Minimal motif mapping of a known epitope on human zona pellucida protein-4 using a peptide biosynthesis strategy

An important step in the development of a human zona pellucida (huZP) peptide vaccine is to define the minimal amino acid motif for a mapped B cell epitope peptide within huZP4. Identification of this minimal motif is necessary to remove an overlapping T cell epitope that induces a pathogenic T cell response. Here we describe motif (PLTLEL(314-319)) mapping of an 18mer B cell epitope peptide(308-325) on huZP4 protein (previously known as huZP1/ZPB protein), achieved using a set of 22 biosynthetic 8mer peptides fused with truncated glutathione S-transferase (GST) or truncated streptavidin protein, and detected using rabbit anti-porcine zona pellucida (pZP) IgG. The immunogenicity of the B cell epitope peptide was evaluated in rabbits using expressed B cell epitope peptide fused with truncated streptavidin as the antigen. This construct elicited high titer antibody to the 18mer B cell epitope peptide, with reactivity to native human ZP, the biosynthetic 18mer peptide and the 18mer peptide GST fusion protein.