6533b824fe1ef96bd1280125

RESEARCH PRODUCT

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

Melanie FiedlerMichael RoggendorfIvars PetrovskisMarija MihailovaSergei ViazovIrina SominskayaMechthild BoosPaul Pumpens

subject

Nitrilotriacetic AcidHepatitis C virusDose-Response Relationship ImmunologicViral Nonstructural ProteinsBiologymedicine.disease_causeSensitivity and SpecificityChromatography AffinityAntigen-Antibody ReactionsMiceViral Proteinschemistry.chemical_compoundPlasmidTetramerNickelmedicineAnimalsCloning MolecularEscherichia coliCell ProliferationMice Inbred BALB CNS3Viral Core ProteinsImmunogenicityvirus diseasesHepatitis C AntibodiesVirologyMolecular biologyPeptide FragmentsRecombinant Proteinsdigestive system diseasesHypervariable regionchemistryAgaroseFemaleImmunizationHepatitis C AntigensPeptidesSpleenBiotechnology

description

Abstract Plasmids pQE-60 and pQE-30 containing 6× 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.

yearjournalcountryeditionlanguage
2006-03-28Protein Expression and Purification
https://doi.org/10.1016/j.pep.2006.06.011