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<Articles><Article><Journal><PublisherName></PublisherName><JournalTitle>DARU Journal of Pharmaceutical Sciences</JournalTitle><Volume>16</Volume><Issue>3</Issue></Journal><ArticleTitle>A simple and cost-effective method for rapid purification of alkyl hydroperoxide reductase (AhpC) from Helicobacter pylori and its antibody production</ArticleTitle><FirstPage>174</FirstPage><LastPage>181</LastPage><AuthorList/><History><PubDate PubStatus="received"><Year>2015</Year><Month>10</Month><Day>06</Day></PubDate></History><Abstract>Background and the purpose of the study: Helicobacter pylori express abundant amounts of AhpC enzyme that functions to reduce organic hydroperoxides (ROOH) into the corresponding non-toxic alcohols (ROH). This conserved antigen has been earlier described as specific and unique for H. pylori and therefore, both H. pylori AhpC and Anti-AhpC could be useful in the development of serologic and stool antigen tests, to detecting and monitoring H. pylori infection. AhpC may also serves as a potential target for an antimicrobial agent or for vaccine development. The aim of this study was to simplify isolation and purification of the AhpC and production of a highly specific polyclonal antibody against it.
Methods and Results: In this paper a simple method was used for protein purification and antibody production which avoids both the long term AhpC protein purification procedure and the addition of Freund's adjuvant. One-dimensional preparative gel electrophoresis allows a single and short purification step and the high resolution capacity of this technique leads to a high level of purity of the protein and consequently to a very high specificity of the antibody. Moreover, it avoids contamination by other non-specific proteins which often appear during protein purification by column chromatographic techniques.
Major Conclusion: The present method is simple, rapid and cost-effective and makes it possible to produce antibody for stool antigen enzyme immunoassay in short time and at low cost.</Abstract><web_url>https://daru.tums.ac.ir/index.php/daru/article/view/364</web_url><pdf_url>https://daru.tums.ac.ir/index.php/daru/article/download/364/364</pdf_url></Article></Articles>
