Preparation,purification and identification of rabbit polyclonal antibody against recombinant protein AP65 of Trichomonas vaginalis

Objective To prepare, purify and identify the rabbit polyclonal antibody against fusion AP65 of Trichomonas vaginalis. Methods The fusion AP65 was used to immunize rabbits for the antibody. The titer of antiserum was demonstrated by indirect ELISA. The polyclonal antibodies were purified by saturation ammonium sulfate and n protein A sepharose 4 FF. The immunogenicity of the ployclonal antibody was determined by western blot. Results The titers of antibody detected by indirection ELISA reached to 1∶25 600. The purification of the polyclonal antibody was highly 80% by n protein A sepharose 4 FF after purified by ammonium sulfate, which was 56% however. The specification was confirmed by western blot. Conclusion The high-purity and high-titer polyclonal antibody was prepared successfully.