Refolded dengue virus type 2 NS1 protein expressed in Escherichia coli preserves structural and immunological properties of the native protein.
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.
Amorim
Jaime Henrique
JH
Department of Microbiology, University of São Paulo, Brazil.
Porchia
Bruna F M M
BF
Balan
Andrea
A
Cavalcante
Rafael C M
RC
da Costa
Simone Morais
SM
de Barcelos Alves
Ada Maria
AM
de Souza Ferreira
Luís Carlos
LC
D000914:Antibodies, Viral / Q000276:immunology; D003716:Dengue Virus / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D017510:Protein Folding; D055503:Protein Multimerization; D055550:Protein Stability; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D013057:Spectrum Analysis; D013696:Temperature; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; C079340:NS1 protein, Dengue virus type 2; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins