A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum
Clostridium septicum produces a number of diseases in human and farm animals which, in most of the cases, are fatal without clinical intervention. Alpha toxin is an important agent and the unique lethal virulent factor produced by Clostridium septicum. This toxin is haemolytic, highly lethal and nec...
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doaj-5e3d54fc1077465ab010c10c50e8ecc32020-11-25T00:00:40ZengPeerJ Inc.PeerJ2167-83592017-06-015e340710.7717/peerj.3407A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicumLorena Vázquez-Iglesias0Borja Estefanell-Ucha1Leticia Barcia-Castro2María Páez de la Cadena3Paula Álvarez-Chaver4Daniel Ayude-Vázquez5Francisco Javier Rodríguez-Berrocal6Department of Biochemistry, Genetics and Immunology, Facultad de Biología, Universidad de Vigo, Vigo, SpainDepartment of Biochemistry, Genetics and Immunology, Facultad de Biología, Universidad de Vigo, Vigo, SpainDepartment of Biochemistry, Genetics and Immunology, Facultad de Biología, Universidad de Vigo, Vigo, SpainDepartment of Biochemistry, Genetics and Immunology, Facultad de Biología, Universidad de Vigo, Vigo, SpainUnidad de Proteómica, Servicio de Determinación Estructural, Proteómica y Genómica, CACTI, Universidad de Vigo, SpainDepartment of Biochemistry, Genetics and Immunology, Facultad de Biología, Universidad de Vigo, Vigo, SpainDepartment of Biochemistry, Genetics and Immunology, Facultad de Biología, Universidad de Vigo, Vigo, SpainClostridium septicum produces a number of diseases in human and farm animals which, in most of the cases, are fatal without clinical intervention. Alpha toxin is an important agent and the unique lethal virulent factor produced by Clostridium septicum. This toxin is haemolytic, highly lethal and necrotizing activities but is being used as an antigen to develop animal vaccines. The aim of this study was to isolate the alpha toxin of Clostridium septicum and produce highly specific antibodies against it. In this work, we have developed a simple and efficient method for alpha toxin purification, based on electroelution that can be used as a time-saving method for purifying proteins. This technique avoids contamination by other proteins that could appear during other protein purification techniques such chromatography. The highly purified toxin was used to produce polyclonal antibodies. The specificity of the antibodies was tested by western blot and these antibodies can be applied to the quantitative determination of alpha toxin by slot blot.https://peerj.com/articles/3407.pdfAntibodiesSlot blotAlpha-toxinElectroelutionClostridium septicum |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Lorena Vázquez-Iglesias Borja Estefanell-Ucha Leticia Barcia-Castro María Páez de la Cadena Paula Álvarez-Chaver Daniel Ayude-Vázquez Francisco Javier Rodríguez-Berrocal |
spellingShingle |
Lorena Vázquez-Iglesias Borja Estefanell-Ucha Leticia Barcia-Castro María Páez de la Cadena Paula Álvarez-Chaver Daniel Ayude-Vázquez Francisco Javier Rodríguez-Berrocal A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum PeerJ Antibodies Slot blot Alpha-toxin Electroelution Clostridium septicum |
author_facet |
Lorena Vázquez-Iglesias Borja Estefanell-Ucha Leticia Barcia-Castro María Páez de la Cadena Paula Álvarez-Chaver Daniel Ayude-Vázquez Francisco Javier Rodríguez-Berrocal |
author_sort |
Lorena Vázquez-Iglesias |
title |
A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum |
title_short |
A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum |
title_full |
A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum |
title_fullStr |
A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum |
title_full_unstemmed |
A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum |
title_sort |
simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from clostridium septicum |
publisher |
PeerJ Inc. |
series |
PeerJ |
issn |
2167-8359 |
publishDate |
2017-06-01 |
description |
Clostridium septicum produces a number of diseases in human and farm animals which, in most of the cases, are fatal without clinical intervention. Alpha toxin is an important agent and the unique lethal virulent factor produced by Clostridium septicum. This toxin is haemolytic, highly lethal and necrotizing activities but is being used as an antigen to develop animal vaccines. The aim of this study was to isolate the alpha toxin of Clostridium septicum and produce highly specific antibodies against it. In this work, we have developed a simple and efficient method for alpha toxin purification, based on electroelution that can be used as a time-saving method for purifying proteins. This technique avoids contamination by other proteins that could appear during other protein purification techniques such chromatography. The highly purified toxin was used to produce polyclonal antibodies. The specificity of the antibodies was tested by western blot and these antibodies can be applied to the quantitative determination of alpha toxin by slot blot. |
topic |
Antibodies Slot blot Alpha-toxin Electroelution Clostridium septicum |
url |
https://peerj.com/articles/3407.pdf |
work_keys_str_mv |
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