Preparation and purification of anti immunoglobulin Y antibodies from rabbit

Background and Objective: Chicken egg yolk antibodies (IgY) play an important role as an alternative to mammalian polyclonal antibodies. They are recently used in medical research for prophylaxis, diagnosis and treatment of diseases. Anti-IgY antibodies are used for immunoassay and therapy. The aim of this study was preparation and purification of anti-immunoglobulin Y antibodies from rabbit serum and confirm of its activity. Methods: IgY and other water-soluble proteins were extracted with acid and chloroform and then thiophilic chromatography (T-gel) was performed and applied for purification of IgY. Two female rabbits were immunized with IgY in Freud’s adjuvant and then booster doses were administered, blood samples were collected by times. Ammonium sulfate precipitation, then gel filtration or T-gel chromatography used for purification of Anti-IgY antibodies. Cellulous acetate electrophoresis, double immunodiffusion and ELISA procedures used for confirm of identification and activity. Findings: The purified protein (IgY) was observed in the γ-globulin region of electrophoresis. The purified antibodies from rabbit serum were observed in the γ-globulin region, which reacted against IgY in double immunodiffusion and the titer of purified antibody-using ELISA, 1.10000 obtained. Conclusion: These antibodies serve as the basis of many immunoassays for detection and assay of several molecules as anti-immunoglobulin Y reagents.