[Preparation of egg yolk immunoglobulin (IgY) against p53 protein expressed in E.coli].

OBJECTIVE

To isolate immunoglobulin (IgY) from egg yolk of p53 protein-immunized hens and to study its reactivity to the antigens.

METHOD

Immunization of egg-laying hens was performed for 3 times at the interval of 14 days with purified p53 protein that had been expressed in E.coli. The eggs laid by these immunized hens were then collected during the whole period of the experiment to prepare IgY from egg yolk by ammonium sulfate precipitation. Final purification and identification of the IgY were performed using SDS-PAGE, enzyme-linked immunosorbent assay and Western blotting.

RESULT

All immunized hens developed specific antibodies to p53 protein in contrast to the control ones. The highest titers of the IgY occurred 4 weeks after the first immunization and reached 1:10(6), which remained stable for up to 3 months.

CONCLUSION

IgY technology is less costly, non-invasive, fast, simple and highly efficient to generate polyclonal antibodies.