title: The direct recovery of recombinant hepatitis B core antigen from disruptate derived from continuous-flow bead milling.
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.
authors.affiliation: Department of Chemical and Environmental Engineering, Faculty of Engineering, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia.
authors.forename: Chin Woi
authors.initials: CW
authors.lastname: Ho
authors.forename: Wen Siang
authors.initials: WS
authors.lastname: Tan
authors.forename: Suryani
authors.initials: S
authors.lastname: Kamaruddin
authors.forename: Tau Chuan
authors.initials: TC
authors.lastname: Ling
authors.forename: Beng Ti
authors.initials: BT
authors.lastname: Tey
mesh_terms: D000327:Adsorption; D018533:Biomass; D002852:Chromatography, Ion Exchange; D004926:Escherichia coli / Q000166:cytology / Q000378:metabolism; D006512:Hepatitis B Core Antigens / Q000302:isolation & purification*; D042541:Intracellular Space / Q000378:metabolism; D007700:Kinetics; D011994:Recombinant Proteins / Q000302:isolation & purification*
chemical_list: D006512:Hepatitis B Core Antigens; D011994:Recombinant Proteins