congresos y reuniones científicas
Transient expression of dengue virus envelope protein in a plant system
Rimini, Italia
Congreso; 14th International Biotechnology Symposium and Exhibition; 2010
Introduction. The utilization of plants as bioreactors presents several advantages related with its simplicity, biosafety and low cost. It retains the advantages of eukaryotic expression systems, as post-translational modifications, and prokaryotic expression systems, as scalability and economical production. However, long time-scale and low yield of recombinant proteins have been the main bottlenecks in plant-made biopharmaceuticals. A developed technique known as Magnifection can offset these disadvantages. This transient expression technology is based on replication of viral vectors delivered to the plant by Agrobacterium and allows very fast production, high recombinant protein expression levels. Dengue virus (DV) serotype 2 envelope glycoprotein (E) is the main protein associated with viral entry and the induction of immunity. In order to produce a candidate for subunit vaccines and to provide an antigen for diagnostic kits, E protein was expressed using this technology. Methods. An E truncated version was designed to be expressed alone and co-expressed with DV structural proteins. As well, the critical Domain III of E protein was fused to Hepatitis B core antigen. Nicotiana benthamiana plants were co-infiltrated with 5’ Module, Integrase Module and three different 3’ Module carrying coding sequences for Et, CMEt and HBcore-DV2d3 proteins respectively. Results. The recombinant proteins were produced successfully in N. benthamiana plants and were reactive with anti-E polyclonal antibody and the fusion was reactive with anti-E polyclonal and anti-HBcore antibodies. Discussion. We have demonstrated that Magnifection system is suitable for the production of the recombinant DV-2 protein in N. benthamiana plants. This is the first report of a plant system being able to successfully produce the flaviviral DV-2 E truncated version and its co-expression with C and prM structural DV-2 proteins. Moreover, this is the first work reported in plants in which the HBcore antigen is co-expressed with the critical Domain III of DV-2 Envelope glycoprotein.