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KMID : 0545120090190040416
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Journal of Microbiology and Biotechnology
2009 Volume.19 No. 4 p.416 ~ p.423
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A Preparative Purification Process for Recombinant Hepatitis B Core Antigen Using Online Capture by Expanded Bed Adsorption Followed by Size-Exclusion Chromatography
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Ho Chin-Woi
Tan Wen-Siang
Chong Fui-Chin
Ling Tau-Chuan
Tey Beng-Ti
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Abstract
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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.
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KEYWORD
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Hepatitis B core antigen, Escherichia coli, online capture process, expanded bed adsorption chromatography, size-exclusion chromatography
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