Usefulness of IgY technology for hepatitis B virus diagnosis

Abstract

an (ELISA) for of affinity chromatography. IgY anti-HBs using polyacrylamide gel electrophoresis (SDS-PAGE), dot blot and enzyme-linked immunosorbent assay (ELISA). Total protein concentrations were measured using spectrophotometer. Samples presenting higher total protein concentration were evaluated as capture protein for in house ELISA to detect HBsAg. Results: The characterization of IgY revealed bands of stained peptides with molecular weight between 75 and 50kDa and 37 and 25kDa using SDS Page. In the dot blot test was observed that there was antigen-antibody reaction throughout the sample period. Mean ± standard deviation (SD) of concentration of total protein obtained after purification by affinity chromatography was 0.386 mg / mL ± 0.440 mg/mL. After evaluation of different dilutions of IgY as capture protein for ELISA, it was possible to distinguish positive from negative controls that presented mean ±SD of optical density (OD) values of 1.99 ± 0.18 and 0.45 ± 0.06, respectively. Conclusion : The results indicated that HBV is able to generate a high production of specific immunoglobulins in chickens and these proteins could be used as capture protein for ELISA. Further studies will be conducting to develop in house ELISA for hepatitis B diagnosis.