title: An immunological method to screen sex-specific proteins of bovine sperm.
abstract: This study was designed to identify sex-specific antibodies (SSAb) in rabbit antisera against bovine sex-sorted sperm, and capture sex-specific proteins of bovine X- or Y- proteins by SSAb. The rabbit antisera against bovine X- or Y-sperm were first produced by a series of immunological approaches, and further purified through immuno-neutralization with excess sex-sorted Y- or X-sperm, respectively, to remove non-sex specific antibodies and enrich sex-specific antibodies. After removal of non-sex specific antibodies, the purified rabbit sera with enriched sex-specific antibodies were screened for sex-specific antibodies by immunofluorescence staining and flow cytometry. The results showed that 3.0, 2.2, and 4.2% of unsorted sperm, sex-sorted X-sperm, and sex-sorted Y-sperm were recognized by the purified rabbit antisera against Y-sperm, respectively, whereas 29.2, 19.7, and 3.9% of unsorted sperm, sex-sorted X-sperm, and sex-sorted Y-sperm were recognized by the purified rabbit antisera against X-sperm. These results suggested that the purified rabbit antisera against X-sperm contained SSAb that preferentially bound to sex-sorted X-sperm. Subsequently, the purified rabbit antisera against X- or Y-sperm were used to immunoprecipitate sex-specific proteins in bovine sperm proteins, and a 30-kDa protein was specifically captured by the rabbit antisera against X-sperm. In conclusion, our results implied that this 30-kDa protein might be a sex-specific protein in bovine X-sperm, which has the potential to be used in immunological procedures for sexing sperm.
authors.affiliation: Key Laboratory for Agricultural Animal Genetics, Breeding, and Reproduction, under Education Ministry of China, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, PR China.
authors.forename: L
authors.initials: L
authors.lastname: Sang
authors.forename: W C
authors.initials: WC
authors.lastname: Yang
authors.forename: L
authors.initials: L
authors.lastname: Han
authors.forename: A X
authors.initials: AX
authors.lastname: Liang
authors.forename: G H
authors.initials: GH
authors.lastname: Hua
authors.forename: J J
authors.initials: JJ
authors.lastname: Xiong
authors.forename: L J
authors.initials: LJ
authors.lastname: Huo
authors.forename: L G
authors.initials: LG
authors.lastname: Yang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis; D002417:Cattle; D007106:Immune Sera; D008297:Male; D008565:Membrane Proteins / Q000276:immunology* / Q000302:isolation & purification; D011817:Rabbits; D012743:Sex Preselection / Q000379:methods / Q000662:veterinary*; D013094:Spermatozoa / Q000276:immunology*; D014960:X Chromosome / Q000276:immunology*; D014998:Y Chromosome / Q000276:immunology*
chemical_list: D000906:Antibodies; D007106:Immune Sera; D008565:Membrane Proteins