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Characterization of monoclonal antibodies against the nonstructural 5A protein of hepatitis C virus

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Abstract

The nonstructural 5A (NS5A) protein of hepatitis C virus (HCV) is a multifunctional protein that leads to pleiotropic responses, in part by regulating cell growth and cellular signaling pathways. Here, we produced monoclonal antibodies (mAbs) directed against the HCV NS5A protein. The N-terminal epitope was mapped to amino acids 60–80 of the NS5A protein, and the epitope in the middle region was mapped to amino acids 221–236. Because these epitopes overlap with binding regions of human vesicle-associated membrane-protein-associated protein (hVAP)-B and TNF-receptor-associated factor 2 (TRAF2), respectively, we investigated these mAbs for their potential capacity to inhibit viral and cellular interactions. We found that NS5A and hVAP-B interaction was abolished by mAb E5D3, and NS5A and TRAF2 interaction was inhibited by mAb C6D4. Since hVAP-B is necessary for HCV replication and TRAF2 is the major transducer in TNF signaling cascades, these data may provide further insights into the mechanisms underlying HCV replication and viral modulation of host signal transduction.

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Abbreviations

HCV:

Hepatitis C virus

NS5A:

Nonstructural 5A

mAb:

Monoclonal antibody

Sf9:

Spodoptera frugiperda

hVAP-B:

Human vesicle-associated membrane protein-associated protein-B

TRAF2:

TNF receptor-associated factor 2

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Acknowledgments

The authors thank Dr. Seng-Lai Tan (Eli Lilly and Company) for providing us with rabbit anti-phospho NS5A antibodies. This study was supported by a grant of the Korea Health 21 R&D project, Ministry of Health and Welfare, Republic of Korea (A060032), Biotechnology Development (2008-04100), Research Program for New Drug Target Discovery (M10601000044-06N0100-04410), and NRL (ROA-2007-000-20051-0) from the Ministry of Education, Science and Technology, Korea.

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Authors and Affiliations

  1. National Research Laboratory of Hepatitis C Virus, Ilsong Institute of Life Science, Hallym University, 1605-4 Gwanyang-dong, Dongan-gu, Anyang, 431-060, Korea

    Sang Min Kang, Hyun-Jung Jun, Yun-Sook Lim, Soo-Ho Choi & Soon B. Hwang

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  1. Sang Min Kang
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  2. Hyun-Jung Jun
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  3. Yun-Sook Lim
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  4. Soo-Ho Choi
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  5. Soon B. Hwang
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Correspondence to Soon B. Hwang.

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Kang, S.M., Jun, HJ., Lim, YS. et al. Characterization of monoclonal antibodies against the nonstructural 5A protein of hepatitis C virus. Arch Virol 154, 843–851 (2009). https://doi.org/10.1007/s00705-009-0386-9

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  • DOI: https://doi.org/10.1007/s00705-009-0386-9

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