| Literature DB >> 20399232 |
Jaime Henrique Amorim1, Bruna F M M Porchia, Andrea Balan, Rafael C M Cavalcante, Simone Morais da Costa, Ada Maria de Barcelos Alves, Luís Carlos de Souza Ferreira.
Abstract
The dengue virus NS1 protein has been shown to be a protective antigen under different experimental conditions but the recombinant protein produced in bacterial expression systems is usually not soluble and loses structural and immunological features of the native viral protein. In the present study, experimental conditions leading to purification and refolding of the recombinant dengue virus type 2 (DENV-2) NS1 protein expressed in Escherichia coli are described. The refolded recombinant protein was recovered as heat-stable soluble dimers with preserved structural features, as demonstrated by spectroscopic methods. In addition, antibodies against epitopes of the NS1 protein expressed in eukaryotic cells recognized the refolded protein expressed in E. coli but not the denatured form or the same protein submitted to a different refolding condition. 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 either for the development of vaccines or for diagnostic methods. Copyright (c) 2010 Elsevier B.V. All rights reserved.Entities:
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Year: 2010 PMID： 20399232 DOI： 10.1016/j.jviromet.2010.04.003
Source DB: PubMed Journal: J Virol Methods ISSN： 0166-0934 Impact factor: 2.014