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Generation and Characterization of Specific Monoclonal Antibodies and Nanobodies Directed Against the ATP-Gated Channel P2X4
The P2X4 channel is involved in different physiological and pathological conditions and functions in the nervous system. Despite the existence of several mouse models for which the expression of the gene was manipulated, there is still little information on the expression of the protein at the cellu...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6861843/ https://www.ncbi.nlm.nih.gov/pubmed/31798414 http://dx.doi.org/10.3389/fncel.2019.00498 |
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author | Bergmann, Philine Garcia de Paco, Elvira Rissiek, Björn Menzel, Stephan Dubberke, Gudrun Hua, Jennifer Rassendren, François Ulmann, Lauriane Koch-Nolte, Friedrich |
author_facet | Bergmann, Philine Garcia de Paco, Elvira Rissiek, Björn Menzel, Stephan Dubberke, Gudrun Hua, Jennifer Rassendren, François Ulmann, Lauriane Koch-Nolte, Friedrich |
author_sort | Bergmann, Philine |
collection | PubMed |
description | The P2X4 channel is involved in different physiological and pathological conditions and functions in the nervous system. Despite the existence of several mouse models for which the expression of the gene was manipulated, there is still little information on the expression of the protein at the cellular level. In particular, supposedly specific available antibodies have often proved to recognize unrelated proteins in P2X4-deficient mice. Here, we used an in vivo DNA vaccine approach to generate a series of monoclonal antibodies and nanobodies specific for human, mouse, and rat P2X4 channels. We further characterized these antibodies and show that they solely recognize the native form of the proteins both in biochemical and cytometric applications. Some of these antibodies prove to specifically recognize P2X4 channels by immunostaining in brain or sensory ganglia slices, as well as at the cellular and subcellular levels. Due to their clonality, these different antibodies should represent versatile tools for further characterizing the cellular functions of P2X4 in the nervous system as well as at the periphery. |
format | Online Article Text |
id | pubmed-6861843 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-68618432019-12-03 Generation and Characterization of Specific Monoclonal Antibodies and Nanobodies Directed Against the ATP-Gated Channel P2X4 Bergmann, Philine Garcia de Paco, Elvira Rissiek, Björn Menzel, Stephan Dubberke, Gudrun Hua, Jennifer Rassendren, François Ulmann, Lauriane Koch-Nolte, Friedrich Front Cell Neurosci Cellular Neuroscience The P2X4 channel is involved in different physiological and pathological conditions and functions in the nervous system. Despite the existence of several mouse models for which the expression of the gene was manipulated, there is still little information on the expression of the protein at the cellular level. In particular, supposedly specific available antibodies have often proved to recognize unrelated proteins in P2X4-deficient mice. Here, we used an in vivo DNA vaccine approach to generate a series of monoclonal antibodies and nanobodies specific for human, mouse, and rat P2X4 channels. We further characterized these antibodies and show that they solely recognize the native form of the proteins both in biochemical and cytometric applications. Some of these antibodies prove to specifically recognize P2X4 channels by immunostaining in brain or sensory ganglia slices, as well as at the cellular and subcellular levels. Due to their clonality, these different antibodies should represent versatile tools for further characterizing the cellular functions of P2X4 in the nervous system as well as at the periphery. Frontiers Media S.A. 2019-11-12 /pmc/articles/PMC6861843/ /pubmed/31798414 http://dx.doi.org/10.3389/fncel.2019.00498 Text en Copyright © 2019 Bergmann, Garcia de Paco, Rissiek, Menzel, Dubberke, Hua, Rassendren, Ulmann and Koch-Nolte. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Cellular Neuroscience Bergmann, Philine Garcia de Paco, Elvira Rissiek, Björn Menzel, Stephan Dubberke, Gudrun Hua, Jennifer Rassendren, François Ulmann, Lauriane Koch-Nolte, Friedrich Generation and Characterization of Specific Monoclonal Antibodies and Nanobodies Directed Against the ATP-Gated Channel P2X4 |
title | Generation and Characterization of Specific Monoclonal Antibodies and Nanobodies Directed Against the ATP-Gated Channel P2X4 |
title_full | Generation and Characterization of Specific Monoclonal Antibodies and Nanobodies Directed Against the ATP-Gated Channel P2X4 |
title_fullStr | Generation and Characterization of Specific Monoclonal Antibodies and Nanobodies Directed Against the ATP-Gated Channel P2X4 |
title_full_unstemmed | Generation and Characterization of Specific Monoclonal Antibodies and Nanobodies Directed Against the ATP-Gated Channel P2X4 |
title_short | Generation and Characterization of Specific Monoclonal Antibodies and Nanobodies Directed Against the ATP-Gated Channel P2X4 |
title_sort | generation and characterization of specific monoclonal antibodies and nanobodies directed against the atp-gated channel p2x4 |
topic | Cellular Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6861843/ https://www.ncbi.nlm.nih.gov/pubmed/31798414 http://dx.doi.org/10.3389/fncel.2019.00498 |
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