一种用于快速蛋白质纯化的简单电洗脱方法：从……中分离α毒素并制备抗体
A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from .
作者信息
Vázquez-Iglesias Lorena, Estefanell-Ucha Borja, Barcia-Castro Leticia, Páez de la Cadena María, Álvarez-Chaver Paula, Ayude-Vázquez Daniel, Rodríguez-Berrocal Francisco Javier
机构信息
Department of Biochemistry, Genetics and Immunology, Facultad de Biología, Universidad de Vigo, Vigo, Spain.
Unidad de Proteómica, Servicio de Determinación Estructural, Proteómica y Genómica, CACTI, Universidad de Vigo, Spain.
出版信息
PeerJ. 2017 Jun 22;5:e3407. doi: 10.7717/peerj.3407. eCollection 2017.
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 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 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.
它会在人类和家畜中引发多种疾病，在大多数情况下，如果没有临床干预，这些疾病都是致命的。α毒素是一种重要的致病因子，是由（此处原文缺失相关信息）产生的独特致死性毒力因子。这种毒素具有溶血、高致死性和坏死活性，但正被用作抗原以开发动物疫苗。本研究的目的是分离（此处原文缺失相关信息）的α毒素并产生针对它的高度特异性抗体。在这项工作中，我们基于电洗脱开发了一种简单有效的α毒素纯化方法，该方法可作为一种节省时间的蛋白质纯化方法。该技术避免了其他蛋白质纯化技术（如色谱法）可能出现的其他蛋白质污染。高度纯化的毒素用于产生多克隆抗体。通过蛋白质免疫印迹法检测抗体的特异性，这些抗体可用于通过斑点印迹法定量测定α毒素。
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