Production of anti-Helicobacter pylori urease-specific immunoglobulin in egg yolk using an antigenic epitope of H. pylori urease.
The potential therapeutic effects of Helicobacter pylori-specific immunoglobulin (IgY-Hp) derived from egg yolk and identification of the immunodominant H. pylori proteins have previously been reported. In this study, the urease epitope that is recognized by IgY-Hp was identified and used as an immunogen to produce urease-specific IgY (IgY-HpU). Epitope regions were mapped and peptides of selected epitope regions were synthesized. The IgY-Hp titre against synthetic peptides was evaluated using ELISA analysis. Hens were immunized with synthetic peptides conjugated with BSA. Urease activity was quantified by measuring the optical density of an indicator dye. Of the five synthetic peptides assayed, a peptide representing 15 amino acid residues of UreB (UB-3; aa 396-410, DNDNFRIKRYLSKYT) was specifically recognized by the IgY-Hp. Immunization of hens with BSA-conjugated UB-3 resulted in the generation of IgY-HpU. IgY-HpU markedly reduced H. pylori urease activity by 80 % as compared to control IgY (IgY-BSA). The availability of the synthetic UreB-derived peptide enabled the production of highly specific anti-urease IgY, which had a significant inhibitory effect on H. pylori urease activity. Therefore, specific IgY-HpU produced using the synthetic peptide may be an effective tool against infection by H. pylori.
Shin
Ji-Hyun
JH
Department of Pharmacology1 and Gastroenterology2, Dankook University College of Medicine, Cheonan, Korea.
Roe
Im-Hwan
IH
Department of Pharmacology1 and Gastroenterology2, Dankook University College of Medicine, Cheonan, Korea.
Kim
Hyung-Gun
HG
Department of Pharmacology1 and Gastroenterology2, Dankook University College of Medicine, Cheonan, Korea.
D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology; D002645:Chickens; D004530:Egg Yolk / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D005260:Female; D016480:Helicobacter pylori / Q000201:enzymology / Q000276:immunology*; D016056:Immunodominant Epitopes / Q000276:immunology*; D007136:Immunoglobulins / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D010446:Peptide Fragments / Q000276:immunology; D014510:Urease / Q000276:immunology*
D000942:Antigens, Bacterial; C016512:IgY; D016056:Immunodominant Epitopes; D007136:Immunoglobulins; D010446:Peptide Fragments; D014510:Urease