Cloning, Expression and Characterization of Hepatitis B Large Surface Antigen In Yeast System
Hepatitis B virus (HBV) infection is a worldwide health problem, which can lead to severe liver disease mainly hepatocellular carcinoma and cirrhosis. Hepatitis B surface antigen (HBsAg) is a glycoprotein found on the surface of the virus and its presence in the blood is an indicator of positive hep...
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| Format: | Thesis |
| Language: | English English |
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2010
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| Online Access: | http://psasir.upm.edu.my/id/eprint/21265/ http://psasir.upm.edu.my/id/eprint/21265/1/FBSB_2010_3_R.pdf |
| _version_ | 1848844174652080128 |
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| author | Mohamad Sinang, Fazia |
| author_facet | Mohamad Sinang, Fazia |
| author_sort | Mohamad Sinang, Fazia |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Hepatitis B virus (HBV) infection is a worldwide health problem, which can lead to severe liver disease mainly hepatocellular carcinoma and cirrhosis. Hepatitis B surface antigen (HBsAg) is a glycoprotein found on the surface of the virus and its presence in the blood is an indicator of positive hepatitis B infection. The existing commercially available vaccine is based on the produced small HBsAg (S-HBsAg) via recombinant DNA technology. Although it is effective, about 5-10% of normal individuals demonstrates no or low response to the standard vaccination schedule. The efficacy of the large HBsAg (L-HBsAg) derived vaccine may differ from S-HBsAg, but such study needs to be conducted to evaluate the potential of L-HBsAg. To date, there is no infonnation on the expression of L-HBsAg intracellularly. Therefore, this study was carried out to express the L-HBsAg protein in yeast Pichia pastoris (P. pastoris) intracellularly. The L-HBsAg gene is inserted into the pPICZ C plasmid and introduced into P. pastoris. The positive clones were confinned by DNA sequencing. Small scale expression of the recombinant L-HBsAg was analyzed by using Western blotting. The glycosylated L-HBsAg of about 42 kDa was detected by Western blotting and its concentration increase from 24 hours to 72 hours sample. The production of L-HBsAg was then scaled up and purified by using sucrose density gradient centrifugation. ELISA showed that the purified L-HBsAg was antigenic. Electron microscopic analysis revealed that the L-HBsAg assembled into spherical particles about 25 nm. The LHBsAg produced in P. pastoris provides an alternative to the vaccines available in market |
| first_indexed | 2025-11-15T08:26:44Z |
| format | Thesis |
| id | upm-21265 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T08:26:44Z |
| publishDate | 2010 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-212652013-05-27T08:16:01Z http://psasir.upm.edu.my/id/eprint/21265/ Cloning, Expression and Characterization of Hepatitis B Large Surface Antigen In Yeast System Mohamad Sinang, Fazia Hepatitis B virus (HBV) infection is a worldwide health problem, which can lead to severe liver disease mainly hepatocellular carcinoma and cirrhosis. Hepatitis B surface antigen (HBsAg) is a glycoprotein found on the surface of the virus and its presence in the blood is an indicator of positive hepatitis B infection. The existing commercially available vaccine is based on the produced small HBsAg (S-HBsAg) via recombinant DNA technology. Although it is effective, about 5-10% of normal individuals demonstrates no or low response to the standard vaccination schedule. The efficacy of the large HBsAg (L-HBsAg) derived vaccine may differ from S-HBsAg, but such study needs to be conducted to evaluate the potential of L-HBsAg. To date, there is no infonnation on the expression of L-HBsAg intracellularly. Therefore, this study was carried out to express the L-HBsAg protein in yeast Pichia pastoris (P. pastoris) intracellularly. The L-HBsAg gene is inserted into the pPICZ C plasmid and introduced into P. pastoris. The positive clones were confinned by DNA sequencing. Small scale expression of the recombinant L-HBsAg was analyzed by using Western blotting. The glycosylated L-HBsAg of about 42 kDa was detected by Western blotting and its concentration increase from 24 hours to 72 hours sample. The production of L-HBsAg was then scaled up and purified by using sucrose density gradient centrifugation. ELISA showed that the purified L-HBsAg was antigenic. Electron microscopic analysis revealed that the L-HBsAg assembled into spherical particles about 25 nm. The LHBsAg produced in P. pastoris provides an alternative to the vaccines available in market 2010-08 Thesis NonPeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/21265/1/FBSB_2010_3_R.pdf Mohamad Sinang, Fazia (2010) Cloning, Expression and Characterization of Hepatitis B Large Surface Antigen In Yeast System. PhD thesis, Universiti Putra Malaysia. Antigens Yeast Liver - Diseases English |
| spellingShingle | Antigens Yeast Liver - Diseases Mohamad Sinang, Fazia Cloning, Expression and Characterization of Hepatitis B Large Surface Antigen In Yeast System |
| title | Cloning, Expression and Characterization of Hepatitis B Large Surface Antigen In Yeast System |
| title_full | Cloning, Expression and Characterization of Hepatitis B Large Surface Antigen In Yeast System |
| title_fullStr | Cloning, Expression and Characterization of Hepatitis B Large Surface Antigen In Yeast System |
| title_full_unstemmed | Cloning, Expression and Characterization of Hepatitis B Large Surface Antigen In Yeast System |
| title_short | Cloning, Expression and Characterization of Hepatitis B Large Surface Antigen In Yeast System |
| title_sort | cloning, expression and characterization of hepatitis b large surface antigen in yeast system |
| topic | Antigens Yeast Liver - Diseases |
| url | http://psasir.upm.edu.my/id/eprint/21265/ http://psasir.upm.edu.my/id/eprint/21265/1/FBSB_2010_3_R.pdf |