[Recombinant expression of Clostridium perfringens toxins and preparation of monoclonal antibodies].

This study aimed to construct prokaryotic expression systems for six major toxins of Clostridium perfringens and obtain specific monoclonal antibodies, thereby providing reliable experimental tools for toxin detection and early diagnosis of related diseases. Six kings of major toxin genes of C. perfringens were cloned, and related recombinant fusion protein expression vectors were constructed. Recombinant proteins were expressed through prokaryotic systems and purified. The purified recombinant proteins served as immunogens for animal immunization to generate monoclonal antibody-producing hybridoma cell lines. The specificity of the antibodies against both recombinant and native toxins was verified by Western blotting. Six recombinant fusion proteins were successfully expressed and purified, and three monoclonal antibody-producing hybridoma cell lines were established after subcloning, designated as 3B10 (targeting CPA), 10E9 (targeting ETX), and 5F9 (targeting NetB) respectively. Western blotting results demonstrated that these antibodies specifically recognized both recombinant and native toxins, exhibiting strong immunoreactivity. In conclusion, specific monoclonal antibodies against three major C. perfringens toxins were successfully generated, and it may provide valuable biological resources and effective tools for rapid toxin detection and early diagnosis of C. perfringens infection in goats in future, with significant potential for practical application.