal dose (MLD) in a mouse protection assay and complete neutralization of 1 LD50 of pure BoNT/E toxin was achieved with 8 ng of hu8ELC18 in mouse paralysis assay. Furthermore, hu8ELC18 protected mice from 5 MLD if injected up to 14 days prior to intraperitoneal BoNT/E administration. This newly-developed humanized IgG is expected to have high tolerance in humans.
Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, University of Helsinki, Helsinki 00014, Finland. yagmur.derman@helsinki.fi.
Yağmur
Y
Derman
Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, University of Helsinki, Helsinki 00014, Finland. katja.selby@helsinki.fi.
Katja
K
Selby
Institut für Biochemie, Biotechnologie, und Bioinformatik, Technische Universität Braunschweig, Abteilung Biotechnologie, Braunschweig 38106, Germany. s.miethe@tu-braunschweig.de.
Sebastian
S
Miethe
Institut für Biochemie, Biotechnologie, und Bioinformatik, Technische Universität Braunschweig, Abteilung Biotechnologie, Braunschweig 38106, Germany. andre.frenzel@tu-bs.de.|YUMAB GmbH, Rebenring 33, Braunschweig 38106, Germany. andre.frenzel@tu-bs.de.
André
A
Frenzel
Division of Bacteriology, National Institute for Biological Standards and Control (NIBSC), A centre of Medicine and Healthcare products Regulatory Agency, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. yvonne.liu@nibsc.org.
Yvonne
Y
Liu
Division of Bacteriology, National Institute for Biological Standards and Control (NIBSC), A centre of Medicine and Healthcare products Regulatory Agency, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. christine.escargueil@laposte.net.
Christine
C
Rasetti-Escargueil
Institut de Recherche Biomédicale des Armées (IRBA), Département des Maladies Infectieuses, Unité biothérapies anti-infectieuses et immunité, Brétigny-sur-Orge, 1 place du Général Valérie André 91220, France. arnaud.avril@irba.fr.
Arnaud
A
Avril
Institut de Recherche Biomédicale des Armées (IRBA), Département des Maladies Infectieuses, Unité biothérapies anti-infectieuses et immunité, Brétigny-sur-Orge, 1 place du Général Valérie André 91220, France. thibaut.pelat@biotem.fr.
Thibaut
T
Pelat
LFB Biotechnologies, Therapeutic Innovation Department, 59 Rue de Trévise, Lille Cedex BP 62006-59011, France. remi.urbain@ecdysispharma.com.|Ecdysis Pharma, Bioincubateur Eurasanté, 70 rue du Dr Yersin, Loos 59120, France. remi.urbain@ecdysispharma.com.
Remi
R
Urbain
LFB Biotechnologies, Therapeutic Innovation Department, 59 Rue de Trévise, Lille Cedex BP 62006-59011, France. fontaynea@lfb.fr.
Alexandre
A
Fontayne
Institut de Recherche Biomédicale des Armées (IRBA), Département des Maladies Infectieuses, Unité biothérapies anti-infectieuses et immunité, Brétigny-sur-Orge, 1 place du Général Valérie André 91220, France. pthullier@yahoo.com.
Philippe
P
Thullier
Division of Bacteriology, National Institute for Biological Standards and Control (NIBSC), A centre of Medicine and Healthcare products Regulatory Agency, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. Thea.Sesardic@nibsc.org.
Dorothea
D
Sesardic
Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, University of Helsinki, Helsinki 00014, Finland. miia.lindstrom@helsinki.fi.
Miia
M
Lindström
Institut für Biochemie, Biotechnologie, und Bioinformatik, Technische Universität Braunschweig, Abteilung Biotechnologie, Braunschweig 38106, Germany. m.hust@tu-bs.de.
Michael
M
Hust
Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, University of Helsinki, Helsinki 00014, Finland. hannu.korkeala@helsinki.fi.
Hannu
H
Korkeala
D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000494:pharmacology*; D057134:Antibodies, Neutralizing / Q000494:pharmacology*; D000931:Antidotes / Q000494:pharmacology*; D000992:Antitoxins / Q000494:pharmacology*; D001905:Botulinum Toxins / Q000037:antagonists & inhibitors* / Q000276:immunology; D001906:Botulism / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D003014:Clostridium botulinum / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D004195:Disease Models, Animal; D005260:Female; D051379:Mice; D057127:Single-Chain Antibodies / Q000494:pharmacology*
D061067:Antibodies, Monoclonal, Humanized; D057134:Antibodies, Neutralizing; D000931:Antidotes; D000992:Antitoxins; D057127:Single-Chain Antibodies; D001905:Botulinum Toxins; C096326:botulinum toxin type E
antibody; botulinum neurotoxin type E; botulism