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

doi:10.1007/s00216-015-8889-6

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

Anal Bioanal Chem. 2015 Sep;407(24):7275-83. doi: 10.1007/s00216-015-8889-6. Epub 2015 Jul 31.

Authors

Yongliang Cui^{1

2}, Dongyang Li^{1

3

2}, Christophe Morisseau^{1

2}, Jie-Xian Dong^{1

2}, Jun Yang^{1

2}, Debin Wan^{1

2}, Martín A Rossotti⁴, Shirley J Gee^{1

2}, Gualberto G González-Sapienza⁴, Bruce D Hammock^{5

6}

Affiliations

¹ Department of Entomology and Nematology, University of California, Davis, Davis, CA, 95616, USA.
² UCD Comprehensive Cancer Center, University of California, Davis, Davis, CA, 95616, USA.
³ College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, Zhejiang, 310058, China.
⁴ Cátedra de Inmunología, Facultad de Química, Instituto de Higiene, UDELAR Av. A. Navarro 3051, Piso 2, Montevideo, 11600, Uruguay.
⁵ Department of Entomology and Nematology, University of California, Davis, Davis, CA, 95616, USA. bdhammock@ucdavis.edu.
⁶ UCD Comprehensive Cancer Center, University of California, Davis, Davis, CA, 95616, USA. bdhammock@ucdavis.edu.

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.

Keywords: ELISA; Magnetic beads; Soluble epoxide hydrolase; VHH; sEH.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Camelids, New World
Cross Reactions
Enzyme-Linked Immunosorbent Assay
Epoxide Hydrolases / analysis*
Epoxide Hydrolases / immunology
Humans
Molecular Sequence Data
Sequence Homology, Amino Acid
Single-Domain Antibodies / chemistry
Single-Domain Antibodies / immunology*

Substances

Single-Domain Antibodies
Epoxide Hydrolases

Abstract

Publication types

MeSH terms

Substances

Grants and funding