Production and Characterization of Recombinant Multi-Epitope G Glycoprotein of Rabies Virus

Maha Gasmi; Ziad Rahma; Ruth Williams-Hooker Williams-Hooker; Aastha Sharma

doi:10.55940/medphar202351

Authors

Maha Gasmi Manouba University
Ziad Rahma Microbiology Department, University of Constantine 1, Algeria.
Ruth Williams-Hooker College of Health Sciences, University of Memphis, Zach Curlin Drive, Memphis, TN 38152, USA
Aastha Sharma Department of engineering and science, University- GD Goenka University Gurugram, India.

DOI:

https://doi.org/10.55940/medphar202351

Keywords:

Epitopes, G Glycoprotein, Immunization, Rabies, Vaccine

Abstract

Background: Therapeutic proteins have gained increasing interest in research and development for treatments and vaccines against numerous diseases in recent years. This study focuses on producing a recombinant multi-epitope protein of the G glycoprotein, which will serve as a novel vaccine candidate against rabies virus.

Aim: This research aimed to consolidate various 'neutralizing' B epitopes into a single protein and investigate its biochemical and biological characteristics.

Material and Method: The recombinant multi-epitope G glycoprotein was synthesized after transforming the E. coli BL21 bacterial strain with the genetic construct. The protein was produced in soluble form following bacterial induction with 1 mM IPTG (Isopropyl β-D-1-thiogalactopyranoside) and purified using a Ni-NTA affinity column. The 'multi-epitope' protein was characterized by 12% SDS-PAGE. The specificity of binding of the novel recombinant G glycoprotein to human (serum) rabies antibodies was compared to the binding of these same antibodies to whole and inactivated rabies viruses available in the laboratory, namely, the rabies PV-11 strain and the CVS-11 strain, using an indirect ELISA test.

Result: The results demonstrate that the E. coli BL21 strain successfully produced the new recombinant protein with a molecular weight of 30 kDa and that the loss of glycosylation has no effect on the immunogenicity of this protein. Structurally, the new recombinant protein has preserved the epitopes recognized by serum from hyperimmune individuals.

Conclusion: These findings suggest the potential efficacy of the multi-epitope recombinant G glycoprotein as a vaccine candidate and the possibility of developing a novel vaccine based on this multivalent recombinant protein.

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