PURIFICATION OF ANTI-HBsAg FROM EGG YOLKS OF IMMUNIZED HENS AND ITS APPLICATION FOR DETECTION OF HBsAg

Background- The hen’s egg yolk possesses an immunoglobulin known as immunoglobulin (Ig) Y, with an advantage over conventional antibody production of certain antigens. The aims of this study were to produce anti-hepatitis B surface antigen (HBsAg) in the egg yolk of hens immunized with hepatitis B vaccine and to coat anti-HBsAg to the wells of an enzyme-linked immunosorbent assay (ELISA) plate for detection of HBsAg in the sera of patients with hepatitis B virus infection. Methods- A group of five hens were immunized intramuscularly with 10 μg of recombinant hepatitis B vaccine at zero, 21 and 80 days. Two weeks after the third dose of the vaccine, the egg yolks were collected. The anti-HBsAg (IgY) was purified using a twostep polyethylene glycol 6000 and one-step 4-M ammonium sulfate process, followed by column purification. The purified anti-HBsAg (5 μg/mL) was coated to the wells of ELISA plates for detection of HBsAg. Results- Two weeks after the third dose of vaccine, all the hens’ sera were positive for anti-HBsAg (IgY). The sensitivity and specificity were 73.1% and 85.7%, respectively. Conclusion- HBV vaccine was immunogenic in hens and ELISA using the purified anti- HBsAg from the egg yolks was able to detect the HBsAg in the sera of patients infected with hepatitis B virus.