Development of Recombinant Protein-Based Vaccine Against Classical Swine Fever Virus in Pigs Using Transgenic Nicotiana benthamiana.
Classical swine fever virus (CSFV) is highly contagious and fatal to infected pigs. Vaccines against CSFV have been developed from inactivated or modified live virus. The vaccine is effective for immunization of animals, but they are associated with E.coli Recombinant Proteins problems such as accidental spread of animal viruses in the field, and with trade barriers following vaccination. Here, we report the generation of transgenic Nicotiana benthamiana plants for large-scale, cost-effective production E2 fusion proteins for use as a recombinant vaccine against CSFV in pigs. Transgenic N. benthamiana plants save an intergenic, single-copy insertion of a chimeric gene encoding a fusion protein E2 has a high level of expression of the transgene. For large-scale production of the fusion protein E2 leaf tissue, we developed protein purification protocol consisting of a cellulose-binding domain (CBD) -cellulose affinity based purification and size-exclusion gel filtration chromatogr...