Investigation of the inhibitory effects of immunoglobulin Y antibody against key epitopes of Helicobacter pylori UreB recombinant protein

Abstract

Helicobacter pylori (H.pylori) is considered to be the most important gastrointestinal pathogen causing gastritis, gastric ulcers and even gastric cancer. The treatment of these infections has failed due to the rapidly increasing antibiotic resistance to standard treatment regimens and the lack of an effective vaccine. This study investigates the production and therapeutic potential of Immunoglobulin Y (IgY) antibodies targeting key epitopes of the H. pylori UreB recombinant protein. Given the increasing challenge of antibiotic resistance in H. pylori treatment, this research underscores the necessity for alternative therapeutic strategies. A specific region of the UreB gene, containing critical immunogenic epitopes, was amplified using Polymerase Chain Reaction (PCR) and cloned into the pET32b vector. The recombinant plasmid was expressed in Escherichia coli BL21 (DE3), and the UreB protein was purified via Ni-NTA affinity chromatography, confirmed by SDS-PAGE and Western blot analysis. Hens were immunized with the recombinant UreB protein, resulting in the generation of specific IgY antibodies. The purified IgY-UreB antibodies exhibited a remarkable reduction in urease activity by 84.53 % at a concentration of 10 mg/mL, effectively neutralizing this critical virulence factor. Additionally, in vitro assays demonstrated that IgY-UreB antibodies significantly inhibited the growth of H. pylori at a concentration of 5 mg/mL. These findings highlight the potential of IgY as a viable alternative to traditional antibiotic therapies, particularly in the context of rising antibiotic resistance. This study paves the way for the development of innovative immunotherapeutic strategies that may improve treatment outcomes for H. pylori infections.

Keywords: Gastritis; Gene cloning; Helicobacter pylori; IgY antibodies; Multi epitope; Urease B.