title: An optical biosensor assay for rapid dual detection of Botulinum neurotoxins A and E.
abstract: The enzymatic activity of the pathogenic botulinum neurotoxins type A and E (BoNT/A and E) leads to potentially lethal paralytic symptoms in humans and their prompt detection is of crucial importance. A chip assay based on Surface Plasmon Resonance monitoring of the cleavage products is a simple method that we have previously established to detect BoNT/A activity. We have now developed a similar format assay to measure BoNT/E activity. A monoclonal antibody specifically recognizing SNAP25 cleaved by BoNT/E was generated and used to measure the appearance of the neo-epitope following injection of BoNT/E over SNAP-25 immobilized on a chip. This assay detects BoNT/E activity at 1 LD50/ml within minutes and linear dose-responses curves were obtained using a multiplexed biosensor. A threshold of 0.01 LD50/ml was achieved after 5 h of cleavage. This assay is 10-fold more sensitive than the in vivo assay for direct detection of BoNT/E in serum samples. The SNAP25 chip assay is able to discriminate in an automated manner the presence of BoNT/E, BoNT/A or a combination of both toxins.
authors.affiliation: INSERM, UMR_S 1072, 13015 Marseille, France.|Aix-Marseille Université, 13015 Marseille, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Lévêque
authors.affiliation: CNRS, UMR 7286, Plate-Forme de Recherche en Neurosciences PFRN, 13015 Marseille, France.|Aix-Marseille Université, 13015 Marseille, France.
authors.forename: Géraldine
authors.initials: G
authors.lastname: Ferracci
authors.affiliation: INSERM, UMR_S 1072, 13015 Marseille, France.|Aix-Marseille Université, 13015 Marseille, France.
authors.forename: Yves
authors.initials: Y
authors.lastname: Maulet
authors.affiliation: CNR Bactéries Anaérobies et botulisme, Unité des Bactéries anaérobies et toxines. Institut Pasteur, 28 rue du Dr Roux, 75724 Paris Cedex 15, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Mazuet
authors.affiliation: CNR Bactéries Anaérobies et botulisme, Unité des Bactéries anaérobies et toxines. Institut Pasteur, 28 rue du Dr Roux, 75724 Paris Cedex 15, France.
authors.forename: Michel R
authors.initials: MR
authors.lastname: Popoff
authors.affiliation: CNRS, UMR 7286, Plate-Forme de Recherche en Neurosciences PFRN, 13015 Marseille, France.|Aix-Marseille Université, 13015 Marseille, France.
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Blanchard
authors.affiliation: INSERM, UMR_S 1072, 13015 Marseille, France.|Aix-Marseille Université, 13015 Marseille, France.
authors.forename: Michael
authors.initials: M
authors.lastname: Seagar
authors.affiliation: INSERM, UMR_S 1072, 13015 Marseille, France.|Aix-Marseille Université, 13015 Marseille, France.
authors.forename: Oussama
authors.initials: O
authors.lastname: El Far
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D015374:Biosensing Techniques; D001905:Botulinum Toxins / Q000276:immunology; D019274:Botulinum Toxins, Type A / Q000276:immunology; D004789:Enzyme Activation; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007700:Kinetics; D056656:Lab-On-A-Chip Devices; D046208:Microchip Analytical Procedures; D012680:Sensitivity and Specificity; D013379:Substrate Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A; C545476:incobotulinumtoxinA