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Standardized Method for the Study of Antibody Neutralization of HCV Pseudoparticles (HCVpp)

  • Justin R. Bailey
  • Richard A. Urbanowicz
  • Jonathan K. Ball
  • Mansun Law
  • Steven K. H. Foung
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1911)

Abstract

Hepatitis C virus (HCV) pseudoparticles (HCVpp) are generated by cotransfection of HCV envelope (E1 and E2) genes along with a retroviral packaging/reporter construct into HEK293T cells. Enveloped particles bearing HCV E1E2 proteins on their surface are released through a retroviral budding process into the supernatant. Viral E1E2 glycoproteins facilitate a single round of receptor-mediated entry of HCVpp into hepatoma cells, which can be quantified by reporter gene expression. These HCVpp have been employed to study mechanisms of HCV entry into hepatoma cells, as well as HCV neutralization by immune sera or HCV-specific monoclonal antibodies.

Key words

Hepatitis C virus Flaviviridae HCV pseudoparticles Neutralizing antibodies Neutralizing breadth 

Notes

Acknowledgments

The authors would like to acknowledge Dr. Stuart Ray for useful discussions. This work was funded by NIH grants R01AI127469, R01AI079031, R01AI106005, U19AI123861, and U19AI088791, Medical Research Council (MRC) grant G0801169 and EU Seventh Framework Programme (FP7) grant “HepaMab” 305600.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Justin R. Bailey
    • 1
  • Richard A. Urbanowicz
    • 2
    • 5
  • Jonathan K. Ball
    • 2
  • Mansun Law
    • 3
  • Steven K. H. Foung
    • 4
  1. 1.Division of Infectious Diseases, Department of MedicineJohns Hopkins UniversityBaltimoreUSA
  2. 2.School of Life SciencesThe University of NottinghamNottinghamUK
  3. 3.Department of Immunology and MicrobiologyThe Scripps Research InstituteLa JollaUSA
  4. 4.Department of PathologyStanford University School of MedicineStanfordUSA
  5. 5.NIHR Biomedical Research Unit in Gastrointestinal and Liver Diseases, Nottingham University Hospitals NHS TrustUniversity of NottinghamNottinghamUK

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