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Heavy chain single-domain antibodies to detect native human soluble epoxide hydrolase

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Abstract

The soluble epoxide hydrolase (sEH) is a potential pharmacological target for treating hypertension, vascular inflammation, pain, cancer, and other diseases. However, there is not a simple, inexpensive, and reliable method to estimate levels of active sEH in tissues. Toward developing such an assay, a polyclonal variable domain of heavy chain antibody (VHH) sandwich immunoassay was developed. Ten VHHs, which are highly selective for native human sEH, were isolated from a phage-displayed library. The ten VHHs have no significant cross-reactivity with human microsomal epoxide hydrolase, rat and mouse sEH, and denatured human sEH. There is a high correlation between protein levels of the sEH determined by the enzyme-linked immunosorbent assay (ELISA) and the catalytic activity of the enzyme in S9 fractions of human tissues (liver, kidney, and lung). The VHH-based ELISA appears to be a new reliable method for monitoring the sEH and may be useful as a diagnostic tool for diseases influenced by sEH. This study also demonstrates the broad utility of VHH in biochemical and pharmacological research.

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Acknowledgments

This work was supported by the National Institute of Environmental Health Sciences (NIEHS) Superfund Research Program grant P42 ES04699, NIEHS R01 ES02710, FCE 6812 ANII (Agencia Nacional de Investigación e Innovación, Uruguay) and TW05718 Fogarty Center NHI. MR is a recipient of scholarships from ANII and CSIC, Uruguay.

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Authors and Affiliations

  1. Department of Entomology and Nematology, University of California, Davis, Davis, CA, 95616, USA

    Yongliang Cui, Dongyang Li, Christophe Morisseau, Jie-Xian Dong, Jun Yang, Debin Wan, Shirley J. Gee & Bruce D. Hammock

  2. College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, Zhejiang, 310058, China

    Dongyang Li

  3. Cátedra de Inmunología, Facultad de Química, Instituto de Higiene, UDELAR Av. A. Navarro 3051, Piso 2, Montevideo, 11600, Uruguay

    Martín A. Rossotti & Gualberto G. González-Sapienza

  4. UCD Comprehensive Cancer Center, University of California, Davis, Davis, CA, 95616, USA

    Yongliang Cui, Dongyang Li, Christophe Morisseau, Jie-Xian Dong, Jun Yang, Debin Wan, Shirley J. Gee & Bruce D. Hammock

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  1. Yongliang Cui
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  2. Dongyang Li
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Correspondence to Bruce D. Hammock.

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Cui, Y., Li, D., Morisseau, C. et al. Heavy chain single-domain antibodies to detect native human soluble epoxide hydrolase. Anal Bioanal Chem 407, 7275–7283 (2015). https://doi.org/10.1007/s00216-015-8889-6

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  • DOI: https://doi.org/10.1007/s00216-015-8889-6

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