The development of an effective vaccine for the control of dengue fever is a priority for countries where the disease prevails. The dengue virus NS1 protein has been shown to be a protective antigen in experimental mouse models. However, attempts to develop acellular vaccines based on recombinant NS1 protein produced in bacterial expression systems has been hampered by the difficulty to obtain a soluble protein with preserved structural and immunological features with regard to the native protein. In the present study, experimental conditions leading to purification of a recombinant dengue virus type 2 (DENV-2) NS1 protein expressed in Escherichia coli are reported. The refolded recombinant protein remained fully soluble and formed stable dimers with a secondary structure encompassing 41% β -sheets, 14% α -helixes and 45% of turns. Antibodies that predominantly recognized conformational epitopes of the NS1 protein expressed in eukaryotic cells recognized the NS1 protein expressed in E. coli. Collectively, the results demonstrate that the recombinant NS1 protein preserved important conformation and antigenic determinants of the native virus protein and represents a valuable reagent both for development of vaccines and diagnostic tools.
Full conference title:
110th General Meeting American Society for Microbiology
- ASM 110th (2010)