Immunocontraceptive potential of a GnRH receptor-based fusion recombinant protein
Abstract Background The management of stray dog population has been of utmost importance due to their overpopulation, increase in dog bites incidence, and rabies. Contraceptive vaccines, a non-surgical alternative to spaying and neutering are viewed as a valuable option for the management of dog pop...
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| Format: | Article |
| Language: | English |
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Elsevier
2021-05-01
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| Series: | Journal of Genetic Engineering and Biotechnology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s43141-021-00164-9 |
| _version_ | 1797204306434195456 |
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| author | Nathaniel Philip Sandam Dhamodhar Prakash Prashanth Thimmareddy |
| author_facet | Nathaniel Philip Sandam Dhamodhar Prakash Prashanth Thimmareddy |
| author_sort | Nathaniel Philip Sandam |
| collection | DOAJ |
| description | Abstract Background The management of stray dog population has been of utmost importance due to their overpopulation, increase in dog bites incidence, and rabies. Contraceptive vaccines, a non-surgical alternative to spaying and neutering are viewed as a valuable option for the management of dog population. In this study, the contraceptive potential of a recombinant fusion protein containing the three genes GnRH, GnRH receptor, and ZP3 was explored. Results The gene fragment encoding GnRH, GnRHR, and ZP3 along with the antigenic epitopes of canine distemper virus and tetanus toxoid was assembled, synthesized, and cloned into pET28a expression vector. The resulting construct GVAC08 was successfully transformed into BL21DE3 strain of E. coli and confirmed by colony PCR. The recombinant GVAC08 protein was expressed and purified using Ni-NTA and was confirmed to be a 50-KDa protein by SDS PAGE and Western blot. Mice were immunized with the GVAC08 protein using Freund’s complete adjuvant followed by a booster using Freund’s incomplete adjuvant. This induced a high antibody titer against GnRH, GnRH receptor, and ZP3 which was determined by ELISA. Conclusion Mating studies showed that the GVAC08 recombinant protein was able to reduce the litter size in immunized mice showing improved efficacy. However, the vaccine candidate with further improvements will be a viable contraceptive vaccine. |
| first_indexed | 2024-04-24T08:33:08Z |
| format | Article |
| id | doaj.art-274b15d14eb94502b27ac61080714048 |
| institution | Directory Open Access Journal |
| issn | 2090-5920 |
| language | English |
| last_indexed | 2024-04-24T08:33:08Z |
| publishDate | 2021-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Genetic Engineering and Biotechnology |
| spelling | doaj.art-274b15d14eb94502b27ac610807140482024-04-16T18:10:09ZengElsevierJournal of Genetic Engineering and Biotechnology2090-59202021-05-011911810.1186/s43141-021-00164-9Immunocontraceptive potential of a GnRH receptor-based fusion recombinant proteinNathaniel Philip Sandam0Dhamodhar Prakash1Prashanth Thimmareddy2Department of Biotechnology, M.S. Ramaiah Institute of TechnologyDepartment of Biotechnology, M.S. Ramaiah Institute of TechnologyGeniron Biolabs Pvt. Ltd.Abstract Background The management of stray dog population has been of utmost importance due to their overpopulation, increase in dog bites incidence, and rabies. Contraceptive vaccines, a non-surgical alternative to spaying and neutering are viewed as a valuable option for the management of dog population. In this study, the contraceptive potential of a recombinant fusion protein containing the three genes GnRH, GnRH receptor, and ZP3 was explored. Results The gene fragment encoding GnRH, GnRHR, and ZP3 along with the antigenic epitopes of canine distemper virus and tetanus toxoid was assembled, synthesized, and cloned into pET28a expression vector. The resulting construct GVAC08 was successfully transformed into BL21DE3 strain of E. coli and confirmed by colony PCR. The recombinant GVAC08 protein was expressed and purified using Ni-NTA and was confirmed to be a 50-KDa protein by SDS PAGE and Western blot. Mice were immunized with the GVAC08 protein using Freund’s complete adjuvant followed by a booster using Freund’s incomplete adjuvant. This induced a high antibody titer against GnRH, GnRH receptor, and ZP3 which was determined by ELISA. Conclusion Mating studies showed that the GVAC08 recombinant protein was able to reduce the litter size in immunized mice showing improved efficacy. However, the vaccine candidate with further improvements will be a viable contraceptive vaccine.https://doi.org/10.1186/s43141-021-00164-9DogsImmunocontraceptionGnRHGnRHRZP3Recombinant fusion protein |
| spellingShingle | Nathaniel Philip Sandam Dhamodhar Prakash Prashanth Thimmareddy Immunocontraceptive potential of a GnRH receptor-based fusion recombinant protein Journal of Genetic Engineering and Biotechnology Dogs Immunocontraception GnRH GnRHR ZP3 Recombinant fusion protein |
| title | Immunocontraceptive potential of a GnRH receptor-based fusion recombinant protein |
| title_full | Immunocontraceptive potential of a GnRH receptor-based fusion recombinant protein |
| title_fullStr | Immunocontraceptive potential of a GnRH receptor-based fusion recombinant protein |
| title_full_unstemmed | Immunocontraceptive potential of a GnRH receptor-based fusion recombinant protein |
| title_short | Immunocontraceptive potential of a GnRH receptor-based fusion recombinant protein |
| title_sort | immunocontraceptive potential of a gnrh receptor based fusion recombinant protein |
| topic | Dogs Immunocontraception GnRH GnRHR ZP3 Recombinant fusion protein |
| url | https://doi.org/10.1186/s43141-021-00164-9 |
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