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A Monoclonal–Monoclonal Antibody Based Capture ELISA for Abrin

Abrin, one of the most highly potent toxins in the world, is derived from the plant, Abrus precatorius. Because of its high toxicity, it poses potential bioterror risks. Therefore, a need exists for new reagents and technologies that would be able to rapidly detect abrin contamination as well as lea...

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Autores principales: Tam, Christina C., Cheng, Luisa W., He, Xiaohua, Merrill, Paul, Hodge, David, Stanker, Larry H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5666375/
https://www.ncbi.nlm.nih.gov/pubmed/29057799
http://dx.doi.org/10.3390/toxins9100328
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author Tam, Christina C.
Cheng, Luisa W.
He, Xiaohua
Merrill, Paul
Hodge, David
Stanker, Larry H.
author_facet Tam, Christina C.
Cheng, Luisa W.
He, Xiaohua
Merrill, Paul
Hodge, David
Stanker, Larry H.
author_sort Tam, Christina C.
collection PubMed
description Abrin, one of the most highly potent toxins in the world, is derived from the plant, Abrus precatorius. Because of its high toxicity, it poses potential bioterror risks. Therefore, a need exists for new reagents and technologies that would be able to rapidly detect abrin contamination as well as lead to new therapeutics. We report here a group of abrin-specific monoclonal antibodies (mAbs) that recognize abrin A-chain, intact A–B chain toxin, and agglutinin by Western blot. Additionally, these mAbs were evaluated for their ability to serve as capture antibodies for a sandwich (capture) ELISA. All possible capture–detector pairs were evaluated and the best antibody pair identified and optimized for a capture ELISA. The capture ELISA based on this capture–detector mAb pair had a limit of detection (L.O.D) of ≈1 ng/mL measured using three independent experiments. The assay did not reveal any false positives with extracts containing other potential ribosome-inactivating proteins (RIPs). Thus, this new capture ELISA uses mAbs for both capture and detection; has no cross-reactivity against other plant RIPs; and has a sensitivity comparable to other reported capture ELISAs using polyclonal antibodies as either capture or detector.
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spelling pubmed-56663752017-11-09 A Monoclonal–Monoclonal Antibody Based Capture ELISA for Abrin Tam, Christina C. Cheng, Luisa W. He, Xiaohua Merrill, Paul Hodge, David Stanker, Larry H. Toxins (Basel) Article Abrin, one of the most highly potent toxins in the world, is derived from the plant, Abrus precatorius. Because of its high toxicity, it poses potential bioterror risks. Therefore, a need exists for new reagents and technologies that would be able to rapidly detect abrin contamination as well as lead to new therapeutics. We report here a group of abrin-specific monoclonal antibodies (mAbs) that recognize abrin A-chain, intact A–B chain toxin, and agglutinin by Western blot. Additionally, these mAbs were evaluated for their ability to serve as capture antibodies for a sandwich (capture) ELISA. All possible capture–detector pairs were evaluated and the best antibody pair identified and optimized for a capture ELISA. The capture ELISA based on this capture–detector mAb pair had a limit of detection (L.O.D) of ≈1 ng/mL measured using three independent experiments. The assay did not reveal any false positives with extracts containing other potential ribosome-inactivating proteins (RIPs). Thus, this new capture ELISA uses mAbs for both capture and detection; has no cross-reactivity against other plant RIPs; and has a sensitivity comparable to other reported capture ELISAs using polyclonal antibodies as either capture or detector. MDPI 2017-10-18 /pmc/articles/PMC5666375/ /pubmed/29057799 http://dx.doi.org/10.3390/toxins9100328 Text en © 2017 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Tam, Christina C.
Cheng, Luisa W.
He, Xiaohua
Merrill, Paul
Hodge, David
Stanker, Larry H.
A Monoclonal–Monoclonal Antibody Based Capture ELISA for Abrin
title A Monoclonal–Monoclonal Antibody Based Capture ELISA for Abrin
title_full A Monoclonal–Monoclonal Antibody Based Capture ELISA for Abrin
title_fullStr A Monoclonal–Monoclonal Antibody Based Capture ELISA for Abrin
title_full_unstemmed A Monoclonal–Monoclonal Antibody Based Capture ELISA for Abrin
title_short A Monoclonal–Monoclonal Antibody Based Capture ELISA for Abrin
title_sort monoclonal–monoclonal antibody based capture elisa for abrin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5666375/
https://www.ncbi.nlm.nih.gov/pubmed/29057799
http://dx.doi.org/10.3390/toxins9100328
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