N-terminally His-tagged hepatitis B core antigens

Construction, expression, purification and antigenicity

Wei Boon Yap, Beng Ti Tey, Michelle Y T Ng, Swee Tin Ong, Wen Siang Tan

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The core antigen of the hepatitis B virus (HBcAg) has been used widely as a diagnostic reagent for the identification of the viral infection. However, purification using the conventional sucrose density gradient ultracentrifugation is time consuming and costly. To overcome this, HBcAg particles displaying His-tag on their surface were constructed and produced in Escherichia coli. The recombinant His-tagged HBcAgs were purified using immobilized metal affinity chromatography. Transmission electron microscopy and enzyme-linked immunosorbent assay (ELISA) revealed that the displayed His-tag did not impair the formation of the core particles and the antigenicity of HBcAg.

Original languageEnglish
Pages (from-to)125-131
Number of pages7
JournalJournal of Virological Methods
Volume160
Issue number1-2
DOIs
Publication statusPublished - Sep 2009
Externally publishedYes

Fingerprint

Hepatitis B Core Antigens
Ultracentrifugation
Virus Diseases
Transmission Electron Microscopy
Affinity Chromatography
Hepatitis B virus
Sucrose
Metals
Enzyme-Linked Immunosorbent Assay
Escherichia coli
Antigens

Keywords

  • Antigenicity
  • Hepatitis B capsid
  • His-tag
  • Protein display
  • Purification

ASJC Scopus subject areas

  • Virology

Cite this

N-terminally His-tagged hepatitis B core antigens : Construction, expression, purification and antigenicity. / Yap, Wei Boon; Tey, Beng Ti; Ng, Michelle Y T; Ong, Swee Tin; Tan, Wen Siang.

In: Journal of Virological Methods, Vol. 160, No. 1-2, 09.2009, p. 125-131.

Research output: Contribution to journalArticle

Yap, Wei Boon ; Tey, Beng Ti ; Ng, Michelle Y T ; Ong, Swee Tin ; Tan, Wen Siang. / N-terminally His-tagged hepatitis B core antigens : Construction, expression, purification and antigenicity. In: Journal of Virological Methods. 2009 ; Vol. 160, No. 1-2. pp. 125-131.
@article{76b11916773246c69d249eeabda1519c,
title = "N-terminally His-tagged hepatitis B core antigens: Construction, expression, purification and antigenicity",
abstract = "The core antigen of the hepatitis B virus (HBcAg) has been used widely as a diagnostic reagent for the identification of the viral infection. However, purification using the conventional sucrose density gradient ultracentrifugation is time consuming and costly. To overcome this, HBcAg particles displaying His-tag on their surface were constructed and produced in Escherichia coli. The recombinant His-tagged HBcAgs were purified using immobilized metal affinity chromatography. Transmission electron microscopy and enzyme-linked immunosorbent assay (ELISA) revealed that the displayed His-tag did not impair the formation of the core particles and the antigenicity of HBcAg.",
keywords = "Antigenicity, Hepatitis B capsid, His-tag, Protein display, Purification",
author = "Yap, {Wei Boon} and Tey, {Beng Ti} and Ng, {Michelle Y T} and Ong, {Swee Tin} and Tan, {Wen Siang}",
year = "2009",
month = "9",
doi = "10.1016/j.jviromet.2009.04.038",
language = "English",
volume = "160",
pages = "125--131",
journal = "Journal of Virological Methods",
issn = "0166-0934",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - N-terminally His-tagged hepatitis B core antigens

T2 - Construction, expression, purification and antigenicity

AU - Yap, Wei Boon

AU - Tey, Beng Ti

AU - Ng, Michelle Y T

AU - Ong, Swee Tin

AU - Tan, Wen Siang

PY - 2009/9

Y1 - 2009/9

N2 - The core antigen of the hepatitis B virus (HBcAg) has been used widely as a diagnostic reagent for the identification of the viral infection. However, purification using the conventional sucrose density gradient ultracentrifugation is time consuming and costly. To overcome this, HBcAg particles displaying His-tag on their surface were constructed and produced in Escherichia coli. The recombinant His-tagged HBcAgs were purified using immobilized metal affinity chromatography. Transmission electron microscopy and enzyme-linked immunosorbent assay (ELISA) revealed that the displayed His-tag did not impair the formation of the core particles and the antigenicity of HBcAg.

AB - The core antigen of the hepatitis B virus (HBcAg) has been used widely as a diagnostic reagent for the identification of the viral infection. However, purification using the conventional sucrose density gradient ultracentrifugation is time consuming and costly. To overcome this, HBcAg particles displaying His-tag on their surface were constructed and produced in Escherichia coli. The recombinant His-tagged HBcAgs were purified using immobilized metal affinity chromatography. Transmission electron microscopy and enzyme-linked immunosorbent assay (ELISA) revealed that the displayed His-tag did not impair the formation of the core particles and the antigenicity of HBcAg.

KW - Antigenicity

KW - Hepatitis B capsid

KW - His-tag

KW - Protein display

KW - Purification

UR - http://www.scopus.com/inward/record.url?scp=67649119641&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67649119641&partnerID=8YFLogxK

U2 - 10.1016/j.jviromet.2009.04.038

DO - 10.1016/j.jviromet.2009.04.038

M3 - Article

VL - 160

SP - 125

EP - 131

JO - Journal of Virological Methods

JF - Journal of Virological Methods

SN - 0166-0934

IS - 1-2

ER -