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Hepatitis C Virus Protocols pp 295–304Cite as

Simultaneous Quantification of Hepatitis C Virus Envelope Glycoproteins E1 and E2 by Dual-Color Fluorescence Immunoblot Analysis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1911))

Abstract

The hepatitis C virus (HCV) envelope glycoproteins, E1 and E2, are crucial for HCV assembly and entry, and are promising vaccine antigens. However, they are challenging to study because of technical difficulties in protein production and in quality control for protein folding and glycosylation. To study E1 and E2 in different experimental systems, e.g. infected cells, virus culture, virus-like particles, and clinical samples, a standardized method to accurately quantify the glycoproteins will be essential for most research projects. Here we outline a sensitive assay based on dual-color fluorescence immunoblot and the Odyssey imaging system to detect and quantify HCV E1 and E2 glycoproteins either using a purified E1E2 complex, or an engineered protein standard containing E1 and E2 at equal molar ratio. The method is capable of simultaneously detecting and quantifying as little as 7 ng of E1 and 5 ng of E2 in HCV pseudoparticles, and will be useful to quantify E1 and E2 from a wide variety of samples.

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Acknowledgments

This work was supported by NIH grants AI079031, AI106005, AI123365, and AI123861.

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Authors and Affiliations

  1. Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, USA

    Fang Chen, Erick Giang, Radhika Gopal & Mansun Law

Authors
  1. Fang Chen
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  2. Erick Giang
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  3. Radhika Gopal
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  4. Mansun Law
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Corresponding author

Correspondence to Mansun Law .

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Editors and Affiliations

  1. Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, USA

    Mansun Law

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Chen, F., Giang, E., Gopal, R., Law, M. (2019). Simultaneous Quantification of Hepatitis C Virus Envelope Glycoproteins E1 and E2 by Dual-Color Fluorescence Immunoblot Analysis. In: Law, M. (eds) Hepatitis C Virus Protocols . Methods in Molecular Biology, vol 1911. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8976-8_20

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  • DOI: https://doi.org/10.1007/978-1-4939-8976-8_20

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8975-1

  • Online ISBN: 978-1-4939-8976-8

  • eBook Packages: Springer Protocols

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