Evaluation of multi-epitope recombinant protein as a candidate for a contraceptive vaccine

2021 ◽

Vol 19 (1) ◽

Author(s):

Nathaniel Philip Sandam ◽

Dhamodhar Prakash ◽

Prashanth Thimmareddy

Keyword(s):

Recombinant Protein ◽

Gnrh Receptor ◽

Colony Pcr ◽

E Coli ◽

Contraceptive Vaccine ◽

Sds Page ◽

High Antibody Titer ◽

Stray Dog ◽

Contraceptive Vaccines ◽

Antigenic Epitopes

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.

Download Full-text

Contraceptive vaccine bounces back

Nature India ◽

10.1038/nindia.2010.102 ◽

2010 ◽

Keyword(s):

Contraceptive Vaccine

Download Full-text

Effectiveness of zona pellucida protein ZPB as an immunocontraceptive antigen

Reproduction ◽

10.1530/reprod/120.1.19 ◽

2000 ◽

pp. 19-32 ◽

Cited By ~ 19

Author(s):

ML Martinez ◽

JD Harris

Keyword(s):

Menstrual Cycle ◽

Recombinant Protein ◽

Antibody Titre ◽

Zona Pellucida ◽

Protein A ◽

Papio Cynocephalus ◽

Cynomolgus Monkeys ◽

Menstrual Cycles ◽

Normal Menstrual Cycle

Immunization of female mammals with native zona pellucida (ZP) proteins is known to cause infertility. Since each human ZP protein is now available as a purified recombinant protein, is it possible to compare the immunocontraceptive potential of each ZP protein. A breeding study was conducted in cynomolgus monkeys (Macaca fasicularis) after immunization with recombinant human ZP (rhZP) proteins (ZPA, ZPB, ZPC) separately and in combinations. This study demonstrated that immunization with recombinant human ZPB (rhZPB) protein caused cynomolgus monkeys to become infertile for 9-35 months. A second study was conducted in baboons (Papio cynocephalus), which yielded a similar result. The baboons immunized with rhZPB became infertile for 9 to > 20 months. During the time of maximum antibody titre, some animals experienced disruption of the menstrual cycle, but eventually all of the animals resumed normal menstrual cycles. Control animals and animals immunized with other rhZP proteins all became pregnant before any of the rhZPB-treated animals. This is the first study in which a recombinant ZP protein has consistently induced infertility in a primate without permanent disruption of the normal menstrual cycle.

Download Full-text

Differences between antigenic determinants of pig and cat zona pellucida proteins

Reproduction ◽

10.1530/reprod/119.1.15 ◽

2000 ◽

pp. 15-23 ◽

Cited By ~ 15

Author(s):

K Jewgenow ◽

M Rohleder ◽

I Wegner

Keyword(s):

In Vitro Fertilization ◽

Zona Pellucida ◽

Antigenic Determinants ◽

Vitro Fertilization ◽

Contraceptive Vaccine ◽

Sperm Binding ◽

Dependent Inhibition ◽

Dose Dependent Inhibition ◽

Dose Dependent

Despite many efforts, the control of reproduction in feral cat populations is still a problem in urban regions around the world. Immunocontraception is a promising approach; thus the present study examined the suitability of the widely used pig zona pellucida proteins (pZP) for contraception in feral domestic cats. Purified zona pellucida proteins obtained from pig and cat ovaries were used to produce highly specific antisera in rabbits. Antibodies against pZP raised in rabbits or lions were not effective inhibitors of either in vitro sperm binding (cat spermatozoa to cat oocytes) or in vitro fertilization in cats, whereas antibodies against feline zona pellucida proteins (fZP) raised in rabbits showed a dose-dependent inhibition of in vitro fertilization. Immunoelectrophoresis, ELISA and immunohistology of ovaries confirmed these results, showing crossreactivity of anti-fZP sera to fZP and to a lesser extent to pZP, but no interaction of anti-pZP sera with fZP. It is concluded that cat and pig zonae pellucidae express a very small number of shared antigenic determinants, making the use of pZP vaccine in cats questionable. A contraceptive vaccine based on feline zona pellucida determinants will be a better choice for the control of reproduction in feral cats if immunogenity can be achieved.

Download Full-text

Use of Lectins for Analysis of Glycans in Recombinant Protein Biopharmaceuticals

Российский иммунологический журнал ◽

10.31857/s102872210002646-5 ◽

2018 ◽

pp. 707-709

Author(s):

V. Monakhova ◽

M. Sushkin ◽

N. Pigareva ◽

A. Simbirtsev

Keyword(s):

Recombinant Protein

Download Full-text

Immunoenhancement of Recombinant Neisseria meningitides PorB Protein on Porcine Circovirus Type 2 and Mycoplasma hyopneumoniae Genetically Engineered Vaccines

Protein and Peptide Letters ◽

10.2174/0929866526666190430115052 ◽

2019 ◽

Vol 26 (10) ◽

pp. 776-784

Author(s):

Rui Yang ◽

Yu Tao ◽

Gaojian Li ◽

Jian Chen ◽

Jianhong Shu ◽

...

Keyword(s):

Recombinant Protein ◽

Mycoplasma Hyopneumoniae ◽

Porcine Circovirus Type ◽

Porcine Circovirus Type 2 ◽

Genetically Engineered ◽

Porcine Circovirus ◽

Practical Applications ◽

Neisseria Meningitides ◽

Short Period

Background:Porcine circovirus and Mycoplasma hyopneumoniae can cause respiratory diseases in pigs, which cause serious economic loss in the worldwide pig industry. Currently, these infections are mainly prevented and controlled by vaccination. The new vaccines on the market are mainly composed of subunits and inactivated vaccines but usually have lower antigenicity than traditional live vaccines. Thus, there is an increasing need to develop new adjuvants that can cause rapid and long-lasting immunity to enhance the antigenic efficacy for vaccines. Studies have shown that meningococcal porin PorB can act as a ligand to combine with Toll-like receptors to activate the production of immunological projections and act as a vaccine immunological adjuvant.Objective:In this article, we expressed and purified the recombinant PorB protein and verified its immunogenicity against porcine circovirus type 2 and Mycoplasma hyopneumoniae genetically engineered vaccine.Methods:In this article, we used prokaryotic expression to express and purify recombinant PorB protein, four different concentrations of PorB protein, Freund's adjuvant with two genetically engineered vaccines were combined with subcutaneous immunization of mice.Results:Our study shows that the appropriate dose of the recombinant protein PorB can enhance the levels of humoral and cellular responses induced by two genetically engineered vaccines in a short period of time in mice. The PorB adjuvant group may cause statistically higher antibody titers for both genetically engineered vaccines compared to Freund's commercial adjuvant (P<0.001).Conclusion:The recombinant protein PorB may be a good candidate adjuvant for improving the protective effect of vaccines against porcine circovirus type 2 and Mycoplasma hyopneumoniae, and the protein can be used for future practical applications.

Download Full-text