POLYCLONAL ANTIBODY PRODUCTION AND PURIFICATION AGAINST TET-ANOSPASMIN

Shahram Nazarian Golamreza Olad Mohhamad Ali Arefpour Jafar Salimian Mohhamd Javad Bagheripour Mohhamd Ebrahim Minaie Ehsan Rezaie Assistant Professor, Biology Research Centre, Imam Hossein University, Tehran, Iran. PhD, Molecular Biotechnology, Applied Biotechnology Research Centre, Baqitatolah University, Tehran, Iran. MSc, Cell and Molecular Biology, Biology Research Centre, Imam Hossein University, Tehran, Iran. PhD, Immunology, Chemical Injuries Research Center, Baqitatolah University, Tehran, Iran. PhD Student, Nanobiotechnology, Biology Research Centre, Imam Hossein University, Tehran, Iran. MSc, Cell and Molecular Biology, Applied Biotechnology Research Centre, Baqitatolah University, Tehran, Iran. Corresponding Author: Mohhamad Ali Arefpour Tel: (+98)2171104934 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: HN, HC and L. The HC chain is binds domain and immunogenic part of toxin, so it is proposed as a vaccine candidate. In this study, the recombinant HC protein obtained from synthetic HC 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, rHC 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 HC protein production and its immunogenicity investigation showed high titer of antibody against Tetanospasmin.