Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography
Hepatitis B core antigen (HBcAg) is used as a diagnostic reagent for the detection of hepatitis B virus infection. In this study, immobilized metal affinity-expanded bed adsorption chromatography (IMA-EBAC) was employed to purify N-terminally His-tagged HBcAg from unclarified bacterial homogenate. S...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2010
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| Online Access: | http://psasir.upm.edu.my/id/eprint/11270/ http://psasir.upm.edu.my/id/eprint/11270/1/Purification%20of%20His-tagged%20hepatitis%20B%20core%20antigen%20from%20unclarified%20bacterial%20homogenate%20using%20immobilized%20metal%20affinity-expanded%20bed%20adsorption%20chromatography.pdf |
| _version_ | 1848841604418240512 |
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| author | Yap, Wei Boon Tey, Beng Ti Mohammed Alitheen, Noorjahan Banu Tan, Wen Siang |
| author_facet | Yap, Wei Boon Tey, Beng Ti Mohammed Alitheen, Noorjahan Banu Tan, Wen Siang |
| author_sort | Yap, Wei Boon |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Hepatitis B core antigen (HBcAg) is used as a diagnostic reagent for the detection of hepatitis B virus infection. In this study, immobilized metal affinity-expanded bed adsorption chromatography (IMA-EBAC) was employed to purify N-terminally His-tagged HBcAg from unclarified bacterial homogenate. Streamline Chelating was used as the adsorbent and the batch adsorption experiment showed that the optimal binding pH of His-tagged HBcAg was 8.0 with a binding capacity of 1.8 mg per ml of adsorbent. The optimal elution condition for the elution of His-tagged HBcAg from the adsorbent was at pH 7 in the presence of 500 mM imidazole and 1.5 M NaCl. The IMA-EBAC has successfully recovered 56% of His-tagged HBcAg from the unclarified E. coli homogenate with a purification factor of 3.64. Enzyme-linked immunosorbent assay (ELISA) showed that the antigenicity of the recovered His-tagged HBcAg was not affected throughout the IMA-EBAC purification process and electron microscopy revealed that the protein assembled into virus-like particles (VLP). |
| first_indexed | 2025-11-15T07:45:53Z |
| format | Article |
| id | upm-11270 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T07:45:53Z |
| publishDate | 2010 |
| publisher | Elsevier |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-112702016-11-30T08:11:41Z http://psasir.upm.edu.my/id/eprint/11270/ Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography Yap, Wei Boon Tey, Beng Ti Mohammed Alitheen, Noorjahan Banu Tan, Wen Siang Hepatitis B core antigen (HBcAg) is used as a diagnostic reagent for the detection of hepatitis B virus infection. In this study, immobilized metal affinity-expanded bed adsorption chromatography (IMA-EBAC) was employed to purify N-terminally His-tagged HBcAg from unclarified bacterial homogenate. Streamline Chelating was used as the adsorbent and the batch adsorption experiment showed that the optimal binding pH of His-tagged HBcAg was 8.0 with a binding capacity of 1.8 mg per ml of adsorbent. The optimal elution condition for the elution of His-tagged HBcAg from the adsorbent was at pH 7 in the presence of 500 mM imidazole and 1.5 M NaCl. The IMA-EBAC has successfully recovered 56% of His-tagged HBcAg from the unclarified E. coli homogenate with a purification factor of 3.64. Enzyme-linked immunosorbent assay (ELISA) showed that the antigenicity of the recovered His-tagged HBcAg was not affected throughout the IMA-EBAC purification process and electron microscopy revealed that the protein assembled into virus-like particles (VLP). Elsevier 2010-05-21 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/11270/1/Purification%20of%20His-tagged%20hepatitis%20B%20core%20antigen%20from%20unclarified%20bacterial%20homogenate%20using%20immobilized%20metal%20affinity-expanded%20bed%20adsorption%20chromatography.pdf Yap, Wei Boon and Tey, Beng Ti and Mohammed Alitheen, Noorjahan Banu and Tan, Wen Siang (2010) Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography. Journal of Chromatography A, 1217 (21). pp. 3473-3480. ISSN 0021-9673; ESSN: 1873-3778 10.1016/j.chroma.2010.03.012 |
| spellingShingle | Yap, Wei Boon Tey, Beng Ti Mohammed Alitheen, Noorjahan Banu Tan, Wen Siang Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography |
| title | Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography |
| title_full | Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography |
| title_fullStr | Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography |
| title_full_unstemmed | Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography |
| title_short | Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography |
| title_sort | purification of his-tagged hepatitis b core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography |
| url | http://psasir.upm.edu.my/id/eprint/11270/ http://psasir.upm.edu.my/id/eprint/11270/ http://psasir.upm.edu.my/id/eprint/11270/1/Purification%20of%20His-tagged%20hepatitis%20B%20core%20antigen%20from%20unclarified%20bacterial%20homogenate%20using%20immobilized%20metal%20affinity-expanded%20bed%20adsorption%20chromatography.pdf |