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

被引:2
|
作者
He, Qiyi [1 ,2 ,3 ]
Mccoy, Mark R. [1 ,2 ]
Qi, Meng [1 ,2 ]
Morisseau, Christophe [1 ,2 ]
Yang, Huiyi [1 ,2 ,3 ]
Xu, Chengpeng [1 ,2 ]
Shey, Rachel [1 ,2 ]
Goodman, Michael C. [1 ,2 ]
Zhao, Suqing [2 ,3 ]
Hammock, Bruce D. [1 ,2 ]
机构
[1] Univ Calif Davis, Dept Entomol & Nematol, Davis, CA 95616 USA
[2] Univ Calif Davis, Comprehens Canc Ctr, Davis, CA 95616 USA
[3] Guangdong Univ Technol, Sch Biomed & Pharmaceut Sci, Dept Pharmaceut Engn, Guangzhou 510006, Peoples R China
关键词
nanobody; microsomal epoxide hydrolase; ELISA; biomarker; PRENEOPLASTIC ANTIGEN; EXPRESSION; ANTIBODIES; METABOLISM; CANCER; GENE;
D O I
10.3390/ijms241914698
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 (R-2 > 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.
引用
收藏
页数:12
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