تخليص ايمونوگلوبين )TGY(Y با سه روش كروماتوگرافي تعويض يون ، كروماتوگرافي جذبي ، و رسوب دهي با پلي اتيلن گليكول

Purification of Egg Yolk Immunoglobulin (IgY) by Three Methods; Affinity Chromatography, Ion Exchange Chromatography and Poly Ethylene Glycol Precipitation.

Introduction: Egg yolk is a rich and available source of immunoglobulin Y (IgY) that can be used in medical diagnosis and treatment against microbial agents. Methods and Materials: In this study anti-human IgG specific IgY was produced in hens and purified from egg yolk using three different methods. After immunization of hens against human IgG, IgY was extracted from egg yolk by the acidic water dilution method and purified using three methods; polyethylene glycol precipitation, ion exchange chromatography and affinity chromatography. Molecular mass and activity of the purified antibody were estimated using SDS-PAGE and ELISA, respectively. Results: The results showed that the affinity produced IgY antibodies has purity and yield of 95% and 78% respectively. In addition, the suitable and simple method for purification of IgY fraction was prepared by PEG precipitation with recovery and purity of 90% and 95% respectively. Ion exchange chromatography product had recovery and purity of 72% and 68%. Furthermore the molecular weight (MW) of intact IgY and its light and heavy chains were estimated to be 190, 27 and 67 KD, respectively. This product can be used for diagnosis of IgG in different diagnostic tests.