Usage of Peptide Antigens for Antibody-Based BoNT Detection System

Abstract

Objective: In the current study, we aimed to elicit a BoNT-specific immune response via using peptides designed by using bioinformatics tools instead of using intact native BoNT A toxin itself and to demonstrate the usage of these anti-peptide antibodies in a toxin detection system. Methods: Synthetic epitopic regions specific to LC, HN and HC regions of BoNT A were selected as antigen using B cell “Epitope Prediction Tools” form IEDB and shown to be on the surface of BoNT A with a software (Discovery Studio 4.0). Selected peptides were used in mice immunizations and the interaction between developed anti-peptide antibodies and the native intact BoNT A was examined. Results: The detection of native intact BoNT A at picogram levels per milliliter was performed with anti-peptide antibodies developed against three different peptides (P1, P2, and P3) in mice. Conclusion: The current study shows that synthetic peptides are at least as effective as the native toxin or the toxoid itself for raising high-affinity antibodies against toxins. In addition, considering the need for a quick diagnosis of botulism and, already used test systems in which many experimental animals are sacrificed, these results demonstrate the necessity of synthetic peptide immunogens usage to reduce both the number of animals and the amount of toxin usage.