Innovative and Highly Sensitive Detection of
| Autor: | Brigitte G. Dorner, Stefan Mahrhold, Daniel Stern, Ute Messelhäußer, Thea Neumann, Dagmar Rimek, Maren Krüger, Martin B. Dorner, Frank Gessler, Christopher Pöhlmann, Thomas Elßner, Katharina Schulz, Andreas Rummel, Cécile Feraudet-Tarisse, Jasmin Weisemann, Stéphanie Simon |
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| Přispěvatelé: | 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) |
| Rok vydání: | 2021 |
| Předmět: |
Clostridium perfringens
viruses Health Toxicology and Mutagenesis Antibody Affinity lcsh:Medicine mobile on-site detection Enterotoxin Toxicology medicine.disease_cause Epitope law.invention Workflow Foodborne Diseases Enterotoxins Epitopes Feces Mice C. perfringens enterotoxin CPE law Antibody Specificity Limit of Detection [SDV.IDA]Life Sciences [q-bio]/Food engineering stationary lab-based detection Claudin-4 Receptor 0303 health sciences Chemistry digestive oral and skin physiology Antibodies Monoclonal 3. Good health [SDV.TOX]Life Sciences [q-bio]/Toxicology receptor claudin-4 Recombinant DNA monoclonal antibodies Protein Binding medicine.drug_class Enzyme-Linked Immunosorbent Assay Monoclonal antibody Article 03 medical and health sciences stomatognathic system Predictive Value of Tests medicine Animals Humans ddc:610 030304 developmental biology Automation Laboratory urogenital system 030306 microbiology lcsh:R Reproducibility of Results biochemical phenomena metabolism and nutrition Molecular biology In vitro Highly sensitive Clostridium Infections 610 Medizin und Gesundheit Epitope Mapping |
| Zdroj: | Toxins Toxins, MDPI, 2021, 13 (4), ⟨10.3390/toxins13040266⟩ Volume 13 Issue 4 Toxins, 2021, 13 (4), ⟨10.3390/toxins13040266⟩ Toxins, Vol 13, Iss 266, p 266 (2021) |
| ISSN: | 2072-6651 |
| DOI: | 10.3390/toxins13040266⟩ |
| Popis: | Clostridium perfringens enterotoxin (CPE) regularly causes food poisoning and antibiotic-associated diarrhea therefore, reliable toxin detection is crucial. To this aim, we explored stationary and mobile strategies to detect CPE either exclusively by monoclonal antibodies (mAbs) or, alternatively, by toxin-enrichment via the cellular receptor of CPE, claudin-4, and mAb detection. Among the newly generated mAbs, we identified nine CPE-specific mAbs targeting five distinct epitopes, among them mAbs recognizing CPE bound to claudin-4 or neutralizing CPE activity in vitro. In surface plasmon resonance experiments, all mAbs and claudin-4 revealed excellent affinities towards CPE, ranging from 0.05 to 2.3 nM. Integrated into sandwich enzyme-linked immunosorbent assays (ELISAs), the most sensitive mAb/mAb and claudin-4/mAb combinations achieved similar detection limits of 0.3 pg/mL and 1.0 pg/mL, respectively, specifically detecting recombinant CPE from spiked feces and native CPE from 30 different C. perfringens culture supernatants. The implementation of mAb- and receptor-based ELISAs into a mobile detection platform enabled the fast detection of CPE, which will be helpful in clinical laboratories to diagnose diarrhea of assumed bacterial origin. In conclusion, we successfully employed an endogenous receptor and novel high affinity mAbs for highly sensitive and specific CPE-detection. These tools will be useful for both basic and applied research. |
| Databáze: | OpenAIRE |
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