Mapping of Epitopes Relevant for Induction of Acrosome Reaction on Human Zona Pellucida Glycoprotein-4 Using Monoclonal Antibodies

作者：Xu Wan Xiang; Bhandari Beena; He Ya Ping; Tang Hai Ping; Chaudhary Sanchita; Talwar Pankaj; Gupta Satish K; Wang Jian^*

来源：American Journal of Reproductive Immunology, 2012, 68(6): 465-475.

DOI：10.1111/j.1600-0897.2012.01177.x

• Acrosome reaction
• epitope mapping
• human zona pellucida glycoprotein-4
• monoclonal antibodies
• recombinant proteins
• peptides

Problem To decipher structural and functional aspects of human zona pellucida glycoprotein-4 (ZP4), the epitopes recognized by monoclonal antibodies (MAbs) have been mapped. Method of study Recombinant human ZP4-mediated induction of acrosome reaction in human sperm was studied in the absence and presence of ZP4-specific MAbs. The epitopes of MAbs were mapped using recombinant peptides expressed in Escherichia coli. Results Monoclonal antibodies (MA-1662, MA-1671) against human ZP4 showed specific binding to ZP matrix of human eggs in an indirect immunofluorescence assay. Both the antibodies showed significant (P < 0.05) inhibition in the baculovirus-expressed recombinant ZP4-mediated acrosome reaction. MA-1671 recognized N-terminal fragment of ZP4 and minimal epitope mapped to amino acid residues 126130 (PARDR), whereas MA-1662 reacted to C-terminal fragment and minimal epitope mapped to amino acid residues 256260 (ENELV). Conclusions The epitopes corresponding to both N- and C-terminal parts of human ZP4 may be relevant for its biological activity.

• 出版日期2012-12
• 单位上海市计划生育科学研究所