Citation
Abstract
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).
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Additional Metadata
| Item Type: | Article |
|---|---|
| Divisions: | Faculty of Engineering Faculty of Biotechnology and Biomolecular Sciences Institute of Bioscience |
| DOI Number: | https://doi.org/10.1016/j.chroma.2010.03.012 |
| Publisher: | Elsevier |
| Keywords: | IMA-EBAC; His-tagged HBcAg; Unclarified bacterial homogenate; Streamline Chelating; Virus-like particles |
| Depositing User: | Anas Yahaya |
| Date Deposited: | 29 Mar 2011 09:39 |
| Last Modified: | 30 Nov 2016 08:11 |
| Altmetrics: | http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1016/j.chroma.2010.03.012 |
| URI: | http://psasir.upm.edu.my/id/eprint/11270 |
| Statistic Details: | View Download Statistic |
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