الفهرس | Only 14 pages are availabe for public view |
Abstract Helicobacter pylori bacteria causes gastritis ,duodenal ulcer, and gastric cancer, In present study, we prepare soluble H. pylori antigen, and resolved in 12 % one dimensional SDSPAGE. The resolved polypeptides were stained with coomassie blue stain. Western blot analysis by using specific antiH. pylori antibody identified a sharp band at molecular weight of 58kDa which isolated and purified from H. pylori infected serum samples by electroelution.. The purified 58kDa H. pylori antigen was precipitate with trichloroacetic acid (TCA) and the precipitate appeared as a single band at 58kDa on the coomassie blue stained polyacrylamide gel (SDSPAGE) indicating that the isolated antigen was highly purified. The activity of the purified H. pylori antigen with specific antiH. pylori antibody using enzymelinked immunosorbent assay (ELISA) was destroyed after treatment of the purified antigen with either NaOH or HCl, while the reactivity remained intact after treatment with oxidizing agent (Sodium mperiodate) and reducing agent (mercaptoethanol). The purified antigen was precipitated by 40 % TCA and the reconstituted precipitate showed high reactivity as untreated antigen while the supernatant showed no reactivity with the specific antiH. pylori antibody. The 58 kDa H. pylori antigen was stable when it was stored at 20(R@(BC to 37(R@(BC, while the reactivity was lost when the antigen was heated at 56(R@(BC. supernatant showed no reactivity with the specific antiH. pylori antibody. The 58kDa H. pylori antigen was stable when it was stored at 20(R@(BC to 37(R@(BC, while the reactivity was lost when the antigen was heated at 56(R@(BC. These findings confirmed that the purified antigen was a protein in nature without carbohydrate moieties. This protein was destroyed by ga schemotrypsin after incubation for one hour at 37<U+00BA>C. In conclusion a 58kDa H. pylori antigen was identified, isolated and characterized as protein. The 58kDa H. pylori antigen may be of use in immunodiagnosis of H. pylori infection and possibly in immunization against H. pylori as a step towards a vaccine production. |