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Engineering, production, and immunogenicity studies of a truncated form of rabies virus glycoprotein produced in Nicotiana benthamiana plant

Gulshan Mammadova, Irem Gurbuzaslan, Damla Yuksel, Nilufer Gun, Nedim Mutlu, Gulnara Hasanova, Tarlan Mamedov.




Abstract

Rabies is a viral disease caused by the rabies virus (Lyssavirus) that affects the central nervous system, ultimately leading to death and brain disease. Rabies usually transmitted through a bite among domestic dogs and wild carnivorous animals. The rabies virus (RABV) infects the central nervous system causing disease in the brain and death. Rabies is 100% fatal if untreated and ultimately causing more than 70,000 deaths annually worldwide, especially among children in developing countries. Rabies can be prevented by vaccination and can be cured immediately after infection. Although several human and animal vaccines against rabies are available, they are expensive, have relatively low immunogenicity and also difficult to produce. Therefore, less expensive, more immunogenic and safe rabies vaccines that can be produced in the large quantities are urgently needed. The transient plant expression system has become a more attractive and promising platform for the expression of a wide range of recombinant proteins such as vaccines, antibodies, therapeutic proteins, including mammalian complex proteins. In this study, we engineered and produced a truncated form of glycoprotein (G protein) of rabies virus in Nicotiana benthamiana (N. benthamiana) plant for the first time, using transient expression system. Plant produced RG2 protein was purified through Ni-NTA column chromatography. The purification yield of RG2 protein was ~32 mg/ kg plant leaf. The results of immunogenicity studies showed that the plant produced G-protein elicited significantly high antibody titers in mice. Plant-produced G-protein could be a cost-effective, safe and highly immunogenic rabies vaccine candidate.

Key words: Rabies virus, G protein, subunits vaccine, plant transient expression system






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