Optimizations of expression and purification of recombinant HIV-1 CRF01_AE p24 protein in Escherichia coli for development of immunodiagnostic assay
<p><strong>Background:</strong> Conventional method for confirmation of HIV infection is Western blot. However, it has limitations because of contamination by human cellular antigen and genetic diversity among the HIV-1 subtypes that show indeterminate result and inaccuracy for the...
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Faculty of Medicine Universitas Indonesia
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doaj-e80a39e2cf3347cbaf23302bf40588192020-11-24T22:08:41ZengFaculty of Medicine Universitas Indonesia Medical Journal of Indonesia0853-17732252-80832015-04-0124114810.13181/mji.v24i1.11661068Optimizations of expression and purification of recombinant HIV-1 CRF01_AE p24 protein in Escherichia coli for development of immunodiagnostic assayAde P.R. Simaremare0Budiman Bela1Andi Yasmon2Fera Ibrahim3Biomedical Programme, Faculty of Medicine, Universitas Indonesia, JakartaInstitute of Human Virology and Cancer Biology, Faculty of Medicine, Universitas Indonesia, JakartaDepartment of Microbiology, Faculty of Medicine, Universitas Indonesia, JakartaDepartment of Microbiology, Faculty of Medicine, Universitas Indonesia, Jakarta<p><strong>Background:</strong> Conventional method for confirmation of HIV infection is Western blot. However, it has limitations because of contamination by human cellular antigen and genetic diversity among the HIV-1 subtypes that show indeterminate result and inaccuracy for the diagnosis of different strains. Most of Western blot developed are based on HIV-1 B subtype. In Indonesia HIV-1 CRF01_AE subtype is dominantly circulated. Therefore, we optimized the expression, purification of the recombinant HIV-1 CRF01_AE p24 protein for development of immunodiagnostic assay.</p><p><strong>Methods:</strong> Optimization of protein expression in Escherichia coli strain BL21CP was performed including induction time, isopropyl-1-thio-d-galactopyranoside (IPTG) and immidazole consentrations, and induction temperature. Purification of the recombinant p24 protein was used by using Ni-NTA (Qiagen) purification system in native condition. </p><p><strong>Results:</strong> Expression and purification of HIV-1 CRF01_AE p24 protein have been performed. Confirmation of the recombinant protein by Western blot showed the expression and purification of recombinant p24 protein has been optimized well and reactive with sera of patients with HIV-1 CRF01_AE subtype positive.</p><p><strong>Conclusion:</strong> The recombinant HIV-1 CRF01_AE p24 protein has been expressed and purified successfully, and it is potential to be used as antigen for immunodiagnostic assay.</p>http://mji.ui.ac.id/journal/index.php/mji/article/view/1166expressionHIV-1 CRF01_AEp24 proteinpurificationrecombinant protein |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ade P.R. Simaremare Budiman Bela Andi Yasmon Fera Ibrahim |
spellingShingle |
Ade P.R. Simaremare Budiman Bela Andi Yasmon Fera Ibrahim Optimizations of expression and purification of recombinant HIV-1 CRF01_AE p24 protein in Escherichia coli for development of immunodiagnostic assay Medical Journal of Indonesia expression HIV-1 CRF01_AE p24 protein purification recombinant protein |
author_facet |
Ade P.R. Simaremare Budiman Bela Andi Yasmon Fera Ibrahim |
author_sort |
Ade P.R. Simaremare |
title |
Optimizations of expression and purification of recombinant HIV-1 CRF01_AE p24 protein in Escherichia coli for development of immunodiagnostic assay |
title_short |
Optimizations of expression and purification of recombinant HIV-1 CRF01_AE p24 protein in Escherichia coli for development of immunodiagnostic assay |
title_full |
Optimizations of expression and purification of recombinant HIV-1 CRF01_AE p24 protein in Escherichia coli for development of immunodiagnostic assay |
title_fullStr |
Optimizations of expression and purification of recombinant HIV-1 CRF01_AE p24 protein in Escherichia coli for development of immunodiagnostic assay |
title_full_unstemmed |
Optimizations of expression and purification of recombinant HIV-1 CRF01_AE p24 protein in Escherichia coli for development of immunodiagnostic assay |
title_sort |
optimizations of expression and purification of recombinant hiv-1 crf01_ae p24 protein in escherichia coli for development of immunodiagnostic assay |
publisher |
Faculty of Medicine Universitas Indonesia |
series |
Medical Journal of Indonesia |
issn |
0853-1773 2252-8083 |
publishDate |
2015-04-01 |
description |
<p><strong>Background:</strong> Conventional method for confirmation of HIV infection is Western blot. However, it has limitations because of contamination by human cellular antigen and genetic diversity among the HIV-1 subtypes that show indeterminate result and inaccuracy for the diagnosis of different strains. Most of Western blot developed are based on HIV-1 B subtype. In Indonesia HIV-1 CRF01_AE subtype is dominantly circulated. Therefore, we optimized the expression, purification of the recombinant HIV-1 CRF01_AE p24 protein for development of immunodiagnostic assay.</p><p><strong>Methods:</strong> Optimization of protein expression in Escherichia coli strain BL21CP was performed including induction time, isopropyl-1-thio-d-galactopyranoside (IPTG) and immidazole consentrations, and induction temperature. Purification of the recombinant p24 protein was used by using Ni-NTA (Qiagen) purification system in native condition. </p><p><strong>Results:</strong> Expression and purification of HIV-1 CRF01_AE p24 protein have been performed. Confirmation of the recombinant protein by Western blot showed the expression and purification of recombinant p24 protein has been optimized well and reactive with sera of patients with HIV-1 CRF01_AE subtype positive.</p><p><strong>Conclusion:</strong> The recombinant HIV-1 CRF01_AE p24 protein has been expressed and purified successfully, and it is potential to be used as antigen for immunodiagnostic assay.</p> |
topic |
expression HIV-1 CRF01_AE p24 protein purification recombinant protein |
url |
http://mji.ui.ac.id/journal/index.php/mji/article/view/1166 |
work_keys_str_mv |
AT adeprsimaremare optimizationsofexpressionandpurificationofrecombinanthiv1crf01aep24proteininescherichiacolifordevelopmentofimmunodiagnosticassay AT budimanbela optimizationsofexpressionandpurificationofrecombinanthiv1crf01aep24proteininescherichiacolifordevelopmentofimmunodiagnosticassay AT andiyasmon optimizationsofexpressionandpurificationofrecombinanthiv1crf01aep24proteininescherichiacolifordevelopmentofimmunodiagnosticassay AT feraibrahim optimizationsofexpressionandpurificationofrecombinanthiv1crf01aep24proteininescherichiacolifordevelopmentofimmunodiagnosticassay |
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