Development of neutralizing scFv-Fc against botulinum neurotoxin A light chain from a macaque immune library
| Autor: | Thomas Schirrmann, Yvonne Liu, Sebastian Miethe, André Frenzel, Christine Rasetti-Escargueil, Philippe Thullier, Arnaud Avril, Dorothea Sesardic, Michael Hust, Thibaut Pelat, Siham Chahboun |
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| Přispěvatelé: | Institut für Biochemie, Biotechnologie und Bioinformatik [TU, Braunschweig], Technische Universität Braunschweig = Technical University of Braunschweig [Braunschweig], Division of Bacteriology, National Institute for Biological Standards and Control (NIBSC), Medicines and Healthcare Products Regulatory Agency (MHRA)-Medicines and Healthcare Products Regulatory Agency (MHRA), Division of Bacteriology [UK], Unité de biotechnologie des anticorps et des toxines, Centre de Recherches du Service de Santé des Armées (CRSSA)-Institut de Recherche Biomédicale des Armées (IRBA), Centre de Recherches du Service de Santé des Armées (CRSSA), Service de Santé des Armées, We acknowledge funding from the European Community’s Seventh Framework Program (FP7/2007–2013) under agreement no. 241832 granted to the AntiBotABE project (http://www.antibotabe.com)., We would like to thank Nicola Bak for helping us with the ex-vivo assays., Technische Universität Braunschweig [Braunschweig] |
| Rok vydání: | 2014 |
| Předmět: |
MESH: Botulinum Toxins
Type A / immunology Male Phage display [SDV]Life Sciences [q-bio] medicine.disease_cause Neutralization scFv MESH: Paralysis / immunology Mice Endopeptidase activity MESH: Phrenic Nerve / drug effects BoNT/A Clostridium botulinum Immunology and Allergy MESH: Immunoglobulin Light Chains Surrogate / genetics Botulism MESH: Animals Botulinum Toxins Type A in vitro assay MESH: Immunoglobulin Fc Fragments / genetics Mice Inbred BALB C MESH: Botulism / immunology MESH: Antibodies Blocking / metabolism macaque MESH: Antibodies Blocking / genetics botulinum neurotoxin MESH: Botulism / complications ex-vivo assay 3. Good health Phrenic Nerve mouse phrenic nerve-hemidiaphragm assay [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology MESH: Cell Surface Display Techniques MESH: Immunization Immunotherapy Antibody phage display MESH: Paralysis / prevention & control medicine.drug_class MESH: Immunotherapy / methods Immunoglobulin Light Chains Surrogate Immunology MESH: Mice Inbred BALB C Biology Monoclonal antibody MESH: Paralysis / etiology MESH: Immunoglobulin Light Chains Surrogate / metabolism MESH: Botulism / therapy Report medicine Animals Humans Paralysis MESH: Phrenic Nerve / immunology Antibodies Blocking MESH: Mice MESH: Immunoglobulin Light Chains Surrogate / administration & dosage scFv-Fc MESH: Antibodies Blocking / administration & dosage MESH: Epitope Mapping MESH: Humans Toxin MESH: Single-Chain Antibodies / metabolism Clostridium botulinum type A Immunity medicine.disease MESH: Single-Chain Antibodies / genetics MESH: Botulinum Toxins Type A / adverse effects Virology [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology MESH: Male Immunoglobulin Fc Fragments Macaca fascicularis MESH: Immunoglobulin Fc Fragments / metabolism MESH: Macaca fascicularis biology.protein SNAP-25 Immunization MESH: Clostridium botulinum type A / immunology MESH: Immunity / genetics Cell Surface Display Techniques Epitope Mapping Single-Chain Antibodies |
| Zdroj: | mAbs mAbs, Taylor & Francis, 2014, 6 (2), pp.446-459. ⟨10.4161/mabs.27773⟩ |
| ISSN: | 1942-0870 1942-0862 |
| DOI: | 10.4161/mabs.27773⟩ |
| Popis: | International audience; Botulinum toxins (BoNTs) are among the most toxic substances on earth, with serotype A toxin being the most toxic substance known. They are responsible for human botulism, a disease characterized by flaccid muscle paralysis that occurs naturally through food poisoning or the colonization of the gastrointestinal tract by BoNT-producing clostridia. BoNT has been classified as a category A agent by the Centers for Disease Control, and it is one of six agents with the highest potential risk of use as bioweapons. Human or human-like neutralizing antibodies are thus required for the development of anti-botulinum toxin drugs to deal with this possibility. In this study, Macaca fascicularis was hyperimmunized with a recombinant light chain of BoNT/A. An immune phage display library was constructed and, after multistep panning, several scFv with nanomolar affinities that inhibited the endopeptidase activity of BoNT/A1 in vitro as scFv-Fc, with a molar ratio (ab binding site:toxin) of up to 1:1, were isolated. The neutralization of BoNT/A-induced paralysis by the SEM120-IID5, SEM120-IIIC1 and SEM120-IIIC4 antibodies was demonstrated in mouse phrenic nerve-hemidiaphragm preparations with the holotoxin. The neutralization observed is the strongest ever measured in the phrenic nerve-hemidiaphragm assay for BoNT/A1 for a monoclonal antibody. Several scFv-Fc inhibiting the endopeptidase activity of botulinum neurotoxin A were isolated. For SEM120-IID5, SEM120-IIIC1, and SEM120-IIIC4, inhibitory effects in vitro and protection against the toxin ex vivo were observed. The human-like nature of these antibodies makes them promising lead candidates for further development of immunotherapeutics for this disease. |
| Databáze: | OpenAIRE |
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