A preparative purification process for recombinant Hepatitis B core antigen using online capture by expanded bed adsorption followed by size-exclusion chromatography

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...

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Main Authors: Ho, Chin Woi, Tan, Wen Siang, Chong, Fui Chin, Ling, Tau Chuan, Tey, Beng Ti
Format: Article
Language:English
Published: Korean Society for Microbiology and Biotechnology 2009
Online Access:http://psasir.upm.edu.my/id/eprint/12790/
http://psasir.upm.edu.my/id/eprint/12790/1/A%20preparative%20purification%20process%20for%20recombinant%20Hepatitis%20B%20core%20antigen%20using%20online%20capture%20by%20expanded%20bed%20adsorption%20followed%20by%20size.pdf
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author Ho, Chin Woi
Tan, Wen Siang
Chong, Fui Chin
Ling, Tau Chuan
Tey, Beng Ti
author_facet Ho, Chin Woi
Tan, Wen Siang
Chong, Fui Chin
Ling, Tau Chuan
Tey, Beng Ti
author_sort Ho, Chin Woi
building UPM Institutional Repository
collection Online Access
description 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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spelling upm-127902015-09-17T00:10:47Z http://psasir.upm.edu.my/id/eprint/12790/ 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 Tan, Wen Siang Chong, Fui Chin Ling, Tau Chuan Tey, Beng Ti 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. Korean Society for Microbiology and Biotechnology 2009 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/12790/1/A%20preparative%20purification%20process%20for%20recombinant%20Hepatitis%20B%20core%20antigen%20using%20online%20capture%20by%20expanded%20bed%20adsorption%20followed%20by%20size.pdf Ho, Chin Woi and Tan, Wen Siang and Chong, Fui Chin and Ling, Tau Chuan and Tey, Beng Ti (2009) A preparative purification process for recombinant Hepatitis B core antigen using online capture by expanded bed adsorption followed by size-exclusion chromatography. Journal of Microbiology and Biotechnology, 19 (4). pp. 416-423. ISSN 1017-7825; ESSN: 1738-8872 http://www.jmb.or.kr/journal/viewJournal.html?year=2009&vol=19&num=4&page=416 10.4014/jmb.0804.254
spellingShingle Ho, Chin Woi
Tan, Wen Siang
Chong, Fui Chin
Ling, Tau Chuan
Tey, Beng Ti
A preparative purification process for recombinant Hepatitis B core antigen using online capture by expanded bed adsorption followed by size-exclusion chromatography
title A preparative purification process for recombinant Hepatitis B core antigen using online capture by expanded bed adsorption followed by size-exclusion chromatography
title_full A preparative purification process for recombinant Hepatitis B core antigen using online capture by expanded bed adsorption followed by size-exclusion chromatography
title_fullStr A preparative purification process for recombinant Hepatitis B core antigen using online capture by expanded bed adsorption followed by size-exclusion chromatography
title_full_unstemmed A preparative purification process for recombinant Hepatitis B core antigen using online capture by expanded bed adsorption followed by size-exclusion chromatography
title_short A preparative purification process for recombinant Hepatitis B core antigen using online capture by expanded bed adsorption followed by size-exclusion chromatography
title_sort preparative purification process for recombinant hepatitis b core antigen using online capture by expanded bed adsorption followed by size-exclusion chromatography
url http://psasir.upm.edu.my/id/eprint/12790/
http://psasir.upm.edu.my/id/eprint/12790/
http://psasir.upm.edu.my/id/eprint/12790/
http://psasir.upm.edu.my/id/eprint/12790/1/A%20preparative%20purification%20process%20for%20recombinant%20Hepatitis%20B%20core%20antigen%20using%20online%20capture%20by%20expanded%20bed%20adsorption%20followed%20by%20size.pdf