Background: Tetanus is a kind of neurological disease characterized by rigidity and spasm of muscles. The causative agent of this fatal disease is known as Tetanus Toxin, Clostridium tetani. The Tetanospasmin consists of three chains: H_N, H_C and L. The H_C chain is binds domain and immunogenic part of toxin, so it is proposed as a vaccine candidate. In this study, the recombinant H_C protein obtained from synthetic H_C gene was investigated for its immunogenicity. Additionally, antiserum and polyclonal antibody was prepared against this agent.

Materials and Methods: Recombinant His-tagged Hc protein was expressed in Escherichia coli Bl21 DE3 and purified with Ni-NTA column. Then, rHc protein was used for animal immunization. Along with survey of antibody titer, serum polyclonal antibody was purified by G column.

Results: After gene optimization, expression and purification, rH_C protein was confirmed by western blot technique. Immunization of mice showed high titer of antibody against recombinant protein. Antibody purification from immunized animal serum was carried out as well.

Conclusion: The recombinant H_C protein production and its immunogenicity investigation showed high titer of antibody against Tetanospasmin.