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

Yongliang Cui, Dongyang Li, Christophe Morisseau, Jie Xian Dong, Jun Yang, Debin Wan, Martín A. Rossotti, Shirley J. Gee, Gualberto G. González-Sapienza, Bruce D. Hammock

Research output: Contribution to journalArticle

3 Scopus citations

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.

Original languageEnglish (US)
JournalAnalytical and Bioanalytical Chemistry
Volume407
Issue number24
DOIs
StatePublished - Jan 1 2017

Keywords

  • ELISA
  • Magnetic beads
  • sEH
  • Soluble epoxide hydrolase
  • VHH

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

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    Cui, Y., Li, D., Morisseau, C., Dong, J. X., Yang, J., Wan, D., Rossotti, M. A., Gee, S. J., González-Sapienza, G. G., & Hammock, B. D. (2017). Heavy chain single-domain antibodies to detect native human soluble epoxide hydrolase. Analytical and Bioanalytical Chemistry, 407(24). https://doi.org/10.1007/s00216-015-8889-6