Preparation of hepatitis C virus structural and non-structural protein fragments and studies of their immunogenicity

Protein Expr Purif. 2006 Nov;50(1):43-8. doi: 10.1016/j.pep.2006.06.011. Epub 2006 Jun 27.

Abstract

Plasmids pQE-60 and pQE-30 containing 6 x His-tag sequence were used for expression of fragments of HCV structural and non-structural proteins in Escherichia coli (E. coli). The following fragments were used: core (1-98 aa), NS3 (202-482 aa), and tetramer of hypervariable region 1 (HVR1) of E2 protein. The constructed plasmids directed high levels of expression of HCV proteins in E. coli JM109. After purification by the metal-affinity chromatography on nickel-nitrilotriacetic acid (Ni-NTA) agarose, the His-tagged HCV proteins were used for immunization of BALB/c mice. All three proteins were able to induce high levels of specific antibodies and, in the case of the NS3 and HVR1 tetramer, also to mount vigorous cell-proliferating responses. High immunogenicity of the tested fragments of HCV proteins shows them as good candidates for inclusion into the future HCV vaccine preparations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigen-Antibody Reactions
  • Cell Proliferation
  • Chromatography, Affinity / methods
  • Cloning, Molecular
  • Dose-Response Relationship, Immunologic
  • Female
  • Hepatitis C Antibodies / immunology
  • Hepatitis C Antigens / genetics
  • Hepatitis C Antigens / immunology
  • Hepatitis C Antigens / isolation & purification*
  • Immunization
  • Mice
  • Mice, Inbred BALB C
  • Nickel / chemistry
  • Nitrilotriacetic Acid / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology
  • Peptide Fragments / isolation & purification*
  • Peptides / genetics
  • Peptides / immunology
  • Peptides / isolation & purification*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / isolation & purification
  • Sensitivity and Specificity
  • Spleen / cytology
  • Spleen / immunology
  • Viral Core Proteins / genetics
  • Viral Core Proteins / immunology
  • Viral Core Proteins / isolation & purification*
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / immunology
  • Viral Nonstructural Proteins / isolation & purification*
  • Viral Proteins / genetics
  • Viral Proteins / immunology
  • Viral Proteins / isolation & purification*

Substances

  • HVR1 protein, Hepatitis C virus
  • Hepatitis C Antibodies
  • Hepatitis C Antigens
  • NS3 protein, hepatitis C virus
  • Peptide Fragments
  • Peptides
  • Recombinant Proteins
  • Viral Core Proteins
  • Viral Nonstructural Proteins
  • Viral Proteins
  • core protein (1-98), hepatitis C virus
  • core protein p22, Hepatitis C virus
  • Nickel
  • Nitrilotriacetic Acid