Production of immunoglobulin Y (IgY) against synthetic peptide analogs of the immunogenic epitopes of the hepatitis B surface antigen

ABSTRACT

INTRODUCTION: Several studies have been conducted on the use of Immunoglobulin Y (IgY) technology in the fields of diagnostics and therapeutics. IgY is the avian counterpart of the mammalian immunoglobulin G (IgG) which is exclusively transferred from the hen to the yolk thus conferring passive immunization to the growing embryo. However, despite the advantages it offers over the use of mammalian immunoglobulin, IgY technology has remained underutilized. OBJECTIVE:The objective of this study is to produce an IgY with activity against synthetic peptide analogs of known immunogenic epitopes of the Hepatitis B Surface Antigen (HBsAg) - a molecular marker of Hepatitis B infection. METHODS: Chickens were immunized with synthetic peptide analogs of previously reported immunogenic epitopes of the S and the pre-S1 regions of the Hepatitis B surface antigen (HBsAg). IgY specific for the synthetic peptides was isolated by delipidation and salt precipitation and was further purified by affinity chromatography. Purity and molecular weights of the whole IgY molecule and its subunits were assessed and determined by SDS-PAGE. Anti-peptide activity and specificity were determined by indirect ELISA. The study was approved by the Ethical Review Board (ERB) and Technical Review Board of the Research Implementation and Development Office (RIDO), University of the Philippines Manila. RESULTS AND CONCLUSION: The IgY that was purified in this study had an approximate molecular weight of 165 kilodaltons. The heavy and light chains are 60 and 28 kilodaltons, respectively. The affinity purified IgY demonstrated anti-peptide antibody activity against synthetic peptide analogs of known immunogenic epitopes of the HBsAg. Specific binding against a battery of synthetic peptides also revealed that the affinity purified IgY specifically binds to the known immunogenic epitope of the HBsAg.