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Innovative and Highly Sensitive Detection of Clostridium perfringens Enterotoxin Based on Receptor Interaction and Monoclonal Antibodies
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,...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8068247/ https://www.ncbi.nlm.nih.gov/pubmed/33917845 http://dx.doi.org/10.3390/toxins13040266 |
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author | Neumann, Thea Krüger, Maren Weisemann, Jasmin Mahrhold, Stefan Stern, Daniel Dorner, Martin B. Feraudet-Tarisse, Cécile Pöhlmann, Christopher Schulz, Katharina Messelhäußer, Ute Rimek, Dagmar Gessler, Frank Elßner, Thomas Simon, Stéphanie Rummel, Andreas Dorner, Brigitte G. |
author_facet | Neumann, Thea Krüger, Maren Weisemann, Jasmin Mahrhold, Stefan Stern, Daniel Dorner, Martin B. Feraudet-Tarisse, Cécile Pöhlmann, Christopher Schulz, Katharina Messelhäußer, Ute Rimek, Dagmar Gessler, Frank Elßner, Thomas Simon, Stéphanie Rummel, Andreas Dorner, Brigitte G. |
author_sort | Neumann, Thea |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8068247 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-80682472021-04-25 Innovative and Highly Sensitive Detection of Clostridium perfringens Enterotoxin Based on Receptor Interaction and Monoclonal Antibodies Neumann, Thea Krüger, Maren Weisemann, Jasmin Mahrhold, Stefan Stern, Daniel Dorner, Martin B. Feraudet-Tarisse, Cécile Pöhlmann, Christopher Schulz, Katharina Messelhäußer, Ute Rimek, Dagmar Gessler, Frank Elßner, Thomas Simon, Stéphanie Rummel, Andreas Dorner, Brigitte G. Toxins (Basel) Article 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. MDPI 2021-04-08 /pmc/articles/PMC8068247/ /pubmed/33917845 http://dx.doi.org/10.3390/toxins13040266 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Neumann, Thea Krüger, Maren Weisemann, Jasmin Mahrhold, Stefan Stern, Daniel Dorner, Martin B. Feraudet-Tarisse, Cécile Pöhlmann, Christopher Schulz, Katharina Messelhäußer, Ute Rimek, Dagmar Gessler, Frank Elßner, Thomas Simon, Stéphanie Rummel, Andreas Dorner, Brigitte G. Innovative and Highly Sensitive Detection of Clostridium perfringens Enterotoxin Based on Receptor Interaction and Monoclonal Antibodies |
title | Innovative and Highly Sensitive Detection of Clostridium perfringens Enterotoxin Based on Receptor Interaction and Monoclonal Antibodies |
title_full | Innovative and Highly Sensitive Detection of Clostridium perfringens Enterotoxin Based on Receptor Interaction and Monoclonal Antibodies |
title_fullStr | Innovative and Highly Sensitive Detection of Clostridium perfringens Enterotoxin Based on Receptor Interaction and Monoclonal Antibodies |
title_full_unstemmed | Innovative and Highly Sensitive Detection of Clostridium perfringens Enterotoxin Based on Receptor Interaction and Monoclonal Antibodies |
title_short | Innovative and Highly Sensitive Detection of Clostridium perfringens Enterotoxin Based on Receptor Interaction and Monoclonal Antibodies |
title_sort | innovative and highly sensitive detection of clostridium perfringens enterotoxin based on receptor interaction and monoclonal antibodies |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8068247/ https://www.ncbi.nlm.nih.gov/pubmed/33917845 http://dx.doi.org/10.3390/toxins13040266 |
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