Localization of epitopes for monoclonal antibodies at the N-terminus of the porcine zona pellucida glycoprotein pZPC

Gupta, S. K. ; Yurewicz, Edward C. ; Afzalpurkar, Abhijit ; Rao, K. V. S. ; Gage, Douglas A. ; Wu, H. ; Sacco, A. G. (1995) Localization of epitopes for monoclonal antibodies at the N-terminus of the porcine zona pellucida glycoprotein pZPC Molecular Reproduction and Development, 42 (2). pp. 220-225. ISSN 1040-452X

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Official URL: http://onlinelibrary.wiley.com/doi/10.1002/mrd.108...

Related URL: http://dx.doi.org/10.1002/mrd.1080420211


Zona pellucida glycoproteins play an important role in fertilization. In this study, attempts have been made to identify and define epitopes of monoclonal antibodies (mAbs) possessing contraceptive efficacy in vitro. The porcine zona glycoprotein pZPC, a homologue of mouse/human ZP3, was reduced and alkylated and subsequently digested with trypsin. Reverse-phase HPLC of the tryptic digest yielded twenty two peaks (T1-T22). When tested against mAbs reactive against sequential determinants on pZPC, T11 was immunoreactive with two mAbs, mAb-455 and mAb-467, as shown by antigen inhibition ELISA. IC50 values of 3.1 nM and 8.6 nM were recorded versus mAb-455 and mAb-467 respectively, and approximated the IC50 values obtained with intact pZPC. Amino acid analysis, Edman degradation, and FAB-MS identified T11 as the N-blocked decapeptide pyro-Gln-Pro-Val-Trp-Gln-Asp-Glu-Gly-Gln-Arg derived from the N-terminus of pZPC. Synthesis of overlapping octapeptides further identified VWQDE and WQDE as the minimum motifs with antigenie activity for mAb-455 and mAb-467, respectively. Glycine replacement peptides confirmed residues W,Q,E as critical for binding mAb-455 and W,Q,D,E as critical for binding mAb-467. Both mAbs inhibited binding of boar sperm to zona-encased porcine oocytes. These results, the first to define peptide epitopes of porcine zona glycoprotein, will assist in the design of an immunocontraceptive vaccine based on synthetic peptides corresponding to pZPC or its homologues in other species.

Item Type:Article
Source:Copyright of this article belongs to John Wiley and Sons.
Keywords:Porcine Zona Pellucida; Sperm-egg Binding; Peptide Immunoassay; Monoclonal Antibody; Epitope Mapping
ID Code:72155
Deposited On:28 Nov 2011 06:09
Last Modified:28 Nov 2011 06:09

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