Recombinant expression of in silico identified B-cell epitope of epsilon toxin of Clostridium perfringens in translational fusion with a carrier protein

Epsilon toxin secreted by Clostridium perfringens types B and D has been directly implicated as the causative agent of fatal enterotoxemia in domestic animals. The aim of the present study is to use in silico approach for identification of B-cell epitope (s) of epsilon toxin, and its expression in fusion with a carrier protein to analyze its potential as vaccine candidate (s). Using different computational analyses and bioinformatics tools, a number of antigenic determinant regions of epsilon toxin were identified. One of the B cell epitopes of epsilon toxin comprising the region (amino acids 40-62) was identified as a promising antigenic determinant. This Etx epitope (Etx sub(40-62)) was cloned and expressed as a translational fusion with B-subunit of heat labile enterotoxin (LTB) of E. coli in a secretory expression system. Similar to the native LTB, the recombinant fusion protein retained the ability to pentamerize and bind to GM sub(1) ganglioside receptor of LTB. The rLTB. Etx sub(40-62) could be detected both with anti-Etx and anti-LTB antisera. The rLTB. Etx sub(40-62)). fusion protein thus can be evaluated as a potential vaccine candidate against C. perfringens.