A multi-epitope-based subunit vaccine candidate against peste des petit generated through reverse vaccinology

Abstract


Willis A Adero*, Steven Nyanjom, Martin Kiogora Mwirigi, Yattinder B, Samuel Onyoyo Nguya and Paul Kimutai kirui

Peste des Petits Ruminant Virus (PPRV) is an acute, highly contagious viral disease of small ruminants, endemic to su-Saharan Africa, Asia and the Arabian Peninsula. The disease is a major constraint to food security, causing significant economic losses to subsistence farmers in affected areas. The fusion and Hemagglutinin proteins are highly immunogenic and produced in large quantities in virus-infected cells. This makes them suitable targets for the host’s immune response. This study aimed at developing and evaluating effective multi-epitope vaccine candidate against PPRV developed from two immunogenic proteins Hemagglutinin (H), and Fusion (F), using a bioinformatics approach. In order to construct the subunit vaccine candidates, plasmid pET28a (+) was used as an expression vector, which was later transformed into E. coli DH5-α cells, followed by plasmid extraction, PCR amplification, and DNA sequencing. The white recombinant colony was selected, cultured, induced with 50 μm Isopropyl β-D-1-Thiogalactopyranoside (IPTG), and identified using the SDS-PAGE electrophoresis method. The protein were then extracted from the gel and evaluated for their immunogenicity. Results demonstrated that the constructed vaccine candidate was expressed successfully and had the expected molecular weight of 65 kDa in-vitro and in-vivo testing of the candidate was done to test for immunogenicity and was demonstrated to recognize Peste des Petits Ruminant (PPR) antibodies from vaccinated rabbits and showed a higher antibody titer than the positive control. In conclusion, the immunogenic proteins are potential vaccine candidates for development of a sub-unit vaccine against PPR.

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