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


  • 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.



Epitopes, G Glycoprotein, Immunization, Rabies, Vaccine


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.


Kumar A, Bhatt S, Kumar A, & Rana, T. Canine Rabies: An epidemiological significance, pathogenesis, diagnosis, prevention and public health issues. Comparative Immunology, Microbiology and Infectious Diseases. 2023;101992. DOI:10.1016/j.cimid.2023.101992

Fisher CR, Streicker DG, & Schnell MJ. The spread and evolution of rabies virus: conquering new frontiers. Nature Reviews Microbiology. 2018; 16(4), 241-255. DOI: 10.1038/nrmicro.2018.11

Gasmi M & Rahma Z. THE HISTORY OF RABIES IN TUNISIA AND THE DEVELOPMENT OF THE RABIES VACCINE. Journal of Biological Pharmaceutical And Chemical Research. 2023; 10(1), 25-33.

Natesan K, Isloor S, Vinayagamurthy B, Ramakrishnaiah S, Doddamane R, & Fooks AR. Developments in Rabies Vaccines: The Path Traversed from Pasteur to the Modern Era of Immunization. Vaccines. 2023; 11(4), 756. DOI:10.3390/vaccines11040756

Lye PY, Kotani E, & Liew MW. Progress and challenges in production of recombinant Newcastle disease virus hemagglutinin-neuraminidase subunit vaccine. Process Biochemistry. 2023. DOI:10.1016/j.procbio.2023.07.023

Hotez PJ, Adhikari R, Chen WH, Chen YL, Gillespie P, Islam NY, Bottazzi ME. From concept to delivery: a yeast-expressed recombinant protein-based COVID-19 vaccine technology suitable for global access. Expert Review of Vaccines. 2023; 22(1), 495-500. DOI:10.1080/14760584.2023.2217917

Kovalenko A, Ryabchevskaya E, Evtushenko E, Nikitin N, & Karpova O. Recombinant Protein Vaccines against Human Betacoronaviruses: Strategies, Approaches and Progress. International Journal of Molecular Sciences. 2023; 24(2), 1701. DOI:10.3390/ijms24021701

Krieg RC, Dong Y, Schwamborn K, & Knuechel R. Protein quantification and its tolerance for different interfering reagents using the BCA-method with regard to 2D SDS PAGE. Journal of biochemical and biophysical methods. 2005; 65(1), 13-19. DOI:10.1016/j.jbbm.2005.08.005

Zhang Z, Yao F, Lv J, Ding Y, Liu X, Zhang L, Pan L. Identification of B-cell epitopes on structural proteins VP1 and VP2 of Senecavirus A and development of a multi-epitope recombinant protein vaccine. Virology. 2023; 582, 48-56. DOI:10.1016/j.virol.2023.03.015

Lin AV. Indirect Elisa. ELISA: methods and protocols. 2015; 51-59.

Redfield RR, Birx DL, Ketter N, Tramont E, Polonis V, Davis C, Burke DS. A phase I evaluation of the safety and immunogenicity of vaccination with recombinant gp160 in patients with early human immunodeficiency virus infection. New England Journal of Medicine. 1991; 324(24), 1677-1684. DOI: 10.1056/NEJM199106133242401

Choi G, Hofstra JJ, Roelofs JJ, Florquin S, Bresser P, Levi M, van der Poll T, & Schultz MJ. Recombinant human activated protein C inhibits local and systemic activation of coagulation without influencing inflammation during Pseudomonas aeruginosa pneumonia in rats. Critical care medicine journal. 2007; 35(5), 1362-1368. DOI:10.1097/01.CCM.0000261888.32654.6D

Foley, H. D., McGettigan, J. P., Siler, C. A., Dietzschold, B., & Schnell, M. J. (2000). A recombinant rabies virus expressing vesicular stomatitis virus glycoprotein fails to protect against rabies virus infection. Proceedings of the National Academy of Sciences, 97(26), 14680-14685. DOI: 10.1073/pnas.011510698

Stewart L, Batesa DO, & Harper SJ. Recombinant human VEGF165b protein is an effective anti-cancer agent in mice. European Journal of Cancer. 2008 ; 44(13), 1883-1894. DOI: 10.1016/j.ejca.2008.05.027

Vermout S, Denis M, Losson B, Mignon B. Choix d'un adjuvant lors d'essais de vaccination. Ann Méd Vét. 2003; 147, 393-401.




How to Cite

Gasmi, M., Rahma, Z., Williams-Hooker, R. W.-H., & Sharma, A. (2023). Production and Characterization of Recombinant Multi-Epitope G Glycoprotein of Rabies Virus. Medical and Pharmaceutical Journal, 2(3), 134–148.



Original Articles


Most read articles by the same author(s)