The direct recovery of recombinant hepatitis B core antigen from disruptate derived from continuous-flow bead milling

Chin Woi Ho, Wen Siang Tan, Suryani Kamaruddin, Tau Chuan Ling, Beng Ti Tey

Research output: Contribution to journalArticleResearchpeer-review

11 Citations (Scopus)

Abstract

HBcAg (hepatitis B core antigen) is a nanoplex bioproduct that has a great potential in the development of therapeutic drugs and vaccines. In the present study, a continuous-flow bead milling for the disruption of Escherichia coli was optimized and a direct recovery protocol to isolate the recombinant HBcAg from the unclarified E. coli disruptate was developed. The optimal condition for continuous-flow bead milling for the release of HBcAg from E. coli was achieved at a feed flow rate of 15 litres/h, biomass concentration of 10% [ww/v (wet weight/vol.)] and impeller tip speed of 14 m/s. The sucrose-density-gradient analysis showed that the particulate form of the HBcAg released by this optimal condition is still preserved. In the direct purification of HBcAg from the unclarified disruptate, the AE-EBAC (anion-exchange expanded-bed adsorption chromatography) technique was employed. A 54% adsorption and 50.7% recovery of HBcAg were achieved in this direct recovery process. The purity of HBcAg recovered was 49.8%, which corresponds to a purification factor of 2.0. ELISA showed that the HBcAg recovered is functionally active.

Original languageEnglish
Pages (from-to)49-59
Number of pages11
JournalBiotechnology and Applied Biochemistry
Volume50
Issue number1
DOIs
Publication statusPublished - May 2008
Externally publishedYes

Keywords

  • Anion-exchange expanded-bed adsorption chromatography (AE-EBAC)
  • Continuous-flow bead milling
  • Escherichia coli
  • Hepatitis B core antigen (HBcAg)
  • Hepatitis B virus (HBV)
  • Therapeutic vaccine

Cite this