Novel BPI3Vc-vectored surface displayed fusion and hemagglutinin-neuraminidase antigens elicit broadly neutralizing antibodies in cattle

Novel BPI3V Fusion and Hemagglutinin-Neuraminidase genes, designated F2 and HN2, respectively, were designed. Briefly, sixty-two BPI3V genotypes A-C sequences encoding the F protein and thirty-five BPI3V genotypes A-C sequences encoding the HN protein were retrieved from NCBI, representing all the BPI3V genotypes A-C F and HN protein sequences that were available. Multiple sequence alignment was conducted using MEGA 7, which generated consensus sequences for both polypeptides based on a > 50 % identity threshold. In cases where identity was less than 50 %, the most dominant amino acid was selected. If there was neither a consensus nor a dominant amino acid, residues from the BPI3Vc were selected. The final consensus sequences for the F2 and HN2 antigens contained conserved polypeptide sequences from the BPI3V genotypes A-C genomes. The polypeptide sequences were used to generate synthetic F2 and HN2 genes that were codon-optimized for expression in Bos taurus (GenScript).