Purpose of Mycobacterium tuberculosis ERP protein as a recombinant and prevalence of an immunity response analysis

BCG is the only vaccine against tuberculosis caused by Mycobacterium tuberculosis bacteria. Effective new tuberculosis vaccines need to be developed because this vaccine is insufficient in adults. In this thesis, erp gene from Mycobacterium tuberculosis strain isolated from a tuberculosis patient in Turkey was cloned into pGEM-T, followed by pET28-a vector and transferred to E. coli BL21 strain. His-tagged recombinant Erp protein expressed in E. coli was purified using nickel columns and the vaccine formulations were prepared by mixing 7: 3 with Montanide ISA 720 VG, an oil-based adjuvant. The antigenic properties of the recombinant protein were qualitatively visualized by Western blotting. When using recombinant protein and BCG vaccine-specific serum, the expected large bands were observed, but no band was observed when only adjuvant-specific serum was used. Simultaneously, the humoral and cellular immunity triggering capacity of the recombinant protein was assessed. When comparing pre- and post-immunization IgG values, it was determined that the prepared vaccine formulation significantly increased the humoral immune response. The cellular immunity triggering capacities of this formulation were measured by serum interferon-gamma (IFN-γ) and interleukin-12 (IL-12) levels. Erp formulation was found to be as effective as BCG in increasing IFN-γ levels. Erp formulation was found to provide higher levels of IL-12 than BCG on days 15 and 30. It is thought that recombinant ERP can be used to develop the subtype tuberculosis vaccine in the direction of the obtained findings.