@article{8b33ceb584e54dd9aa4cbbc2d724fe01,
title = "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. 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.",
keywords = "Antibodies, Neutralizing, Antibodies, Viral, Cell Line, Chromatography, Affinity, Hepacivirus, Hepatocytes, Humans, Microbial Viability, Mutagenesis, Insertional, Recombinant Fusion Proteins, Staining and Labeling, Viral Envelope Proteins, Virulence",
author = "Jannick Prentoe and Jens Bukh",
note = "Copyright {\textcopyright} 2010 Elsevier Inc. All rights reserved.",
year = "2011",
month = jan,
doi = "10.1016/j.virol.2010.10.034",
language = "English",
volume = "409",
pages = "148--55",
journal = "Virology",
issn = "1089-862X",
publisher = "Academic Press",
number = "2",
}