Purification an‎d Characterization of Epsilon an‎d Beta Toxins of Clostridium perfringens Type B using Liquid Chromatography to Prepare Specific antibodies against each Toxin

10/22/2026 12:00:00 AM

Clostridium perfringens strain B, which simultaneously produces potent epsilon (ETX) an‎d beta (CPB) toxins, is the main cause of the fatal disease enterotoxemia in livestock. Rapid diagnosis an‎d effective treatment of this disease require access to purified toxins an‎d specific antibodies. In this thesis, a single-step purification protocol was designed an‎d optimized. In this process, a single anion exchange chromatography (AEX) step was used to separate both toxins. The purity an‎d identity of the toxins were confirmed using size exclusion chromatography, SDS-PAGE, an‎d mass spectrometry, an‎d their biological activity was eva‎luated in an animal model. The single-step anion exchange chromatography strategy successfully isolated an‎d purified epsilon toxin with a purity of 99% an‎d beta toxin with a purity of over 94%. Immunization with the prepared toxoids resulted in the production of high-titer antisera with appropriate specificity against both toxins. Finally, this thesis successfully presented a comprehensive protocol for the production of pure an‎d active toxins, which provides a valuable infrastructure for the development of accurate diagnostic kits, vaccine quality control, an‎d fundamental studies.

كلستريديوم پرفرنجنس , توكسين اپسيلون , توكسين بتا , انتروتوكسمي , كروماتوگرافي تبادل آنيوني , كروماتوگرافي اندازه طردي , خالص‌سازي پروتئين , آنتي‌بادي پلي‌كلونال

Clostridium perfringens , Epsilon toxin , Beta toxin , Enterotoxemia , Anion-Exchange Chromatography , Size-Exclusion Chromatography , Protein Purification , Polyclonal Antibody

Seyed Amir Hossein Fallah Javadi Nezhad Lialestani

Afsaneh Mollahosseini, Ali Es-haghi