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Refolded dengue virus type 2 NS1 protein expressed in Escherichia coli preserves structural and immunological properties of the native protein.
J Virol Methods. 2010 Aug; 167(2):186-92.JV

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.

Authors+Show Affiliations

Department of Microbiology, University of São Paulo, Brazil.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20399232

Citation

Amorim, Jaime Henrique, et al. "Refolded Dengue Virus Type 2 NS1 Protein Expressed in Escherichia Coli Preserves Structural and Immunological Properties of the Native Protein." Journal of Virological Methods, vol. 167, no. 2, 2010, pp. 186-92.
Amorim JH, Porchia BF, Balan A, et al. Refolded dengue virus type 2 NS1 protein expressed in Escherichia coli preserves structural and immunological properties of the native protein. J Virol Methods. 2010;167(2):186-92.
Amorim, J. H., Porchia, B. F., Balan, A., Cavalcante, R. C., da Costa, S. M., de Barcelos Alves, A. M., & de Souza Ferreira, L. C. (2010). Refolded dengue virus type 2 NS1 protein expressed in Escherichia coli preserves structural and immunological properties of the native protein. Journal of Virological Methods, 167(2), 186-92. https://doi.org/10.1016/j.jviromet.2010.04.003
Amorim JH, et al. Refolded Dengue Virus Type 2 NS1 Protein Expressed in Escherichia Coli Preserves Structural and Immunological Properties of the Native Protein. J Virol Methods. 2010;167(2):186-92. PubMed PMID: 20399232.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Refolded dengue virus type 2 NS1 protein expressed in Escherichia coli preserves structural and immunological properties of the native protein. AU - Amorim,Jaime Henrique, AU - Porchia,Bruna F M M, AU - Balan,Andrea, AU - Cavalcante,Rafael C M, AU - da Costa,Simone Morais, AU - de Barcelos Alves,Ada Maria, AU - de Souza Ferreira,Luís Carlos, Y1 - 2010/04/23/ PY - 2009/09/14/received PY - 2010/03/29/revised PY - 2010/04/08/accepted PY - 2010/4/20/entrez PY - 2010/4/20/pubmed PY - 2010/11/13/medline SP - 186 EP - 92 JF - Journal of virological methods JO - J Virol Methods VL - 167 IS - 2 N2 - 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. SN - 1879-0984 UR - https://www.unboundmedicine.com/medline/citation/20399232/Refolded_dengue_virus_type_2_NS1_protein_expressed_in_Escherichia_coli_preserves_structural_and_immunological_properties_of_the_native_protein_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0166-0934(10)00132-1 DB - PRIME DP - Unbound Medicine ER -