A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum

• Main Authors: Lorena Vázquez-Iglesias, Borja Estefanell-Ucha, Leticia Barcia-Castro, María Páez de la Cadena, Paula Álvarez-Chaver, Daniel Ayude-Vázquez, Francisco Javier Rodríguez-Berrocal
• Format: Article
• Language: English
• Published: PeerJ Inc. 2017-06-01
• Series: PeerJ
• Subjects: Antibodies; Slot blot; Alpha-toxin; Electroelution; Clostridium septicum
• Online Access: https://peerj.com/articles/3407.pdf

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.