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The Generation of a Nanobody-Based ELISA for Human Microsomal Epoxide Hydrolase.
He, Qiyi; McCoy, Mark R; Qi, Meng; Morisseau, Christophe; Yang, Huiyi; Xu, Chengpeng; Shey, Rachel; Goodman, Michael C; Zhao, Suqing; Hammock, Bruce D.
Affiliation
  • He Q; Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA.
  • McCoy MR; Department of Pharmaceutical Engineering, School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, China.
  • Qi M; Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA.
  • Morisseau C; Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA.
  • Yang H; Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA.
  • Xu C; Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA.
  • Shey R; Department of Pharmaceutical Engineering, School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, China.
  • Goodman MC; Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA.
  • Zhao S; Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA.
  • Hammock BD; Department of Entomology and Nematology and University of California Davis Comprehensive Cancer Center, University of California Davis, Davis, CA 95616, USA.
Int J Mol Sci ; 24(19)2023 Sep 28.
Article in En | MEDLINE | ID: mdl-37834144
ABSTRACT
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.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Epoxide Hydrolases / Single-Domain Antibodies Limits: Humans Language: En Journal: Int J Mol Sci Year: 2023 Document type: Article Affiliation country: Estados Unidos
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Epoxide Hydrolases / Single-Domain Antibodies Limits: Humans Language: En Journal: Int J Mol Sci Year: 2023 Document type: Article Affiliation country: Estados Unidos