Expression and Purification of a Novel Computationally Designed Antigen for Simultaneously Detection of HTLV-1 and HBV Antibodies
Computational tools are reliable alternatives to laborious work in chimeric protein design. In this study, a chimeric antigen was designed using computational techniques for simultaneous detection of anti-HTLV-I and anti-HBV in infected sera. Databases were searched for amino acid sequences of HBV/H...
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| Format: | Article |
| Language: | English |
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Tehran University of Medical Sciences
2015-04-01
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| Series: | Iranian Journal of Allergy, Asthma and Immunology |
| Subjects: | |
| Online Access: | https://ijaai.tums.ac.ir/index.php/ijaai/article/view/397 |
| _version_ | 1818110462405902336 |
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| author | Hafez Heydari Zarnagh Mehrdad Ravanshad Ali Akbar Pourfatollah Mohammad Javad Rasaee |
| author_facet | Hafez Heydari Zarnagh Mehrdad Ravanshad Ali Akbar Pourfatollah Mohammad Javad Rasaee |
| author_sort | Hafez Heydari Zarnagh |
| collection | DOAJ |
| description | Computational tools are reliable alternatives to laborious work in chimeric protein design. In this study, a chimeric antigen was designed using computational techniques for simultaneous detection of anti-HTLV-I and anti-HBV in infected sera. Databases were searched for amino acid sequences of HBV/HLV-I diagnostic antigens. The immunodominant fragments were selected based on propensity scales. The diagnostic antigen was designed using these fragments. Secondary and tertiary structures were predicted and the B-cell epitopes were mapped on the surface of built model. The synthetic DNA coding antigen was sub-cloned into pGS21a expression vector. SDS-PAGE analysis showed that glutathione fused antigen was highly expressed in E. coli BL21 (DE3) cells. The recombinant antigen was purified by nickel affinity chromatography. ELISA results showed that soluble antigen could specifically react with the HTLV-I and HBV infected sera. This specific antigen could be used as suitable agent for antibody-antigen based screening tests and can help clinicians in order to perform quick and precise screening of the HBV and HTLV-I infections. |
| first_indexed | 2024-12-11T02:47:32Z |
| format | Article |
| id | doaj.art-8d59305743c54a2cb05f5214acd5fdf2 |
| institution | Directory Open Access Journal |
| issn | 1735-1502 1735-5249 |
| language | English |
| last_indexed | 2024-12-11T02:47:32Z |
| publishDate | 2015-04-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Allergy, Asthma and Immunology |
| spelling | doaj.art-8d59305743c54a2cb05f5214acd5fdf22022-12-22T01:23:23ZengTehran University of Medical SciencesIranian Journal of Allergy, Asthma and Immunology1735-15021735-52492015-04-01142362Expression and Purification of a Novel Computationally Designed Antigen for Simultaneously Detection of HTLV-1 and HBV AntibodiesHafez Heydari Zarnagh0Mehrdad Ravanshad1Ali Akbar Pourfatollah2Mohammad Javad Rasaee3Department of Clinical Biochemistry, Faculity of Medical Science, Tarbiat Modares University, Tehran, Iran.Department of Virology, Faculty of Medicine, Tarbiat Modares University, Tehran, Iran.Department of Immunology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran.Department of Biotechnology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran.Computational tools are reliable alternatives to laborious work in chimeric protein design. In this study, a chimeric antigen was designed using computational techniques for simultaneous detection of anti-HTLV-I and anti-HBV in infected sera. Databases were searched for amino acid sequences of HBV/HLV-I diagnostic antigens. The immunodominant fragments were selected based on propensity scales. The diagnostic antigen was designed using these fragments. Secondary and tertiary structures were predicted and the B-cell epitopes were mapped on the surface of built model. The synthetic DNA coding antigen was sub-cloned into pGS21a expression vector. SDS-PAGE analysis showed that glutathione fused antigen was highly expressed in E. coli BL21 (DE3) cells. The recombinant antigen was purified by nickel affinity chromatography. ELISA results showed that soluble antigen could specifically react with the HTLV-I and HBV infected sera. This specific antigen could be used as suitable agent for antibody-antigen based screening tests and can help clinicians in order to perform quick and precise screening of the HBV and HTLV-I infections.https://ijaai.tums.ac.ir/index.php/ijaai/article/view/397Blood screeningComputational analysisHBVHTLV-ITransfusion |
| spellingShingle | Hafez Heydari Zarnagh Mehrdad Ravanshad Ali Akbar Pourfatollah Mohammad Javad Rasaee Expression and Purification of a Novel Computationally Designed Antigen for Simultaneously Detection of HTLV-1 and HBV Antibodies Iranian Journal of Allergy, Asthma and Immunology Blood screening Computational analysis HBV HTLV-I Transfusion |
| title | Expression and Purification of a Novel Computationally Designed Antigen for Simultaneously Detection of HTLV-1 and HBV Antibodies |
| title_full | Expression and Purification of a Novel Computationally Designed Antigen for Simultaneously Detection of HTLV-1 and HBV Antibodies |
| title_fullStr | Expression and Purification of a Novel Computationally Designed Antigen for Simultaneously Detection of HTLV-1 and HBV Antibodies |
| title_full_unstemmed | Expression and Purification of a Novel Computationally Designed Antigen for Simultaneously Detection of HTLV-1 and HBV Antibodies |
| title_short | Expression and Purification of a Novel Computationally Designed Antigen for Simultaneously Detection of HTLV-1 and HBV Antibodies |
| title_sort | expression and purification of a novel computationally designed antigen for simultaneously detection of htlv 1 and hbv antibodies |
| topic | Blood screening Computational analysis HBV HTLV-I Transfusion |
| url | https://ijaai.tums.ac.ir/index.php/ijaai/article/view/397 |
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