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http://dx.doi.org/10.4014/jmb.0804.254

A Preparative Purification Process for Recombinant Hepatitis B Core Antigen Using Online Capture by Expanded Bed Adsorption Followed by Size-Exclusion Chromatography  

Ho, Chin Woi (Department of Chemical and Environmental Engineering, Faculty of Engineering, Universiti Putra Malaysia)
Tan, Wen Siang (Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia)
Chong, Fui Chin (Department of Chemical and Environmental Engineering, Faculty of Engineering, Universiti Putra Malaysia)
Ling, Tau Chuan (Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia)
Tey, Beng Ti (Department of Chemical and Environmental Engineering, Faculty of Engineering, Universiti Putra Malaysia)
Publication Information
Journal of Microbiology and Biotechnology / v.19, no.4, 2009 , pp. 416-423 More about this Journal
Abstract
Hepatitis B core antigen(HBcAg) is an important serological marker used in the diagnosis of hepatitis B virus(HBV) infections. In the current study, a fast and efficient preparative purification protocol for truncated HBcAg from Escherichia coli disruptate was developed. The recombinant HBcAg was first captured by anion exchange expanded bed adsorption chromatography integrated with a cell disruption process. This online capture process has shortened the process time and eliminated the "hold-up" period that may be detrimental to the quality of target protein. The eluted product from the expanded bed adsorption chromatography was subsequently purified using size-exclusion chromatography. The results showed that this novel purification protocol achieved a recovery yield of 45.1% with a product purity of 88.2%, which corresponds to a purification factor of 4.5. The recovered HBcAg is still biologically active as shown by ELISA test.
Keywords
Hepatitis B core antigen; Escherichia coli; online capture process; expanded bed adsorption chromatography; size-exclusion chromatography;
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