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Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis.
J Virol Methods. 2011 Jul; 175(1):109-16.JV

Abstract

The non-structural 1 (NS1) protein plays an important role in dengue diagnosis because it has been detected as a soluble serum antigen in both primary and secondary infections. The NS1 protein was expressed in Escherichia coli cells, and the efficiency of four different refolding protocols was tested. All of the protocols generated dimeric NS1 in a conformation similar to that of the protein expressed by eukaryotic cells. A polyclonal antibody produced from the properly folded E. coli recombinant NS1 (rNS1) protein proved to be a useful tool for the diagnosis of Dengue virus because it detected 100% of the Dengue virus 2 (DENV2) in infected patients' sera and 60% of the DENV IgM-positive sera not detected by commercial NS1-based diagnostic kits. These data suggest a high-efficiency method for correctly folding rNS1 that maintains its structural and immunogenic properties. In addition, a detection method using the polyclonal antibody against correctly folded rNS1 seemed to be more sensitive and efficient for NS1 detection in serum, highlighting its usefulness for developing a high-sensitivity diagnostic kit.

Authors+Show Affiliations

Laboratório de Genômica Estrutural, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-590, Brazil.No affiliation info availableNo affiliation info availableNo 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

21569796

Citation

Allonso, Diego, et al. "Polyclonal Antibodies Against Properly Folded Dengue Virus NS1 Protein Expressed in E. Coli Enable Sensitive and Early Dengue Diagnosis." Journal of Virological Methods, vol. 175, no. 1, 2011, pp. 109-16.
Allonso D, da Silva Rosa M, Coelho DR, et al. Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis. J Virol Methods. 2011;175(1):109-16.
Allonso, D., da Silva Rosa, M., Coelho, D. R., da Costa, S. M., Nogueira, R. M., Bozza, F. A., Santos, F. B., de Barcelos Alves, A. M., & Mohana-Borges, R. (2011). Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis. Journal of Virological Methods, 175(1), 109-16. https://doi.org/10.1016/j.jviromet.2011.04.029
Allonso D, et al. Polyclonal Antibodies Against Properly Folded Dengue Virus NS1 Protein Expressed in E. Coli Enable Sensitive and Early Dengue Diagnosis. J Virol Methods. 2011;175(1):109-16. PubMed PMID: 21569796.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis. AU - Allonso,Diego, AU - da Silva Rosa,Marcela, AU - Coelho,Diego Rodrigues, AU - da Costa,Simone Morais, AU - Nogueira,Rita Maria Ribeiro, AU - Bozza,Fernando Augusto, AU - Santos,Flavia Barreto Dos, AU - de Barcelos Alves,Ada Maria, AU - Mohana-Borges,Ronaldo, Y1 - 2011/05/04/ PY - 2010/12/14/received PY - 2011/04/25/revised PY - 2011/04/27/accepted PY - 2011/5/17/entrez PY - 2011/5/17/pubmed PY - 2011/12/13/medline SP - 109 EP - 16 JF - Journal of virological methods JO - J Virol Methods VL - 175 IS - 1 N2 - The non-structural 1 (NS1) protein plays an important role in dengue diagnosis because it has been detected as a soluble serum antigen in both primary and secondary infections. The NS1 protein was expressed in Escherichia coli cells, and the efficiency of four different refolding protocols was tested. All of the protocols generated dimeric NS1 in a conformation similar to that of the protein expressed by eukaryotic cells. A polyclonal antibody produced from the properly folded E. coli recombinant NS1 (rNS1) protein proved to be a useful tool for the diagnosis of Dengue virus because it detected 100% of the Dengue virus 2 (DENV2) in infected patients' sera and 60% of the DENV IgM-positive sera not detected by commercial NS1-based diagnostic kits. These data suggest a high-efficiency method for correctly folding rNS1 that maintains its structural and immunogenic properties. In addition, a detection method using the polyclonal antibody against correctly folded rNS1 seemed to be more sensitive and efficient for NS1 detection in serum, highlighting its usefulness for developing a high-sensitivity diagnostic kit. SN - 1879-0984 UR - https://www.unboundmedicine.com/medline/citation/21569796/Polyclonal_antibodies_against_properly_folded_Dengue_virus_NS1_protein_expressed_in_E__coli_enable_sensitive_and_early_dengue_diagnosis_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0166-0934(11)00167-4 DB - PRIME DP - Unbound Medicine ER -