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

PeerJ

article

Lorena Vázquez-Iglesias¹  Borja Estefanell-Ucha¹  Leticia Barcia-Castro¹  María Páez de la Cadena¹  Paula Álvarez-Chaver²  Daniel Ayude-Vázquez¹  Francisco Javier Rodríguez-Berrocal¹ 

[1] Department of Biochemistry, Genetics and Immunology, Facultad de Biología, Universidad de Vigo;Unidad de Proteómica, Servicio de Determinación Estructural, Proteómica y Genómica, CACTI, Universidad de Vigo

关键词: Antibodies;    Slot blot;    Alpha-toxin;    Electroelution;    Clostridium septicum;   

DOI  :  10.7717/peerj.3407

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

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.

【 授权许可】

CC BY