1887

Abstract

To explore an epitope-based vaccine against , we screened the epitopes in the N2N3 subdomain of fibronectin-binding protein A (FnBPA) as a surface component of .

We expressed N2N3 proteins and prepared monoclonal antibodies (mAbs) against N2N3 by the hybridoma technique, before screening the B-cell epitopes in N2N3 using a phage-displayed random 12-mer peptide library with these mAbs against N2N3. Finally, we analysed the characters of the screened epitopes using immunofluorescence and an infection assay.

In this paper, we identified a linear B-cell epitope in N2N3 through screening a phage-displayed peptide library with a 3C3 mAb against the N2N3. The 3C3 mAb recognized the IETFNKANNRFSH sequence of the N2N3 subdomain. Subsequently, site-directed mutagenic analysis demonstrated that residues F162, K164, N167, R168 and F169 formed the core of IETFNKANNRFSH, and this core motif was the minimal determinant of the B-cell epitope recognized by the 3C3 mAb. The epitope IETFNKANNRFSH showed high homology among different strains. Moreover, this epitope was exposed on the surface of the by using an enzyme-linked immunosorbent assay (ELISA) assay and an indirect immunofluorescence assay. As expected, the epitope peptide evoked a protective immune response against infection in immunized mice.

We identified a novel linear B-cell epitope, IETFNKANNRFSH, in the N2N3 subdomain of fibronectin-binding protein A that is recognized by 3C3 mAb, which will contribute to the further study of an epitope-based vaccine candidate against .

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2018-03-01
2020-04-07
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