Production of IgY in chicken egg yollk against helicobacter pylori urease using UreC recombinant protein

ABSTRACT

Helicobacter pylori is a spiral, microaerophilic gram negative bacterium, that multiplies and causes infection in human gastric mucosal layer. H.pylori infection, followed by destruction of gastric epithelial tissue, leads to gastric chronic inflammation, which can cause gastric and peptic ulcers. New approaches have focused on using specific treatments, such as immunotherapy, to eradicate this infection. Urease, as one of the most important virulent and antigenic factors of the bacterium, is a suitable target for this purpose. This study is aimed at production of specific IgY against urease UreC subunit. In this study, initially for preparing recombinant UreC, after purification of the genomic DNA, ureC gene was amplified by polymerase chain reaction [PCR]. The PCR product was ligated to pET28a. The recombinant protein was expressed followed by transformation of recombinant construct into E.coli BL21DE3. SDS-PAGE analysis was carried out and the recombinant protein was purified by Ni-NTA affinity chromatography. The purified recombinant protein was injected to hens. IgY recovered from egg yolk, was purified by PEG precipitation at >70% purity. The purified IgY was analyzed by ELISA and SDS-PAGE. SDS-PAGE analysis revealed a good expression and >70% purification of the recombinant protein. ELISA observation demonstrated high immunogenicity of the recombinant protein. With a view to higher potential of IgY-HpUc in recognition of UreC subunit, the results are in favour of the oral administration of the IgY obtained from hens immunized by H.pylori may provide a novel approach to the management of H.pylori infections