Immunoreactivity evaluation of a new recombinant chimeric protein against Brucella in the murine model
Abdollahi، Abbas نويسنده Department of Microbiology, Fasa University of Medical Sciences, Fars, Iran Abdollahi, Abbas , Mansouri، Shahla نويسنده , , Amani، Jafar نويسنده , , Fasihi-Ramandi، Mahdi نويسنده Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Fasihi-Ramandi, Mahdi , Moradi، Mohammad نويسنده ,
Background and Objectives: Brucellosis is an important health problem in developing countries and no vaccine is available for the prevention of infection in humans. Because of clinically infectious diseases and their economic consequences in human and animals, designing a proper vaccine against Brucella is desirable. In this study, we evaluated the immune responses induced by a designed recombinant chimera protein in murine model.
Materials and Methods: Three immunodominant antigens of Brucella have been characterized as potential immunogenic and protective antigens including: trigger factor (TF), Omp31 and Bp26 were fused together by EAAAK linkers to produce a chimera (structure were designed in silico), which was synthesized, cloned, and expressed in E. coli BL21 (DE3). The purification of recombinant protein was performed using Ni-NTA agarose. SDS-PAGE and anti-His antibody was used for confirmation purified protein (Western blot). BALB/c immunization was performed by purified protein and adjuvant, and sera antibody levels were measured by ELISA. otted.
Results: SDS-PAGE and Western blotting results indicated the similarity of in silico designing and in vitro experiments. ELISA result proved that the immunized sera of mice contain high levels of antibodies (IgG) against recombinant chimeric protein.
Conclusion: The recombinant chimeric protein could be a potential antigen candidate for the development of a subunit vaccine against Brucella.
Keywords: Brucella, Vaccine, Immunity, Recombinant