The Generation of a Nanobody-Based ELISA for Human Microsomal Epoxide Hydrolase

Qiyi He; Mark R. McCoy; Meng Qi; Christophe Morisseau; Huiyi Yang; Chengpeng Xu; Rachel Shey; Michael C. Goodman; Suqing Zhao; Bruce D. Hammock

doi:10.3390/ijms241914698

International Journal of Molecular Sciences (Sep 2023)

The Generation of a Nanobody-Based ELISA for Human Microsomal Epoxide Hydrolase

Qiyi He,
Mark R. McCoy,
Meng Qi,
Christophe Morisseau,
Huiyi Yang,
Chengpeng Xu,
Rachel Shey,
Michael C. Goodman,
Suqing Zhao,
Bruce D. Hammock

Affiliations

Qiyi He: Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA
Mark R. McCoy: Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA
Meng Qi: Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA
Christophe Morisseau: Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA
Huiyi Yang: Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA
Chengpeng Xu: Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA
Rachel Shey: Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA
Michael C. Goodman: Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA
Suqing Zhao: Department of Pharmaceutical Engineering, School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, China
Bruce D. Hammock: Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA

DOI: https://doi.org/10.3390/ijms241914698
Journal volume & issue: Vol. 24, no. 19
p. 14698

Abstract

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A microsomal epoxide hydrolase (mEH) metabolizes in vivo in both xenobiotic and endogenous epoxides associated with signaling function. Findings in patients suggest that mEH might be a biomarker for several diseases, including metastatic cancer and viral hepatitis. To easily quantify mEH, nanobodies specific to the human mEH were isolated from a phage library of llama VHHs. Four unique clones were obtained and used for developing ELISAs. Three formats of double antibody sandwich assays were investigated using different detection strategies. Using PolyHRP, the signal was strongly amplified, yielding a 22-fold lower LOD (12 pg mL−1) than the ‘conventional’. To further validate the performance of the immunoassays, human tissue samples were analyzed by nanobody-based ELISAs and compared to the enzyme activities (R2 > 0.95). The results demonstrate that these nanobodies are powerful tools for the quantification of human mEH and could eventually result in a bedside assay.

Published in International Journal of Molecular Sciences

ISSN: 1661-6596 (Print); 1422-0067 (Online)
Publisher: MDPI AG
Country of publisher: Switzerland
LCC subjects: Science: Biology (General); Science: Chemistry
Website: http://www.mdpi.com/journal/ijms

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