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

作者：Ho Chin Woi; Tan Wen Siang; Chong Fui Chin; Ling Tau Chuan; Tey Beng Ti^*

来源：Journal of Microbiology and Biotechnology, 2009, 19(4): 416-423.

DOI：10.4014/jmb.0804.254

• Hepatitis B core antigen
• Escherichia coli
• online capture process
• expanded bed adsorption chromatography
• size-exclusion chromatography

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.

• 出版日期2009-4