Production of specific IgY Helicobacter pylori recombinant OipA protein and assessment of its inhibitory effects towards attachment of H. pylori to AGS cell line

• Published in: Clinical and experimental vaccine research (Seoul) 2015-07, Vol.4 (2), p.177-183
• Main Authors: Borhani, Katayoun, Mobarez, Ashraf Mohabati, Khabiri, Ali Reza, Behmanesh, Mehrdad, Khoramabadi, Nima
• Format: Article
• Language: English
• Subjects: Antibiotics; Antibodies; Gastric cancer; Immunization; Immunoglobulins; Laboratory animals; Molecular weight; Original; Polyethylene glycol; Proteins; Virulence
• ISSN: 2287-3651; 2287-366X
• DOI: 10.7774/cevr.2015.4.2.177

The common triple therapy for Helicobacter pylori is challenged by the increasing cases of antibiotic resistant infections, raising the need to explore alternative therapies. Oral administration of egg yolk immunoglobulin Y (IgY) has been previously reported as a means of passive immunization therapy for H. pylori infections. In this work, we investigated the inhibitory effect of IgY on the attachment of H. pylori to AGS cell line. Recombinant OipA was prepared. Hens were immunized with recombinant protein three times. IgY was purified from egg yolks of immunized hens using polyethylene glycol precipitation method. The inhibitory effect of the specific immunoglobulin was evaluated in AGS cell line infected with H. pylori. The presence of recombinant OipA (30 kD) was confirmed via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunization of hens was confirmed using enzyme-linked immunosorbent assay. The purified IgY from egg yolks were assessed using SDS-PAGE and confirmed by western blot. The results showed that IgY-OipA had inhibitory effect on attachment of H. pylori to AGS cell line and may be utilized as a therapeutic or prophylaxis material.