Methods of producing purified clostridial toxin comprising tangential flow filtration, hydrophobic interaction chromatography and anion exchange chromatography are disclosed. These methods provide good yields of C. difficile toxin having a purity of about 90% or greater. Highly purified Clostridial toxins, toxoids (e.g., prepared by inactivating the toxin as disclosed herein) and compositions comprising these toxins and/or toxoids are also disclosed. Methods of using the purified toxins and/or toxoids for example, to elicit an immune response against clostridium (e.g., C. difficile) are also disclosed.本發明揭示產生經純化之梭菌毒素的方法,該等方法包含切向流過濾、疏水性相互作用層析及陰離子交換層析。此等方法提供純度為約90%或90%以上之困難梭菌毒素的良好產率。亦揭示經高度純化之梭菌毒素、類毒素(例如藉由使如本文所揭示之毒素失活來製備)及包含此等毒素及/或類毒素之組合物。亦揭示使用經純化之毒素及/或類毒素例如誘發針對梭菌屬(例如困難梭菌)之免疫反應的方法。
