<i>In silico</i> design of a novel chimeric <i>shigella</i> IpaB fused to C terminal of <i>clostridium perfringens</i> enterotoxin as a vaccine candidate

This study aimed to design a novel chimeric protein <i>in silico</i> to serve as a serotype-independent vaccine candidate against <i>Shigella</i>. The chimera contains amino acid residues 240–460 of <i>Shigella</i> invasion plasmid antigen B (IpaB) and the C-terminus of <i>Clostridium perfringens</i> enterotoxin (C-CPE). Amino acid sequences of 537 peptide linkers were obtained from two protein linker databases. 3D structures of IpaB-CPE_290–319, IpaB-CPE_184–319, IpaB-CPE_194–319 and 537 newly designed IpaB-linker-CPE_290–319 constructs with varying linker regions were predicted. These predicted 3D structures were merged with the 3D structures of native IpaB_240–460, CPE_194–319, CPE_184–319 and CPE_290–319 to select the structure most similar to native IpaB and C-CPE. Several <i>in silico</i> tools were used to determine the suitability of the selected IpaB-C-CPE structure as a vaccine candidate. None of the 537 linkers was capable of preserving the native structure of CPE_290–319 within the IpaB-linker-CPE_290–319 structure. <i>In silico</i> analysis determined that the IpaB-CPE_194–319 3D structure was the most similar to the 3D structure of the respective native CPE domain and that it was a stable chimeric protein exposing multiple B-cell epitopes. IpaB-CPE_194–319 was designed for its capability to bind to human intestinal epithelial and M cells and to accumulate on these cells. The predicted B-cell epitopes are likely to be capable of inducing a mucosal antibody response in the human intestine against <i>Shigella</i> IpaB. This study also showed that the higher binding affinities of CPE_184–319 and CPE_194–319 to claudin molecules than those of CPE_290–319 is the result of preserving the 3D structures of CPE_184–319 and CPE_194–319 when they are linked to the C-termini of other proteins.