title: Heavy chain single-domain antibodies to detect native human soluble epoxide hydrolase.
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.
authors.affiliation: Department of Entomology and Nematology, University of California, Davis, Davis, CA, 95616, USA.|UCD Comprehensive Cancer Center, University of California, Davis, Davis, CA, 95616, USA.
authors.forename: Yongliang
authors.initials: Y
authors.lastname: Cui
authors.affiliation: Department of Entomology and Nematology, University of California, Davis, Davis, CA, 95616, USA.|College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, Zhejiang, 310058, China.|UCD Comprehensive Cancer Center, University of California, Davis, Davis, CA, 95616, USA.
authors.forename: Dongyang
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Entomology and Nematology, University of California, Davis, Davis, CA, 95616, USA.|UCD Comprehensive Cancer Center, University of California, Davis, Davis, CA, 95616, USA.
authors.forename: Christophe
authors.initials: C
authors.lastname: Morisseau
authors.affiliation: Department of Entomology and Nematology, University of California, Davis, Davis, CA, 95616, USA.|UCD Comprehensive Cancer Center, University of California, Davis, Davis, CA, 95616, USA.
authors.forename: Jie-Xian
authors.initials: JX
authors.lastname: Dong
authors.affiliation: Department of Entomology and Nematology, University of California, Davis, Davis, CA, 95616, USA.|UCD Comprehensive Cancer Center, University of California, Davis, Davis, CA, 95616, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Entomology and Nematology, University of California, Davis, Davis, CA, 95616, USA.|UCD Comprehensive Cancer Center, University of California, Davis, Davis, CA, 95616, USA.
authors.forename: Debin
authors.initials: D
authors.lastname: Wan
authors.affiliation: Cátedra de Inmunología, Facultad de Química, Instituto de Higiene, UDELAR Av. A. Navarro 3051, Piso 2, Montevideo, 11600, Uruguay.
authors.forename: Martín A
authors.initials: MA
authors.lastname: Rossotti
authors.affiliation: Department of Entomology and Nematology, University of California, Davis, Davis, CA, 95616, USA.|UCD Comprehensive Cancer Center, University of California, Davis, Davis, CA, 95616, USA.
authors.forename: Shirley J
authors.initials: SJ
authors.lastname: Gee
authors.affiliation: Cátedra de Inmunología, Facultad de Química, Instituto de Higiene, UDELAR Av. A. Navarro 3051, Piso 2, Montevideo, 11600, Uruguay.
authors.forename: Gualberto G
authors.initials: GG
authors.lastname: González-Sapienza
authors.affiliation: Department of Entomology and Nematology, University of California, Davis, Davis, CA, 95616, USA. bdhammock@ucdavis.edu.|UCD Comprehensive Cancer Center, University of California, Davis, Davis, CA, 95616, USA. bdhammock@ucdavis.edu.
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Hammock
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002161:Camelids, New World; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D004851:Epoxide Hydrolases / Q000032:analysis* / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D061905:Single-Domain Antibodies / Q000737:chemistry / Q000276:immunology*
chemical_list: D061905:Single-Domain Antibodies; D004851:Epoxide Hydrolases
keywords: ELISA; Magnetic beads; Soluble epoxide hydrolase; VHH; sEH