title: Development of Germline-Humanized Antibodies Neutralizing Botulinum Neurotoxin A and B.
abstract: Botulinum neurotoxins (BoNTs) are counted among the most toxic substances known and are responsible for human botulism, a life-threatening disease characterized by flaccid muscle paralysis that occurs naturally by food poisoning or colonization of the gastrointestinal tract by BoNT-producing clostridia. To date, 7 serologically distinct serotypes of BoNT (serotype A-G) are known. Due to the high toxicity of BoNTs the Centers for Disease Control and Prevention (CDC) have classified BoNTs as category A agent, including the six biological agents with the highest potential risk of use as bioweapons. Well tolerated antibodies neutralizing BoNTs are required to deal with the potential risk. In a previous work, we described the development of scFv and scFv-Fc (Yumab) from macaque origin (Macaca fascicularis) neutralizing BoNT/A and B by targeting the heavy and light chain of each serotype. In the present study, we humanized the macaque antibodies SEM120-IIIC1 (anti-BoNT/A light chain), A1HC38 (anti-BoNT/A heavy chain), BLC3 (anti-BoNT/B light chain) and B2-7 (anti-BoNT/B heavy chain) by germline-humanization to obtain a better potential immunotolerance in humans. We increased the Germinality Index (GI) of SEM120-IIIC1 to 94.5%, for A1HC38, to 95% for BLC3 and to 94.4% for B2-7. Furthermore, the neutralization efficacies of the germline-humanized antibodies were analyzed in lethal and non-lethal in vivo mouse assays as full IgG. The germline-humanized IgGs hu8SEM120-IIIC1, hu8A1HC38, hu8BLC3 and hu8B2-7 were protective in vivo, when anti-heavy and anti-light chain antibodies were combined. The synergistic effect and high humanness of the selected IgGs makes them promising lead candidates for further clinical development.
authors: Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie, Braunschweig, Germany.
authors: Sebastian
authors: S
authors: Miethe
authors: Institut Pasteur, Unité des Bactéries anaérobies et Toxines, Paris, France.
authors: Christelle
authors: C
authors: Mazuet
authors: National Institute for Biological Standards and Control (NIBSC), Division of Bacteriology, Potters Bar, United Kingdom.
authors: Yvonne
authors: Y
authors: Liu
authors: National Institute for Biological Standards and Control (NIBSC), Division of Bacteriology, Potters Bar, United Kingdom.
authors: Robert
authors: R
authors: Tierney
authors: National Institute for Biological Standards and Control (NIBSC), Division of Bacteriology, Potters Bar, United Kingdom.
authors: Christine
authors: C
authors: Rasetti-Escargueil
authors: Institut de Recherche Biomédicale des Armées (IRBA) Département des Maladies Infectieuses, Unité Interaction Hôte-Pathogène, Brétigny-sur-Orge, France.
authors: Arnaud
authors: A
authors: Avril
authors: Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie, Braunschweig, Germany.
authors: André
authors: A
authors: Frenzel
authors: Institut de Recherche Biomédicale des Armées (IRBA) Département des Maladies Infectieuses, Unité Interaction Hôte-Pathogène, Brétigny-sur-Orge, France.
authors: Philippe
authors: P
authors: Thullier
authors: Institut de Recherche Biomédicale des Armées (IRBA) Département des Maladies Infectieuses, Unité Interaction Hôte-Pathogène, Brétigny-sur-Orge, France.
authors: Thibaut
authors: T
authors: Pelat
authors: LFB Biotechnologies, Therapeutic Innovation Department, Lille, France.
authors: Remi
authors: R
authors: Urbain
authors: LFB Biotechnologies, Therapeutic Innovation Department, Lille, France.
authors: Alexandre
authors: A
authors: Fontayne
authors: National Institute for Biological Standards and Control (NIBSC), Division of Bacteriology, Potters Bar, United Kingdom.
authors: Dorothea
authors: D
authors: Sesardic
authors: Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie, Braunschweig, Germany.
authors: Michael
authors: M
authors: Hust
authors: Institut Pasteur, Unité des Bactéries anaérobies et Toxines, Paris, France.
authors: Michel Robert
authors: MR
authors: Popoff
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000276:immunology*; D057134:Antibodies, Neutralizing / Q000276:immunology; D019274:Botulinum Toxins, Type A / Q000276:immunology*; D001906:Botulism / Q000276:immunology; D003014:Clostridium botulinum; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D008252:Macaca fascicularis / Q000276:immunology; D051379:Mice; D009500:Neutralization Tests; D057127:Single-Chain Antibodies / Q000276:immunology
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D057134:Antibodies, Neutralizing; D007074:Immunoglobulin G; D057127:Single-Chain Antibodies; C096323:rimabotulinumtoxinB; D019274:Botulinum Toxins, Type A