Background: The role of autoantibodies against zona pellucida (ZP) in infertility and success of IVF is debatable. In the present study, a novel recombinant human zona protein is used to address this issue.
Materials and methods: Fourteen serum samples positive for porcine ZP, six from infertile patients, and 20 from healthy women were analysed. Sera were tested for their reactivity against a recombinant multi-epitope peptide (named ZPCP7) composed of ten predictable antigenic peptides from human ZP proteins by ELISA. In addition, seven identified fine epitope peptides (fused with a GST188 carrier protein) of human ZP3 and ZP4 were used as antigens for western blotting.
Results and discussion: Eight of 14 porcine ZP antibody positive sera and one out of six testing infertile patients showed positive reaction in ELISA when all sera were tested at 1:2560 dilution. Further, out of nine serum samples seven also showed reactivity to three to five of epitope peptides fused with truncated GST188 carrier protein in western blot respectively. Thus, seven sera samples were finally determined as ZP antibody positive in our two steps method of analysis. Also, the positive serum sample (no. S3) was able to recognize native human ZP in indirect immunofluorescence experiment. These results provide evidence that ZP antibodies are associated with infertility. The use of recombinant multi-epitope peptide based on human zona proteins may also overcome the problem of false positivity observed using porcine ZP as an antigen.
02 - 04 Sep 2014
World Congress of Reproductive Biology