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Extraction and Inhibition of Enzymatic Activity of Botulinum Neurotoxins/A1, /A2, and /A3 by a Panel of Monoclonal Anti-BoNT/A Antibodies

Botulinum neurotoxins (BoNTs) are extremely potent toxins that are capable of causing death or respiratory failure leading to long-term intensive care. Treatment includes serotype-specific antitoxins, which must be administered early in the course of the intoxication. Rapidly determining human expos...

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Autores principales: Kalb, Suzanne R., Lou, Jianlong, Garcia-Rodriguez, Consuelo, Geren, Isin N., Smith, Theresa J., Moura, Hercules, Marks, James D., Smith, Leonard A., Pirkle, James L., Barr, John R.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2670495/
https://www.ncbi.nlm.nih.gov/pubmed/19399171
http://dx.doi.org/10.1371/journal.pone.0005355
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author Kalb, Suzanne R.
Lou, Jianlong
Garcia-Rodriguez, Consuelo
Geren, Isin N.
Smith, Theresa J.
Moura, Hercules
Marks, James D.
Smith, Leonard A.
Pirkle, James L.
Barr, John R.
author_facet Kalb, Suzanne R.
Lou, Jianlong
Garcia-Rodriguez, Consuelo
Geren, Isin N.
Smith, Theresa J.
Moura, Hercules
Marks, James D.
Smith, Leonard A.
Pirkle, James L.
Barr, John R.
author_sort Kalb, Suzanne R.
collection PubMed
description Botulinum neurotoxins (BoNTs) are extremely potent toxins that are capable of causing death or respiratory failure leading to long-term intensive care. Treatment includes serotype-specific antitoxins, which must be administered early in the course of the intoxication. Rapidly determining human exposure to BoNT is an important public health goal. In previous work, our laboratory focused on developing Endopep-MS, a mass spectrometry-based endopeptidase method for detecting and differentiating BoNT/A–G serotypes in buffer and BoNT/A, /B, /E, and /F in clinical samples. We have previously reported the effectiveness of antibody-capture to purify and concentrate BoNTs from complex matrices, such as clinical samples. Because some antibodies inhibit or neutralize the activity of BoNT, the choice of antibody with which to extract the toxin is critical. In this work, we evaluated a panel of 16 anti-BoNT/A monoclonal antibodies (mAbs) for their ability to inhibit the in vitro activity of BoNT/A1, /A2, and /A3 complex as well as the recombinant LC of A1. We also evaluated the same antibody panel for the ability to extract BoNT/A1, /A2, and /A3. Among the mAbs, there were significant differences in extraction efficiency, ability to extract BoNT/A subtypes, and inhibitory effect on BoNT catalytic activity. The mAbs binding the C-terminal portion of the BoNT/A heavy chain had optimal properties for use in the Endopep-MS assay.
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spelling pubmed-26704952009-04-28 Extraction and Inhibition of Enzymatic Activity of Botulinum Neurotoxins/A1, /A2, and /A3 by a Panel of Monoclonal Anti-BoNT/A Antibodies Kalb, Suzanne R. Lou, Jianlong Garcia-Rodriguez, Consuelo Geren, Isin N. Smith, Theresa J. Moura, Hercules Marks, James D. Smith, Leonard A. Pirkle, James L. Barr, John R. PLoS One Research Article Botulinum neurotoxins (BoNTs) are extremely potent toxins that are capable of causing death or respiratory failure leading to long-term intensive care. Treatment includes serotype-specific antitoxins, which must be administered early in the course of the intoxication. Rapidly determining human exposure to BoNT is an important public health goal. In previous work, our laboratory focused on developing Endopep-MS, a mass spectrometry-based endopeptidase method for detecting and differentiating BoNT/A–G serotypes in buffer and BoNT/A, /B, /E, and /F in clinical samples. We have previously reported the effectiveness of antibody-capture to purify and concentrate BoNTs from complex matrices, such as clinical samples. Because some antibodies inhibit or neutralize the activity of BoNT, the choice of antibody with which to extract the toxin is critical. In this work, we evaluated a panel of 16 anti-BoNT/A monoclonal antibodies (mAbs) for their ability to inhibit the in vitro activity of BoNT/A1, /A2, and /A3 complex as well as the recombinant LC of A1. We also evaluated the same antibody panel for the ability to extract BoNT/A1, /A2, and /A3. Among the mAbs, there were significant differences in extraction efficiency, ability to extract BoNT/A subtypes, and inhibitory effect on BoNT catalytic activity. The mAbs binding the C-terminal portion of the BoNT/A heavy chain had optimal properties for use in the Endopep-MS assay. Public Library of Science 2009-04-28 /pmc/articles/PMC2670495/ /pubmed/19399171 http://dx.doi.org/10.1371/journal.pone.0005355 Text en This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. https://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.
spellingShingle Research Article
Kalb, Suzanne R.
Lou, Jianlong
Garcia-Rodriguez, Consuelo
Geren, Isin N.
Smith, Theresa J.
Moura, Hercules
Marks, James D.
Smith, Leonard A.
Pirkle, James L.
Barr, John R.
Extraction and Inhibition of Enzymatic Activity of Botulinum Neurotoxins/A1, /A2, and /A3 by a Panel of Monoclonal Anti-BoNT/A Antibodies
title Extraction and Inhibition of Enzymatic Activity of Botulinum Neurotoxins/A1, /A2, and /A3 by a Panel of Monoclonal Anti-BoNT/A Antibodies
title_full Extraction and Inhibition of Enzymatic Activity of Botulinum Neurotoxins/A1, /A2, and /A3 by a Panel of Monoclonal Anti-BoNT/A Antibodies
title_fullStr Extraction and Inhibition of Enzymatic Activity of Botulinum Neurotoxins/A1, /A2, and /A3 by a Panel of Monoclonal Anti-BoNT/A Antibodies
title_full_unstemmed Extraction and Inhibition of Enzymatic Activity of Botulinum Neurotoxins/A1, /A2, and /A3 by a Panel of Monoclonal Anti-BoNT/A Antibodies
title_short Extraction and Inhibition of Enzymatic Activity of Botulinum Neurotoxins/A1, /A2, and /A3 by a Panel of Monoclonal Anti-BoNT/A Antibodies
title_sort extraction and inhibition of enzymatic activity of botulinum neurotoxins/a1, /a2, and /a3 by a panel of monoclonal anti-bont/a antibodies
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2670495/
https://www.ncbi.nlm.nih.gov/pubmed/19399171
http://dx.doi.org/10.1371/journal.pone.0005355
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