Contraceptive efficacy of recombinant fusion protein comprising zona pellucida glycoprotein-3 fragment and gonadotropin releasing hormone

Ananta Prasad Arukha, Vidisha Minhas, Abhinav Shrestha, Satish Kumar Gupta

Research output: Contribution to journalArticlepeer-review

13 Scopus citations


Contraceptive vaccines have been used for the management of wildlife population. In the present study, we have examined the contraceptive potential of Escherichia coli-expressed recombinant fusion protein comprising of 'promiscuous' T cell epitope of tetanus toxoid [TT; amino acid (aa) residues 830-844] followed by dilysine linker (KK), dog ZP3 fragment (aa residues 307-346), triglycine spacer (GGG), T cell epitope of bovine RNase (bRNase; aa residues 94-104), GnRH, T cell epitope of circumsporozoite protein of Plasmodium falciparum (CSP; aa residues 362-383), and GnRH. SDS-PAGE analysis of the purified refolded protein revealed a dominant ~12 kDa band, which in Western blot reacted with mouse polyclonal antibodies against dog ZP3 fragment and mouse monoclonal antibodies against GnRH. Immunization of female FvB/J mice following two booster schedule with the above recombinant protein supplemented with alum led to high antibody titres against the immunogen as well as ZP3 and GnRH as determined by ELISA. The immune sera reacted with zona pellucida of mouse oocyte and also inhibited in-vitro fertilization. The qRT-PCR studies showed decrease in the ovarian GnRH receptor in mice immunized with the recombinant fusion protein. Mating studies revealed high contraceptive efficacy of the recombinant protein as in two independent experiments, 90% of the immunized female mice failed to conceive. Following one booster immunization schedule, 50% of the immunized female mice failed to conceive. However, in adjuvanted controls, all the female mice became pregnant. To conclude, the recombinant protein described herein has a good potential to be developed as candidate contraceptive vaccine.

Original languageEnglish (US)
Pages (from-to)18-26
Number of pages9
JournalJournal of Reproductive Immunology
StatePublished - Apr 1 2016
Externally publishedYes

Bibliographical note

Funding Information:
This work was funded by the financial support from Department of Biotechnology (DBT), Government of India (BT/PR5207/MED/15/81/2012) and National Institute of Immunology, New Delhi . SKG would like to acknowledge Tata Innovation Fellowship awarded to him by DBT. The funding agencies had no role in study design, collection, analysis or interpretation of the data.

Publisher Copyright:
© 2016 Elsevier Ireland Ltd.


  • Gonadotropin releasing hormone
  • Immunocontraception
  • Promiscuous T cell epitope
  • Tag-free recombinant fusion protein
  • Zona pellucida glycoprotein-3


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