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

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

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. / Prentø, Jannick Cornelius; Bukh, Jens.

In: Virology, Vol. 409, No. 2, 2011, p. 148-55.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Prentø, JC & Bukh, J 2011, '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', Virology, vol. 409, no. 2, pp. 148-55. https://doi.org/10.1016/j.virol.2010.10.034

APA

Prentø, J. C., & Bukh, J. (2011). 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. Virology, 409(2), 148-55. https://doi.org/10.1016/j.virol.2010.10.034

Vancouver

Prentø JC, Bukh J. 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. Virology. 2011;409(2):148-55. https://doi.org/10.1016/j.virol.2010.10.034

Author

Prentø, Jannick Cornelius ; Bukh, Jens. / 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. In: Virology. 2011 ; Vol. 409, No. 2. pp. 148-55.

Bibtex

@article{8ab80dc14ba34ebf9524ef148bd7c7bc,
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.",
author = "Prent{\o}, {Jannick Cornelius} and Jens Bukh",
note = "Copyright {\textcopyright} 2010 Elsevier Inc. All rights reserved.",
year = "2011",
doi = "10.1016/j.virol.2010.10.034",
language = "English",
volume = "409",
pages = "148--55",
journal = "Virology",
issn = "0042-6822",
publisher = "Academic Press",
number = "2",

}

RIS

TY - JOUR

T1 - 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

AU - Prentø, Jannick Cornelius

AU - Bukh, Jens

N1 - Copyright © 2010 Elsevier Inc. All rights reserved.

PY - 2011

Y1 - 2011

N2 - 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.

AB - 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.

U2 - 10.1016/j.virol.2010.10.034

DO - 10.1016/j.virol.2010.10.034

M3 - Journal article

C2 - 21095619

VL - 409

SP - 148

EP - 155

JO - Virology

JF - Virology

SN - 0042-6822

IS - 2

ER -

ID: 34169492