Yap, Wei Boon and Tey, Beng Ti and Mohamed 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, 1217 (21). pp. 3473-3480. ISSN 0021-9673
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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).
| Item Type: | Article |
|---|---|
| Keyword: | IMA-EBAC, His-tagged HBcAg, Unclarified bacterial homogenate, Streamline Chelating, Virus-like particles |
| Faculty or Institute: | Faculty of Engineering |
| Publisher: | Elsevier |
| DOI Number: | 10.1016/j.chroma.2010.03.012 |
| ID Code: | 11270 |
| Deposited By: | Anas Yahaya |
| Deposited On: | 29 Mar 2011 17:39 |
| Last Modified: | 29 Mar 2011 17:39 |
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