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

• Main Authors: Ho, Chin Woi, Tan, Wen Siang, Kamarudin, Suryani, Ling, Tau Chuan, Tey, Beng Ti
• Format: Article
• Language: English
• Published: Portland Press 2008
• Online Access: http://psasir.upm.edu.my/id/eprint/17060/1/The%20direct%20recovery%20of%20recombinant%20hepatitis%20B%20core%20antigen%20from%20disruptate%20derived%20from%20continuous.pdf; http://psasir.upm.edu.my/id/eprint/17060/
• Institution: Universiti Putra Malaysia

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.