An optical biosensor assay for rapid dual detection of Botulinum neurotoxins A and E
| Autor: | Marie-Pierre Blanchard, Yves Maulet, Michael Seagar, Christelle Mazuet, Michel R. Popoff, Christian Lévêque, Oussama El Far, Géraldine Ferracci |
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| Přispěvatelé: | Unité de Neurobiologie des canaux Ioniques et de la Synapse (UNIS - Inserm U1072), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre de recherche en neurobiologie - neurophysiologie de Marseille (CRN2M), Centre National de Référence des Bactéries Anaérobies et Botulisme - National Reference Center Anaerobic Bacteria and Botulism (CNR), Institut Pasteur [Paris] (IP), This work was supported by INSERM, CNRS and a grant from the DGA (REI N° 0634040), El Far, Oussama, Institut Pasteur [Paris] |
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Botulinum Toxins [SDV.BIO]Life Sciences [q-bio]/Biotechnology [SDV]Life Sciences [q-bio] Biosensing Techniques 01 natural sciences Epitope Substrate Specificity Epitopes Lab-On-A-Chip Devices MESH: Microchip Analytical Procedures Botulinum Toxins Type A Surface plasmon resonance CHIP assay Multidisciplinary [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology MESH: Kinetics Chemistry Antibodies Monoclonal Optical biosensor MESH: Lab-On-A-Chip Devices [SDV.TOX] Life Sciences [q-bio]/Toxicology [SDV] Life Sciences [q-bio] [SDV.TOX]Life Sciences [q-bio]/Toxicology MESH: Botulinum Toxins Type A*/immunology [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] MESH: Biosensing Techniques MESH: Botulinum Toxins*/immunology MESH: Enzyme Activation medicine.drug_class Monoclonal antibody Sensitivity and Specificity Article 03 medical and health sciences In vivo Microchip Analytical Procedures MESH: Antibodies Monoclonal/immunology medicine Humans [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] MESH: Humans MESH: Epitopes/immunology 010401 analytical chemistry Serum samples Molecular biology MESH: Sensitivity and Specificity 0104 chemical sciences [SDV.BIO] Life Sciences [q-bio]/Biotechnology Enzyme Activation Kinetics 030104 developmental biology MESH: Substrate Specificity Biosensor [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology |
| Zdroj: | Scientific Reports Scientific Reports, 2015, 5 (1), pp.17953. ⟨10.1038/srep17953⟩ Scientific Reports, Nature Publishing Group, 2015, 5 (1), pp.17953. ⟨10.1038/srep17953⟩ |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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