Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography

Wei Boon Yap; Beng Ti Tey; Noorjahan Banu Mohamed Alitheen; Wen Siang Tan

doi:10.1016/j.chroma.2010.03.012

Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography

Wei Boon Yap, Beng Ti Tey, Noorjahan Banu Mohamed Alitheen, Wen Siang Tan

Research output: Contribution to journal › Article › Research › peer-review

27 Citations (Scopus)

Abstract

Hepatitis B core antigen (HBcAg) is used as a diagnostic reagent for the detection of hepatitis B virus infection. In this study, immobilized metal affinity-expanded bed adsorption chromatography (IMA-EBAC) was employed to purify N-terminally His-tagged HBcAg from unclarified bacterial homogenate. Streamline Chelating was used as the adsorbent and the batch adsorption experiment showed that the optimal binding pH of His-tagged HBcAg was 8.0 with a binding capacity of 1.8mg per ml of adsorbent. The optimal elution condition for the elution of His-tagged HBcAg from the adsorbent was at pH 7 in the presence of 500mM imidazole and 1.5. M NaCl. The IMA-EBAC has successfully recovered 56% of His-tagged HBcAg from the unclarified E. coli homogenate with a purification factor of 3.64. Enzyme-linked immunosorbent assay (ELISA) showed that the antigenicity of the recovered His-tagged HBcAg was not affected throughout the IMA-EBAC purification process and electron microscopy revealed that the protein assembled into virus-like particles (VLP).

Original language	English
Pages (from-to)	3473-3480
Number of pages	8
Journal	Journal of Chromatography A
Volume	1217
Issue number	21
DOIs	https://doi.org/10.1016/j.chroma.2010.03.012
Publication status	Published - May 2010
Externally published	Yes

Keywords

His-tagged HBcAg
IMA-EBAC
Streamline Chelating
Unclarified bacterial homogenate
Virus-like particles

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.chroma.2010.03.012

Cite this

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title = "Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography",

abstract = "Hepatitis B core antigen (HBcAg) is used as a diagnostic reagent for the detection of hepatitis B virus infection. In this study, immobilized metal affinity-expanded bed adsorption chromatography (IMA-EBAC) was employed to purify N-terminally His-tagged HBcAg from unclarified bacterial homogenate. Streamline Chelating was used as the adsorbent and the batch adsorption experiment showed that the optimal binding pH of His-tagged HBcAg was 8.0 with a binding capacity of 1.8mg per ml of adsorbent. The optimal elution condition for the elution of His-tagged HBcAg from the adsorbent was at pH 7 in the presence of 500mM imidazole and 1.5. M NaCl. The IMA-EBAC has successfully recovered 56% of His-tagged HBcAg from the unclarified E. coli homogenate with a purification factor of 3.64. Enzyme-linked immunosorbent assay (ELISA) showed that the antigenicity of the recovered His-tagged HBcAg was not affected throughout the IMA-EBAC purification process and electron microscopy revealed that the protein assembled into virus-like particles (VLP).",

keywords = "His-tagged HBcAg, IMA-EBAC, Streamline Chelating, Unclarified bacterial homogenate, Virus-like particles",

author = "Yap, {Wei Boon} and Tey, {Beng Ti} and Alitheen, {Noorjahan Banu Mohamed} and Tan, {Wen Siang}",

note = "Funding Information: This study was supported by a FRGS grant (project no. 01-01-07-161FR) from the Ministry of Higher Education, Malaysia. W.B. Yap is supported by the Graduate Research Fellowship (GRF) from Universiti Putra Malaysia. Copyright: Copyright 2010 Elsevier B.V., All rights reserved.",

year = "2010",

month = may,

doi = "10.1016/j.chroma.2010.03.012",

language = "English",

volume = "1217",

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Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography. / Yap, Wei Boon; Tey, Beng Ti; Alitheen, Noorjahan Banu Mohamed et al.

In: Journal of Chromatography A, Vol. 1217, No. 21, 05.2010, p. 3473-3480.

Research output: Contribution to journal › Article › Research › peer-review

TY - JOUR

T1 - Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography

AU - Yap, Wei Boon

AU - Tey, Beng Ti

AU - Alitheen, Noorjahan Banu Mohamed

AU - Tan, Wen Siang

N1 - Funding Information: This study was supported by a FRGS grant (project no. 01-01-07-161FR) from the Ministry of Higher Education, Malaysia. W.B. Yap is supported by the Graduate Research Fellowship (GRF) from Universiti Putra Malaysia. Copyright: Copyright 2010 Elsevier B.V., All rights reserved.

PY - 2010/5

Y1 - 2010/5

N2 - Hepatitis B core antigen (HBcAg) is used as a diagnostic reagent for the detection of hepatitis B virus infection. In this study, immobilized metal affinity-expanded bed adsorption chromatography (IMA-EBAC) was employed to purify N-terminally His-tagged HBcAg from unclarified bacterial homogenate. Streamline Chelating was used as the adsorbent and the batch adsorption experiment showed that the optimal binding pH of His-tagged HBcAg was 8.0 with a binding capacity of 1.8mg per ml of adsorbent. The optimal elution condition for the elution of His-tagged HBcAg from the adsorbent was at pH 7 in the presence of 500mM imidazole and 1.5. M NaCl. The IMA-EBAC has successfully recovered 56% of His-tagged HBcAg from the unclarified E. coli homogenate with a purification factor of 3.64. Enzyme-linked immunosorbent assay (ELISA) showed that the antigenicity of the recovered His-tagged HBcAg was not affected throughout the IMA-EBAC purification process and electron microscopy revealed that the protein assembled into virus-like particles (VLP).

AB - Hepatitis B core antigen (HBcAg) is used as a diagnostic reagent for the detection of hepatitis B virus infection. In this study, immobilized metal affinity-expanded bed adsorption chromatography (IMA-EBAC) was employed to purify N-terminally His-tagged HBcAg from unclarified bacterial homogenate. Streamline Chelating was used as the adsorbent and the batch adsorption experiment showed that the optimal binding pH of His-tagged HBcAg was 8.0 with a binding capacity of 1.8mg per ml of adsorbent. The optimal elution condition for the elution of His-tagged HBcAg from the adsorbent was at pH 7 in the presence of 500mM imidazole and 1.5. M NaCl. The IMA-EBAC has successfully recovered 56% of His-tagged HBcAg from the unclarified E. coli homogenate with a purification factor of 3.64. Enzyme-linked immunosorbent assay (ELISA) showed that the antigenicity of the recovered His-tagged HBcAg was not affected throughout the IMA-EBAC purification process and electron microscopy revealed that the protein assembled into virus-like particles (VLP).

KW - His-tagged HBcAg

KW - IMA-EBAC

KW - Streamline Chelating

KW - Unclarified bacterial homogenate

KW - Virus-like particles

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M3 - Article

C2 - 20388569

AN - SCOPUS:77952553509

VL - 1217

SP - 3473

EP - 3480

JO - Journal of Chromatography A

JF - Journal of Chromatography A

SN - 0021-9673

IS - 21

ER -

Purification of His-tagged hepatitis B core antigen from unclarified bacterial homogenate using immobilized metal affinity-expanded bed adsorption chromatography

Abstract

Keywords

UN SDGs

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