Hepatitis C virus expressing flag-tagged envelope protein 2 has unaltered infectivity and density, is specifically neutralized by flag antibodies and can be purified by affinity chromatography

Abstract Hepatitis C virus (HCV) purification by ultracentrifugation is difficult because of the low and heterogeneous density of native and cultured viruses. It was recently shown that inserting flag tag into envelope protein 2 (E2) of HCV permitted virus purification by affinity chromatography. Ho...

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Bibliographic Details
Published in:Virology (New York, N.Y.) N.Y.), 2011-01, Vol.409 (2), p.148-155
Main Authors: Prentoe, Jannick, Bukh, Jens
Format: Article
Language:English
Subjects:
Antibodies, Neutralizing - immunology
Antibodies, Viral - immunology
Cell Line
Chromatography, Affinity - methods
Envelope protein
Flag tag
Hepacivirus - genetics
Hepacivirus - isolation & purification
Hepacivirus - pathogenicity
Hepatitis C virus
Hepatocytes - virology
Humans
Immuno-staining
Infectious Disease
Microbial Viability
Mutagenesis, Insertional
Neutralization
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - immunology
Recombinant Fusion Proteins - metabolism
Staining and Labeling - methods
Vaccine
Viral Envelope Proteins - genetics
Viral Envelope Proteins - immunology
Viral Envelope Proteins - metabolism
Virulence
Virus purification
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Summary:Abstract Hepatitis C virus (HCV) purification by ultracentrifugation is difficult because of the low and heterogeneous density of native and cultured viruses. It was recently shown that inserting flag tag into envelope protein 2 (E2) of HCV permitted virus purification by affinity chromatography. However, flag-tagged viruses had drastically altered properties, and purification yield was low. In this study, we found that insertion of flag tag at the N-terminus of E2 in HCV recombinant J6/JFH1 did not affect viability in Huh7.5 cells, and that flag-tagged virus had physiochemical properties similar to the original virus. Flag-tagged virus was susceptible to flag-specific antibody neutralization, and infected cells could be immuno-stained by anti-flag antibodies. Using affinity chromatography with anti-flag resin we repeatedly obtained ~ 30% recovery of infectious particles. The full viability and unaltered physiochemical properties of flag-tagged HCV is an important improvement for utilizing these viruses for imaging, virion composition analysis and possibly vaccine development.
ISSN:0042-6822
1096-0341
DOI:10.1016/j.virol.2010.10.034