Characterization of a highly neutralizing single monoclonal antibody to botulinum neurotoxin type A

Autor: Christine Rasetti-Escargueil, Anne Wijkhuisen, Emmanuel Lemichez, Stéphanie Simon, Michel R. Popoff, Sébastien Brier, Maud Marechal
Přispěvatelé: Plateforme technologique de RMN biologique - Biological NMR Technological Platform, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Toxines bactériennes - Bacterial Toxins, Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), French joint ministerial program against CBRNE, ANR-11-EQPX-0008,CACSICE,Centre d'analyse de systèmes complexes dans les environnements complexes(2011), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)
Rok vydání: 2021
Předmět:
0301 basic medicine
Gene isoform
medicine.drug_class
Protein Conformation
Nerve Tissue Proteins
Monoclonal antibody
Biochemistry
Epitope
Neutralization
03 medical and health sciences
Epitopes
Mice
0302 clinical medicine
Gangliosides
Genetics
medicine
Animals
Botulism
Botulinum Toxins
Type A
Molecular Biology
mass spectrometry
Antiserum
Membrane Glycoproteins
botulism
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Structural Biology [q-bio.BM]
Chemistry
Hydrogen-Deuterium eXchange
botulinum neurotoxin
synaptic vesicle protein 2
Antibodies
Monoclonal
medicine.disease
Molecular biology
Antibodies
Neutralizing
epitope mapping
030104 developmental biology
Epitope mapping
Neuromuscular Agents
GT1b
monoclonal antibody
030217 neurology & neurosurgery
Biotechnology
Conformational epitope
Zdroj: FASEB Journal
FASEB Journal, 2021, 35 (5), ⟨10.1096/fj.202002492R⟩
FASEB Journal, Federation of American Society of Experimental Biology, 2021, 35 (5), ⟨10.1096/fj.202002492R⟩
ISSN: 1530-6860
0892-6638
DOI: 10.1096/fj.202002492R⟩
Popis: International audience; Compared to conventional antisera strategies, monoclonal antibodies (mAbs) represent an alternative and safer way to treat botulism, a fatal flaccid paralysis due to botulinum neurotoxins (BoNTs). In addition, mAbs offer the advantage to be produced in a reproducible manner. We previously identified a unique and potent mouse mAb (TA12) targeting BoNT/A1 with high affinity and neutralizing activity. In this study, we characterized the molecular basis of TA12 neutralization by combining Hydrogen/Deuterium eXchange Mass Spectrometry (HDX-MS) with site-directed mutagenesis and functional studies. We found that TA12 recognizes a conformational epitope located at the interface between the HCN and HCC subdomains of the BoNT/A1 receptor-binding domain (HC ). The TA12-binding interface shares common structural features with the ciA-C2 VHH epitope and lies on the face opposite recognized by ciA-C2- and the CR1/CR2-neutralizing mAbs. The single substitution of N1006 was sufficient to affect TA12 binding to HC confirming the position of the epitope. We further uncovered that the TA12 epitope overlaps with the BoNT/A1-binding site for both the neuronal cell surface receptor synaptic vesicle glycoprotein 2 isoform C (SV2C) and the GT1b ganglioside. Hence, TA12 potently blocks the entry of BoNT/A1 into neurons by interfering simultaneously with the binding of SV2C and to a lower extent GT1b. Our study reveals the unique neutralization mechanism of TA12 and emphasizes on the potential of using single mAbs for the treatment of botulism type A.
Databáze: OpenAIRE
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