A Streamlined Method to Obtain Biologically Active TcdA and TcdB Toxins from .

The major virulence factors of () are enterotoxins A (TcdA) and B (TcdB). The study of toxins is a crucial step in exploring the virulence of this pathogen. Currently, the toxin purification process is either laborious and time-consuming in or performed in heterologous hosts. Therefore, we propose a streamlined method to obtain functional toxins in . Two strains were generated, each harboring a sequence encoding a His-tag at the 3' end of 630∆ or genes. Each toxin gene is expressed using the P promoter, which is inducible by anhydro-tetracycline. The obtained purification yields were 0.28 mg and 0.1 mg per liter for rTcdA and rTcdB, respectively. In this study, we successfully developed a simple routine method that allows the production and purification of biologically active rTcdA and rTcdB toxins with similar activities compared to native toxins.