Three monoclonal antibodies (Mabs) to solubilized porcine zona pellucida (s-PZP) were produced. They reacted exclusively to porcine and human zonae pellucidae (ZPe) and two of them blocked sperm penetration of ZP in humans by collaborating with goat anti-mouse gamma-globulin serum as a second antibody. The antigen corresponding to one of these Mabs (Mab 3A4-2G1) was fractionated from s-PZP by immunoaffinity chromatography bound Mab 3A4-2G1. The antigen corresponding to Mab 3A4-2G1 was further fractionated by SDS-PAGE and the protein band corresponding to 92kDa was found to react to Mab 3A4-2G1, as demonstrated by Western blotting. The 92kDa fraction of s-PZP was isolated and injected into mice with complete and incomplete Freund's adjuvants to obtain an antiserum. It was shown that the antiserum to the 92kDa molecule reacted in a tissue-specific manner to human ZP and exhibited a strong inhibitory effect on sperm penetration of ZP of human oocytes.
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http://dx.doi.org/10.1016/0165-0378(88)90037-x | DOI Listing |
J Reprod Immunol
February 1991
Department of Obstetrics and Gynecology, Hyogo Medical College, Japan.
Biochemical and molecular analyses were performed for the porcine zona pellucida (ZP) antigen corresponding to the monoclonal antibody (Mab) 3A4-2G1 exclusively cross-reactive with porcine and human zonae pellucidae (ZPe). By SDS-PAGE and Western blot analysis, Mab 3A4-2G1 reacted with a 92-kDa component of solubilized porcine ZP (s-PZP) under non-reducing conditions, while reacting with a 23-kDa component under reducing conditions. Diagonal SDS-PAGE revealed that the 92-kDa component was composed of 69 kDa and 23 kDa molecules.
View Article and Find Full Text PDFJ Reprod Immunol
October 1988
Department of Obstetrics and Gynecology, Hyogo Medical College, Nishinomiya, Japan.
Three monoclonal antibodies (Mabs) to solubilized porcine zona pellucida (s-PZP) were produced. They reacted exclusively to porcine and human zonae pellucidae (ZPe) and two of them blocked sperm penetration of ZP in humans by collaborating with goat anti-mouse gamma-globulin serum as a second antibody. The antigen corresponding to one of these Mabs (Mab 3A4-2G1) was fractionated from s-PZP by immunoaffinity chromatography bound Mab 3A4-2G1.
View Article and Find Full Text PDFThree monoclonal antibodies (Mabs), 3A4-2G1, 1D5-2B7 and 1F2-1B8 were produced against heat-solubilized porcine zona pellucida (ZP). Each Mab stained intact ZP but no other pig tissues using immunofluorescence staining. All three Mabs stained selectively zonae pellucidae (ZPe) from pigs and humans but not from hamsters, rats or mice, and showed no inhibitory effect on sperm binding to human oocytes.
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