Molecular Biology Reports

, Volume 41, Issue 1, pp 217–223

Tentative identification of sex-specific antibodies and their application for screening bovine sperm proteins for sex-specificity

  • Wu-Cai Yang
  • Lei Sang
  • Yao Xiao
  • Hua-Lin Zhang
  • Ke-Qiong Tang
  • Li-Guo Yang
Article

Abstract

Our previous studies indicated that a purified rabbit antiserum against X-sperm contained sex-specific antibodies (SSAbs) which preferentially bound to sex-sorted X-sperm. The specificity of sex-specific antiserum was initially demonstrated using flow cytometry only, which resulted in uncertainty. In this study, the putative SSAbs against bovine X-sperm (XSSAb) were produced by a series of immunological approaches, and the effectiveness of separation of sperm using putative XSSAb was validated. Subsequently, the XSSAb was used to immunoprecipitate sex-specific proteins (SSPs) in bovine sperm, followed by two-dimensional gel electrophoresis. The results showed 7.6, 15.2 and 52.1 % of sex-sorted Y-sperm, sex-sorted X-sperm and unsorted sperm were recognized by the neutralized rabbit antisera against X-sperm, respectively. Also the purity of separation of sperm using putative XSSAb reached 74.3 % when the immunologically separated sperm were injected into oocytes. In addition, three candidate SSP sports about 30 kDa were captured by the XSSAb. Our results confirmed that the putative XSSAb contained SSAbs, and implied that these three protein sports might be SSPs in bovine X-sperm. This provides a potentially more efficient method for sorting sperm and lays a foundation for future search for SSPs.

Keywords

Sex-specific antibodies Sex-specific proteins 2-DE Sexing 

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Wu-Cai Yang
    • 1
    • 2
  • Lei Sang
    • 3
  • Yao Xiao
    • 2
  • Hua-Lin Zhang
    • 2
  • Ke-Qiong Tang
    • 2
  • Li-Guo Yang
    • 2
  1. 1.College of Animal Science and TechnologyNorthwest A&F UniversityShaanxiChina
  2. 2.College of Animal Science and TechnologyHuazhong Agricultural UniversityWuhanChina
  3. 3.Institute of Animal Husbandry and Veterinary MedicineFujian Academy of Agricultural SciencesFuzhouChina

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