Immunogenicity of a 92kDa component of porcine zona pellucida isolated using a monoclonal antibody (3A4-2G1) exclusively cross-reactive with porcine and human zonae pellucidae.
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.
Hasegawa
A
A
Department of Obstetrics and Gynecology, Hyogo Medical College, Nishinomiya, Japan.
Koyama
K
K
Isojima
S
S
D000818:Animals; D000911:Antibodies, Monoclonal; D003429:Cross Reactions; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D010063:Ovum / Q000276:immunology*; D011247:Pregnancy; D013045:Species Specificity; D013084:Sperm-Ovum Interactions; D013552:Swine; D015044:Zona Pellucida / Q000276:immunology*
D000911:Antibodies, Monoclonal