Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Fine Epitope Mapping of the Vibrio cholera Toxins A, B, and P and an ELISA Assay

Version 1 : Received: 18 July 2022 / Approved: 21 July 2022 / Online: 21 July 2022 (10:32:40 CEST)

How to cite: De-Simone, S.G.; Napoleão-Pêgo, P.; Gonçalves, P.S.; Lechuga, G.C.; Cardozo, S.V.; Provance, D.W.; Morel, C.M.; Silva, F.R. Fine Epitope Mapping of the Vibrio cholera Toxins A, B, and P and an ELISA Assay . Preprints 2022, 2022070320 (doi: 10.20944/preprints202207.0320.v1). De-Simone, S.G.; Napoleão-Pêgo, P.; Gonçalves, P.S.; Lechuga, G.C.; Cardozo, S.V.; Provance, D.W.; Morel, C.M.; Silva, F.R. Fine Epitope Mapping of the Vibrio cholera Toxins A, B, and P and an ELISA Assay . Preprints 2022, 2022070320 (doi: 10.20944/preprints202207.0320.v1).

Abstract

Oral immunization with the choleric toxin (CT) elicits a high level of protection against its enterotoxin activities and can control cholera in endemic settings. However, the complete B-cell epitope map of the CT responsible for protection remains to be clarified. Here, we have mapped the B-cell linear epitopes of the three chains of the CT protein (CTP) prepared by Spot synthesis. The immunoreactivity of sera from mice immunized with an oral, inactivated vaccine (Schankol†™) was measured against membrane-bound peptides for mapping. Results: Eighteen IgG epitopes were identified; eight in the CTA, three in the CTB, and seven in the protein P. Three epitopes, TQTGFVRHDDGYVST (aa 66-77, Vc/TxA-3), KNGAIFQVE VPGSQN (aa 64-78, Vc/TxB-11), and LNDEHK (aa 90-95, Vc/TxP-16), were chosen to synthesize a multiple antigen peptide that was used to coat ELISA plates to screen immunized mouse sera as a test for an in vitro diagnostics for cholera. Conclusion: Vaccination with inactive CT-generated antibodies against multiple linear epitopes and the selected epitopes can be used to construct immunological reagents for rapid serological diagnosis of cholera with high sensitivity and specificity.

Keywords

Cholera toxin; B-cell epitope; peptide microarray; synthetic peptide; MAPs; ELISA

Subject

LIFE SCIENCES, Microbiology

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