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

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 recombina...

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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/
http://psasir.upm.edu.my/id/eprint/17060/1/The%20direct%20recovery%20of%20recombinant%20hepatitis%20B%20core%20antigen%20from%20disruptate%20derived%20from%20continuous.pdf
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author Ho, Chin Woi
Tan, Wen Siang
Kamarudin, Suryani
Ling, Tau Chuan
Tey, Beng Ti
author_facet Ho, Chin Woi
Tan, Wen Siang
Kamarudin, Suryani
Ling, Tau Chuan
Tey, Beng Ti
author_sort Ho, Chin Woi
building UPM Institutional Repository
collection Online Access
description 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.
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spelling upm-170602016-11-30T09:36:01Z http://psasir.upm.edu.my/id/eprint/17060/ The direct recovery of recombinant hepatitis B core antigen from disruptate derived from continuous-flow bead milling Ho, Chin Woi Tan, Wen Siang Kamarudin, Suryani Ling, Tau Chuan Tey, Beng Ti 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. Portland Press 2008 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/17060/1/The%20direct%20recovery%20of%20recombinant%20hepatitis%20B%20core%20antigen%20from%20disruptate%20derived%20from%20continuous.pdf Ho, Chin Woi and Tan, Wen Siang and Kamarudin, Suryani and Ling, Tau Chuan and Tey, Beng Ti (2008) The direct recovery of recombinant hepatitis B core antigen from disruptate derived from continuous-flow bead milling. Biotechnology and Applied Biochemistry, 50 (1). pp. 49-59. ISSN 0885-4513; ESSN: 1470-8744 10.1042/BA20070088
spellingShingle Ho, Chin Woi
Tan, Wen Siang
Kamarudin, Suryani
Ling, Tau Chuan
Tey, Beng Ti
The direct recovery of recombinant hepatitis B core antigen from disruptate derived from continuous-flow bead milling
title The direct recovery of recombinant hepatitis B core antigen from disruptate derived from continuous-flow bead milling
title_full The direct recovery of recombinant hepatitis B core antigen from disruptate derived from continuous-flow bead milling
title_fullStr The direct recovery of recombinant hepatitis B core antigen from disruptate derived from continuous-flow bead milling
title_full_unstemmed The direct recovery of recombinant hepatitis B core antigen from disruptate derived from continuous-flow bead milling
title_short The direct recovery of recombinant hepatitis B core antigen from disruptate derived from continuous-flow bead milling
title_sort direct recovery of recombinant hepatitis b core antigen from disruptate derived from continuous-flow bead milling
url http://psasir.upm.edu.my/id/eprint/17060/
http://psasir.upm.edu.my/id/eprint/17060/
http://psasir.upm.edu.my/id/eprint/17060/1/The%20direct%20recovery%20of%20recombinant%20hepatitis%20B%20core%20antigen%20from%20disruptate%20derived%20from%20continuous.pdf