Preparative SDS-PAGE Electroelution for Rapid Purification of Alkyl Hydroperoxide Reductase from Helicobacter pylori

• Main Authors: T Mohammadian, M Doosti, M Paknejad, F Siavoshi, S Massarrat
• Format: Article
• Language: English
• Published: Tehran University of Medical Sciences 2010-03-01
• Series: Iranian Journal of Public Health
• Subjects: Alkyl hydroperoxide reductase; AhpC; Electroelution; Helicobacter pylori
• Online Access: http://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/15195.pdf&manuscript_id=15195

"nBackground: Alkyl hydroperoxide reductase (AhpC) of Helicobacter pylori is considered as a diagnostic antigen. There­fore, this antigen can be used to detect H. pylori infection by stool immunoassays such as ELISA. The aim of this study was to simplify the AhpC protein purification procedures."nMethods: For whole cell protein extraction, the bacterial cells were ruptured by octly-β-D glucopyranoside. The isolation and purification of  AhpC protein were attempted by various techniques including ammonium sulfate precipitation, dialysis, preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electroelution."nResults:A simple method was used for protein purification AhpC protein. One-dimensional preparative gel electrophoresis allows a single and short purification step; the high resolution capacity of this technique leads to a high level of purity of the protein. Moreover, it avoids contamination by other non-specific proteins which often appear during protein purification by column chromatography."nConclusion: The present method is simple, rapid and makes it possible to preparate AhpC from H. pylori