Preparation And In Vitro Studies Of Egg Yolk Immunoglobulin(IgY) Against Recombinant VacA-HpaA In Helicobacter Pylori By Gene-Engineering

PART1 PREPARATION OF EGG YOLK IMMUNOGLOBULIN(IGY) AGAINST RECOMBINANT VACA-HPAA IN HELICOBACTER PYLORI BY GENE-ENGINEERINGObjective: To observe high titer of VacA-HpaA IgY from eggs yolk of hens immunized with recombinant VacA-HpaA in Helicobacter pylori by gene-engineering ,which would lay a foundation for the development of treatmental antibody for H.pylori infection.Methods: A mass of recombinant VacA-HpaA protein were aquired by culturing and inducing the engineered bacteria E.coli DH5a contained PQE30-VacA-HpaA vector in the expression of IPTG .The recombinant protein was analyzed by SDS-PAGE and purified by Ni2+-NAT chromatography. Its protein content was detected by Bradford.Its antigenicity and immunogenicity were comfired by ELISA.VacA-HpaA IgY was preparated by immunized hens with the purified recombinant protein (VacA-HpaA) . The feasibility of its clinical application with comparative evaluation was done by H.pylori diagnostic standard of reseach. The relation of IgY titer and immune-time was assessed roughly.