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Journal of the Society for Gynecologic Investigation
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Generation and Characterization of Antibodies Against Synthetic Peptides of Porcine Zona Pellucida ZP3{alpha}

Lori-Linell H. Hall, MD

Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, Michigan; Department of Obstetrics, Gynecology, and Reproductive Sciences, Magee-Womens Hospital, 300 Halket Street, Pittsburgh, PA 15213

Edward C. Yurewicz, PhD

Anthony G. Sacco, PhD

Kamran S. Moghissi, MD

Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, Michigan

Objective: Use of synthetic zona peptides as target immunogens is a promising approach to an anti-fertility vaccine. To elucidate contraceptive epitopes, we raised polyclonal antibodies against synthetic peptides of pig ZP3{alpha} and evaluated for immunogenicity, ability to inhibit in vitro boar sperm-pig zona attachment, and cross-reactivity with human zonae.

Methods: Five synthetic peptides were chosen based on hydropathicity analysis of ZP3{alpha}, synthesized and coupled to keyhold limpet haemocyanin (KLH). Pairs of male rabbits were immunized with each peotide-KLH conjugate using a multi-site intradermal injection protocol. Resulting antisera were evaluated for immunoreactivity with cognate peptide and ZP3{alpha}/EBGD (purified endo-beta-galactosidase-digested ZP3{alpha}), contraceptive potential using an in vitro pig gamete bioassay, and cross-reactivity with human zonae using indirect immunofluorescence.

Results: All anti-peptide-KLH sera demonstrated immunoreactivity with cognate peptide. Anti-peptide sera, except for anti-{alpha}-3-KLH serum, were less immunoreactive with ZP3{alpha}/EBGD. Antisera directed against {alpha}-1-KLH, {alpha}-2-KLH, and {alpha}-3-KLH demonstrated significant (P < .00001) inhibition of boar sperm-pig zona attachment. Only anti-{alpha}-2-KLH and anti-{alpha}-3-KLH sera cross-reacted with human zonae.

Conclusion: Synthetic peptides of pig ZP3{alpha} are immunogenic, elicit antibodies cross-reactive with human zonae, and have in vitro contraceptive activity. These data support continued investigation of synthetic ZP3{alpha} peptides as potential target immunogens for anti-fertility vaccine development.

Key Words: Zona pellucida • synthetic zona peptides • pig ZP3{alpha} • anti-fertility vaccine

Journal of the Society for Gynecologic Investigation, Vol. 2, No. 3, 552-558 (1995)
DOI: 10.1177/107155769500200308
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