Molecular cloning of an acrosomal sperm antigen gene and the production of its recombinant protein for immunocontraceptive vaccine.
A monoclonal antibody, HS-63, which reacts specifically with a highly conserved sperm acrosome antigen, was shown to inhibit in vitro fertilization of mouse and human. The corresponding sperm antigen designated as MSA-63 was purified to homogeneity from mouse testes and used as an immunogen to generate polyclonal antisera in rabbits. The cDNA fragments of MSA-63 gene were cloned from mouse testis cDNA library by an immunoscreening method using polyclonal antisera specific for MSA-63. Using the established cDNA clone as a probe, the gene encoding for MSA-63 protein was found to be conserved among different mammalian species. Only one specific mRNA 1.5 kb in size was identified from the adult mouse testis among different mouse tissues. The recombinant fusion protein containing MSA-63 protein fragment was produced in Escherichia coli and used to immunize female mice. Similar to the original HS-63 monoclonal antibody, the antisera thus produced reacted only with the sperm acrosome and revealed significant inhibition to the in vitro fertilization of mouse oocytes. The results of this preliminary study suggest that it is feasible to mass produce sperm-specific antigens or their antigenic fragments by recombinant DNA technology for the development of sperm antigen-based immunocontraceptive vaccines.
Liu
M S
MS
Department of Obstetrics and Gynaecology, University of British Columbia, Vancouver, Canada.
Chan
K M
KM
Lau
Y F
YF
Lee
C Y
CY
D000177:Acrosome / Q000276:immunology*; D000818:Animals; D000941:Antigens / Q000235:genetics*; D003001:Cloning, Molecular; D003267:Contraception; D003269:Contraception, Immunologic; D004247:DNA / Q000235:genetics; D006801:Humans; D008297:Male; D051379:Mice; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015183:Restriction Mapping; D013094:Spermatozoa / Q000276:immunology*; D014612:Vaccines / Q000302:isolation & purification
D000941:Antigens; D011993:Recombinant Fusion Proteins; D014612:Vaccines; D004247:DNA