N-terminally His-tagged hepatitis B core antigens: construction, expression, purification and antigenicity

J Virol Methods. 2009 Sep;160(1-2):125-31. doi: 10.1016/j.jviromet.2009.04.038. Epub 2009 May 9.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, Affinity
  • Escherichia coli / genetics
  • Gene Expression
  • Hepatitis B Core Antigens / genetics
  • Hepatitis B Core Antigens / immunology*
  • Hepatitis B Core Antigens / isolation & purification*
  • Microscopy, Electron, Transmission
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / isolation & purification
  • Virosomes / ultrastructure

Substances

  • Hepatitis B Core Antigens
  • Recombinant Fusion Proteins
  • Virosomes