Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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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

  • 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)
  • Published : 2009.04.30
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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.

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References

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