A Streamlined Method to Obtain Biologically Active TcdA and TcdB Toxins from Clostridioides difficile - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement Access content directly
Journal Articles Toxins Year : 2024

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

Abstract

The major virulence factors of Clostridioides difficile (C. difficile) 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 C. difficile or performed in heterologous hosts. Therefore, we propose a streamlined method to obtain functional toxins in C. difficile. Two C. difficile strains were generated, each harboring a sequence encoding a His-tag at the 3′ end of C. difficile 630∆erm tcdA or tcdB genes. Each toxin gene is expressed using the Ptet 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.

Dates and versions

hal-04417180 , version 1 (25-01-2024)

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Diane Sapa, Anaïs Brosse, Héloïse Coullon, Gauthier Péan de Ponfilly, Thomas Candela, et al.. A Streamlined Method to Obtain Biologically Active TcdA and TcdB Toxins from Clostridioides difficile. Toxins, 2024, 16 (1), pp 38. ⟨10.3390/toxins16010038⟩. ⟨hal-04417180⟩
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