Amino acid derivatized monoliths for purification of a DNA vaccine against influenza

Abstract

In recent years, researchers have made several efforts to fight highly contagious respiratory disease caused by influenza virus. One of the best options for reducing the impact of this virus infection is DNA vaccination, therefore it is necessary a large quantity of highly pure plasmid DNA (pDNA) [1, 2]. Thus, in this work it is proposed the production and purification of pDNA expressing influenza virus hemaglutinin protein. For the purification strategy, monoliths are chosen because of their high binding capacity and the excellent mass transfer properties. Agmatine was the ligand of choice, once this amino acid derivative showed to be successful not only in the purification of sc pDNA isoforms but also from complex lysates. The results showed that agmatine is a multifaceted ligand to purify the sc pDNA influenza vaccine under the requirements of the regulatory agencies. In vitro experiments revealed that sc pDNA was able to transfect fibroblast cells and to produce hemaglutinin protein, as proved by immunochemistry analysis with mouse monoclonal anti HA H5N1 IgG primary antibody. The effect of plasmid transfection on cell viability was over 90% as demonstrated with activity of lactate dehydrogenase resazurin assays. In conclusion, our collective approach provides a valuable choice for the efficient isolation of sc pDNA hemaglutinin vaccine which can in near future prevent influenza infection.