Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography

Hepatitis B core antigen with the mutation of Y132A (HBcAg-Y132A) was successfully expressed in Escherichia coli. The mutant HBcAg-Y132A forms dimers and is unable to self-assemble into virus-like particles (VLPs). Hence, it is a potential antigen used in the antibody-responsive biosensor for the de...

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Main Authors:	Lim, Swee Lu, Ng, Hon Wei, Akwiditya, Made Angga, Ooi, Chien Wei, Chan, Eng-Seng, Ho, Kok Lian, Tan, Wen Siang, Chua, Gek Kee, Tey, Beng Ti
Format:	Article
Language:	English
Published:	Elsevier Ltd. 2018
Subjects:	TP Chemical technology
Online Access:	http://umpir.ump.edu.my/id/eprint/22999/ http://umpir.ump.edu.my/id/eprint/22999/ http://umpir.ump.edu.my/id/eprint/22999/ http://umpir.ump.edu.my/id/eprint/22999/1/Single-step%20purification%20of%20recombinant%20hepatitis%20B%20core%20antigen%20Y132A%20dimer.pdf

id	ump-22999
recordtype	eprints
spelling	ump-229992019-03-05T06:33:02Z http://umpir.ump.edu.my/id/eprint/22999/ Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography Lim, Swee Lu Ng, Hon Wei Akwiditya, Made Angga Ooi, Chien Wei Chan, Eng-Seng Ho, Kok Lian Tan, Wen Siang Chua, Gek Kee Tey, Beng Ti TP Chemical technology Hepatitis B core antigen with the mutation of Y132A (HBcAg-Y132A) was successfully expressed in Escherichia coli. The mutant HBcAg-Y132A forms dimers and is unable to self-assemble into virus-like particles (VLPs). Hence, it is a potential antigen used in the antibody-responsive biosensor for the detection of anti-HBcAg in patients infected with hepatitis B virus. The aim of this study was to establish a direct purification strategy to recover HBcAg-Y132A dimer from the E. coli feedstock using SepFast™ Supor DEAE pre-packed column. The performance of this anion exchange chromatography was optimized in terms of the buffer composition (for adsorption and elution steps) and the mode of elution (i.e., step or gradient). The highest adsorption of HBcAg-Y132A dimer in the DEAE column was achieved with the buffer composed of 50 mM Tris-HCl (pH 8.4). The step elution using 50 mM Tris-HCl elution buffer (pH 8.4) supplemented with 1 M NaCl resulted in 1.2-fold increase in the purity of HBcAg, as compared to the gradient elution mode. In addition, it was found that the optimized 3-step elution is not directly applicable to elute the self-assembled HBcAg VLPs, as only 24.7% of the particles were recovered due to the limitation of size effect. Elsevier Ltd. 2018 Article PeerReviewed pdf en http://umpir.ump.edu.my/id/eprint/22999/1/Single-step%20purification%20of%20recombinant%20hepatitis%20B%20core%20antigen%20Y132A%20dimer.pdf Lim, Swee Lu and Ng, Hon Wei and Akwiditya, Made Angga and Ooi, Chien Wei and Chan, Eng-Seng and Ho, Kok Lian and Tan, Wen Siang and Chua, Gek Kee and Tey, Beng Ti (2018) Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography. Process Biochemistry, 69. pp. 208-215. ISSN 1359-5113 https://doi.org/10.1016/j.procbio.2018.03.003 DOI: 10.1016/j.procbio.2018.03.003
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Malaysia Pahang
building	UMP Institutional Repository
collection	Online Access
language	English
topic	TP Chemical technology
spellingShingle	TP Chemical technology Lim, Swee Lu Ng, Hon Wei Akwiditya, Made Angga Ooi, Chien Wei Chan, Eng-Seng Ho, Kok Lian Tan, Wen Siang Chua, Gek Kee Tey, Beng Ti Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography
description	Hepatitis B core antigen with the mutation of Y132A (HBcAg-Y132A) was successfully expressed in Escherichia coli. The mutant HBcAg-Y132A forms dimers and is unable to self-assemble into virus-like particles (VLPs). Hence, it is a potential antigen used in the antibody-responsive biosensor for the detection of anti-HBcAg in patients infected with hepatitis B virus. The aim of this study was to establish a direct purification strategy to recover HBcAg-Y132A dimer from the E. coli feedstock using SepFast™ Supor DEAE pre-packed column. The performance of this anion exchange chromatography was optimized in terms of the buffer composition (for adsorption and elution steps) and the mode of elution (i.e., step or gradient). The highest adsorption of HBcAg-Y132A dimer in the DEAE column was achieved with the buffer composed of 50 mM Tris-HCl (pH 8.4). The step elution using 50 mM Tris-HCl elution buffer (pH 8.4) supplemented with 1 M NaCl resulted in 1.2-fold increase in the purity of HBcAg, as compared to the gradient elution mode. In addition, it was found that the optimized 3-step elution is not directly applicable to elute the self-assembled HBcAg VLPs, as only 24.7% of the particles were recovered due to the limitation of size effect.
format	Article
author	Lim, Swee Lu Ng, Hon Wei Akwiditya, Made Angga Ooi, Chien Wei Chan, Eng-Seng Ho, Kok Lian Tan, Wen Siang Chua, Gek Kee Tey, Beng Ti
author_facet	Lim, Swee Lu Ng, Hon Wei Akwiditya, Made Angga Ooi, Chien Wei Chan, Eng-Seng Ho, Kok Lian Tan, Wen Siang Chua, Gek Kee Tey, Beng Ti
author_sort	Lim, Swee Lu
title	Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography
title_short	Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography
title_full	Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography
title_fullStr	Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography
title_full_unstemmed	Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography
title_sort	single-step purification of recombinant hepatitis b core antigen y132a dimer from clarified escherichia coli feedstock using a packed bed anion exchange chromatography
publisher	Elsevier Ltd.
publishDate	2018
url	http://umpir.ump.edu.my/id/eprint/22999/ http://umpir.ump.edu.my/id/eprint/22999/ http://umpir.ump.edu.my/id/eprint/22999/ http://umpir.ump.edu.my/id/eprint/22999/1/Single-step%20purification%20of%20recombinant%20hepatitis%20B%20core%20antigen%20Y132A%20dimer.pdf
first_indexed	2023-09-18T22:34:16Z
last_indexed	2023-09-18T22:34:16Z
_version_	1777416490607181824

Single-step purification of recombinant hepatitis B core antigen Y132A dimer from clarified Escherichia coli feedstock using a packed bed anion exchange chromatography

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