Isolation of nanomolar scFvs of non-human primate origin, cross-neutralizing botulinum neurotoxins A1 and A2 by targeting their heavy chain

Autor: Siham Chahboun, Arnaud Avril, Michael Hust, Thibaut Pelat, Dorothea Sesardic, Christine Rasetti-Escargueil, Christelle Mazuet, Sebastian Miethe, Michel R. Popoff, Philippe Thullier
Přispěvatelé: Institut de Recherche Biomédicale des Armées [Brétigny-sur-Orge] (IRBA), Technische Universität Braunschweig = Technical University of Braunschweig [Braunschweig], Centre National de Référence des Bactéries Anaérobies et Botulisme - National Reference Center Anaerobic Bacteria and Botulism (CNR), Institut Pasteur [Paris] (IP), National Institute for Biological Standards and Control (NIBSC), Medicines and Healthcare Products Regulatory Agency (MHRA), BIOTEM, We acknowledge funding from the European Community Seventh Framework Program (FP7/2007–2013) under agreement N° 241832 granted to the AntiBotABE project (http://www.antibotabe.com)., We thank Olivier De Bardoneche (Absiskey) for his excellent project management administrative work on the AntibotABE project. We thank Jordi Molgo, Hannu Korkeala, Yagmur Derman, Katja Selby, Alexandre Fontayne and Audrey Merens for their active participation to the AntiBotABE consortium and for their scientific expertise. We thank Saskia Helmsing for technical assistance, European Project: 241832,EC:FP7:SEC,FP7-SEC-2009-1,ANTIBOTABE(2010), Institut de Recherche Biomédicale des Armées (IRBA), Institut Pasteur [Paris], Institut de Recherche Biomédicale des Armées ( IRBA ), Technische Universität Braunschweig [Braunschweig], Centre National de Référence des Bactéries Anaérobies et du Botulisme - Bactéries Anaérobies et Toxines ( CNR ), National Institute for Biological Standards and Control ( NIBSC ), European Project : 241832,EC:FP7:SEC,FP7-SEC-2009-1,ANTIBOTABE ( 2010 )
Rok vydání: 2015
Předmět:
Zdroj: BMC Biotechnology
BMC Biotechnology, 2015, 15 (1), pp.86. ⟨10.1186/s12896-015-0206-0⟩
BMC Biotechnology, BioMed Central, 2015, 15 (1), pp.86. ⟨10.1186/s12896-015-0206-0⟩
BMC Biotechnology, BioMed Central, 2015, 15 (1), pp.86. 〈10.1186/s12896-015-0206-0〉
ISSN: 1472-6750
Popis: Background Botulism is a naturally occurring disease, mainly caused by the ingestion of food contaminated by the botulinum neurotoxins (BoNTs). Botulinum neurotoxins are the most lethal. They are classified among the six major biological warfare agents by the Centers for Disease Control. BoNTs act on the cholinergic motoneurons, where they cleave proteins implicated in acetylcholine vesicle exocytosis. This exocytosis inhibition induces a flaccid paralysis progressively affecting all the muscles and generally engendering a respiratory distress. BoNTs are also utilized in medicine, mainly for the treatment of neuromuscular disorders, preventing large scale vaccination. Botulism specific treatment requires injections of antitoxins, usually of equine origin and thus poorly tolerated. Therefore, development of human or human-like neutralizing antibodies is of a major interest, and it is the subject of the European framework project called “AntiBotABE”. Results In this study, starting from a macaque immunized with the recombinant heavy chain of BoNT/A1 (BoNT/A1-HC), an immune antibody phage-display library was generated and antibody fragments (single chain Fragment variable) with nanomolar affinity were isolated and further characterized. The neutralization capacities of these scFvs were analyzed in the mouse phrenic nerve-hemidiaphragm assay. Conclusions After a three-round panning, 24 antibody fragments with affinity better than 10 nM were isolated. Three of them neutralized BoNT/A1 efficiently and two cross-neutralized BoNT/A1 and BoNT/A2 subtypes in the mouse phrenic nerve-hemidiaphragm assay. These are the first monoclonal human-like antibodies cross-neutralizing both BoNT/A1 and BoNT/A2. The antibody A1HC38 was selected for further development, and could be clinically developed for the prophylaxis and treatment of botulism. Electronic supplementary material The online version of this article (doi:10.1186/s12896-015-0206-0) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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