Materials and Methods: ID-LFn was generated by overlapping PCR followed by cloning in pET28a. The recombinant protein was then expressed and purified by Ni-NTA chromatography. Reactivity of ID-LFn with anti-PA/LF/EF antibodies was checked by ELISA. Stability was assessed using Circular Dichroism Spectroscopy. The vaccine potential of ID-LFn was evaluated by toxin neutralization assay, lymphocyte proliferation assay, and cytokine analysis. The protection efficacy was analyzed by challenge studies in mice.
Results: ID-LFn was found to be significantly stable as compared to protective antigen. Anti-ID-LFn antibodies recognized PA, LF as well as EF. Though, the total antibody titre, toxin neutralization activity was found to be less than PA but surprisingly, the protection efficacy of ID-LFn was found similar as PA.
Conclusion: The ID-LFn vaccine might be second next generation vaccine showing equal protection but higher shelf life as PA with the capability of neutralizing PA, LF as well as EF at the same time. Thus, it may prove an efficient and reliable treatment strategy against anthrax.
R. Bhatnagar, None