Enhanced immunogenicity of the rabies virus glycoprotein fused to a dendritic cell-targeting peptide.

The dendritic cell-targeting peptide (DCP) trimer sequence has previously shown to enhance specific IgG induction. This study aims to investigate the expression of rabies virus glycoprotein (RVG) in eukaryotic cells through cloning in the pCDNA3.1 + vector, alongside the fusion of a DCP. The DCP trimer sequence was fused to the C-terminal of the RVG to improve the immune response. The construct was designed to include a His-tag for protein purification and an enterokinase (EK) cleavage site for separation of the tag from the recombinant protein. The RVG gene was amplified from the rabies virus genome via RT-PCR, and cloning was performed using BamHI and EcoRI restriction enzymes. The recombinant plasmid (RVG- pCDNA3.1+) was transfected into BHK-21 cells using lipofection, and the expression of the recombinant protein was confirmed through SDS-PAGE and western blotting using anti-His antibodies. The His-tagged recombinant protein was purified using Ni-NTA resin. Immunogenicity of the recombinant protein was investigated through mouse inoculation and analysis of the serum samples using RFFIT and ELISA methods. The results indicated successful cloning and expression of the RVG-DCP fusion protein in eukaryotic cells, with potential applications in rabies vaccine development. The results suggests a novel method for enhancing the immunogenicity of viral glycoproteins through the use of dendritic cell-targeting peptides.