دورية أكاديمية
Rational Design and Evaluation of the Recombinant Multiepitope Protein for Serodiagnosis of Rubella.
| العنوان: | Rational Design and Evaluation of the Recombinant Multiepitope Protein for Serodiagnosis of Rubella. |
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| المؤلفون: | Souza M; Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Departamento de Biologia Celular, Universidade de Brasília, Brasília, 70910-900, Brasília, DF, Brasil., Machado J; Laboratório de Biotecnologia de Microrganismos, Universidade Federal de São João Del-Rei, Campus Centro-Oeste Dona Lindu, 35501-296, Divinópolis, MG, Brasil., da Silva J; Laboratório de Biotecnologia de Microrganismos, Universidade Federal de São João Del-Rei, Campus Centro-Oeste Dona Lindu, 35501-296, Divinópolis, MG, Brasil., Ramos L; Laboratório de Biotecnologia de Microrganismos, Universidade Federal de São João Del-Rei, Campus Centro-Oeste Dona Lindu, 35501-296, Divinópolis, MG, Brasil., Nogueira L; Laboratório de Biotecnologia de Microrganismos, Universidade Federal de São João Del-Rei, Campus Centro-Oeste Dona Lindu, 35501-296, Divinópolis, MG, Brasil., Ribeiro P; Laboratório de Biotecnologia de Microrganismos, Universidade Federal de São João Del-Rei, Campus Centro-Oeste Dona Lindu, 35501-296, Divinópolis, MG, Brasil., Dias D; Laboratório de Biotecnologia de Microrganismos, Universidade Federal de São João Del-Rei, Campus Centro-Oeste Dona Lindu, 35501-296, Divinópolis, MG, Brasil., Santos J; Laboratório de Biotecnologia de Microrganismos, Universidade Federal de São João Del-Rei, Campus Centro-Oeste Dona Lindu, 35501-296, Divinópolis, MG, Brasil., Santos JC; Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Departamento de Biologia Celular, Universidade de Brasília, Brasília, 70910-900, Brasília, DF, Brasil., Nóbrega Y; Laboratório de Doenças Imunogenéticase Crônico-degenerativas, Faculdade de Saúde, Departamento de Ciências Farmacêuticas, Universidade de Brasília, Brasília, DF, 70910-900, Brasil., Souza A; Laboratório de Biofísica, Instituto de Ciências Biológicas, Departamento de Biologia Celular, Universidade de Brasília, Brasília, DF, 70910-900, Brasil., Freitas S; Laboratório de Biofísica, Instituto de Ciências Biológicas, Departamento de Biologia Celular, Universidade de Brasília, Brasília, DF, 70910-900, Brasil., da Paz MC; Laboratório de Nanobiotecnologia and Biativos, Universidade Federal de São João Del-Rei, Campus Centro-Oeste Dona Lindu, Divinópolis, MG, 35501-296, Brasil., Felipe M; Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Departamento de Biologia Celular, Universidade de Brasília, Brasília, 70910-900, Brasília, DF, Brasil., Torres F; Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Departamento de Biologia Celular, Universidade de Brasília, Brasília, 70910-900, Brasília, DF, Brasil., Galdino A; Laboratório de Biotecnologia de Microrganismos, Universidade Federal de São João Del-Rei, Campus Centro-Oeste Dona Lindu, 35501-296, Divinópolis, MG, Brasil. |
| المصدر: | Current pharmaceutical biotechnology [Curr Pharm Biotechnol] 2022; Vol. 23 (8), pp. 1094-1100. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Bentham Science Publishers Country of Publication: Netherlands NLM ID: 100960530 Publication Model: Print Cited Medium: Internet ISSN: 1873-4316 (Electronic) Linking ISSN: 13892010 NLM ISO Abbreviation: Curr Pharm Biotechnol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Hilversum, Netherlands ; Boca Raton, FL : Bentham Science Publishers, c2000- |
| مواضيع طبية MeSH: | Antibodies, Viral* , Rubella*/diagnosis, Child ; Enzyme-Linked Immunosorbent Assay/methods ; Epitopes/genetics ; Escherichia coli/genetics ; Female ; Humans ; Pregnancy ; Recombinant Proteins/genetics ; Serologic Tests |
| مستخلص: | Background: Rubella is an infection caused by rubella virus (RV) and is generally regarded as a mild childhood disease. The disease continues to be of public health importance mainly because when the infection is acquired during early pregnancy, it often results in fetal abnormalities, which are classified as congenital rubella syndrome (CRS). An accurate diagnosis of rubella is thus of pivotal importance for proper treatment. Objectives: The aim of the study was to produce a recombinant multiepitope protein (rMERUB) for the diagnosis of rubella, based on conserved immunodominant epitopes of glycoprotein E1 and E2. Methods: A synthetic gene was designed and cloned into vector pET21a with a 6xHis tag at the Cterminal for affinity purification and overexpressed in Escherichia coli cells. Biophysical analysis of rMERUB was performed by circular dichroism. Biological activity was assessed using an in-house ELISA assay. Results: Expression in Escherichia coli showed a ~22 kDa protein that was purified and used to perform structural assays and an IgG ELISA. Structural analyses reveal that rMERUB has a β leaf pattern that promotes the exposure of epitopes, thus allowing antibody recognition. Evaluation of 33 samples (22=positive; 11=negative) was performed using in-house ELISA and this was compared with a commercial kit. The sensitivity was 100% (95% CI: 85-100) and specificity 90.91% (95% CI: 62-99). Excellent agreement (Kappa index = 0.9) was obtained between ELISA assays. Conclusion: The careful choice of epitopes and the high epitope density, coupled with simple-step purification, pinpoints rMERUB as a promising alternative for rubella diagnosis, with potential for the development of a diagnostic kit. (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.) |
| معلومات مُعتمدة: | 193.000.582/2009, 193.000.856/2015 Fundação de Amparo à Pesquisa do Distrito Federal (FAP-DF); 563855/2010, 564000/2010-8 309244/2013-7, 315009/2018-7, 428962/2018-1 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq); APQ- 00835/14 Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG); 001 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES |
| فهرسة مساهمة: | Keywords: Rubella virus; diagnostic test; enzyme immunoassay; fetal abnormalities; glycoprotein; multiepitope protein |
| المشرفين على المادة: | 0 (Antibodies, Viral) 0 (Epitopes) 0 (Recombinant Proteins) |
| تواريخ الأحداث: | Date Created: 20210908 Date Completed: 20220517 Latest Revision: 20220517 |
| رمز التحديث: | 20240628 |
| DOI: | 10.2174/1389201022666210907170921 |
| PMID: | 34493182 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1873-4316 |
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| DOI: | 10.2174/1389201022666210907170921 |