Refolded dengue virus type 2 NS1 protein expressed in Escherichia coli preserves structural and immunological properties of the native protein

被引：41

作者：

Amorim, Jaime Henrique

Porchia, Bruna F. M. M.

Balan, Andrea ^{[2
]}

Cavalcante, Rafael C. M.

da Costa, Simone Morais ^{[3
]}

de Barcelos Alves, Ada Maria ^{[3
]}

de Souza Ferreira, Luis Carlos ^{[1
]}

机构：

[1] Univ Sao Paulo, Lab Desenvolvimento Vacinas, Dept Microbiol, BR-05508900 Sao Paulo, Brazil

[2] Brazilian Synchrotron Light Lab LNLS, Ctr Struct Mol Biol CeBiME, Campinas, Brazil

[3] Fundacao Oswaldo Cruz, Lab Biotechnol & Physiol Virus Infect, Inst Oswaldo Cruz, Rio De Janeiro, Brazil

来源：

JOURNAL OF VIROLOGICAL METHODS | 2010年 / 167卷 / 02期

基金：

巴西圣保罗研究基金会;

关键词：

Dengue virus; NS1; protein; Protein purification; Vaccines; STRUCTURE PREDICTION; IMMUNIZATION; ENCEPHALITIS; PROTECTION; ANTIBODY; FEVER; MICE;

D O I：

10.1016/j.jviromet.2010.04.003

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (C) 2010 Elsevier B V All rights reserved

引用

页码：186 / 192

页数：7

共 30 条

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