Quantification of soluble epoxide hydrolase inhibitors in experimental and clinical samples using the nanobody-based ELISA
10.1016/j.jpha.2023.05.006
- Author:
Huiyi YANG
1
,
2
;
Meng QI
;
Qiyi HE
;
Hee-Sung HWANG
;
Jun YANG
;
Mark MCCOY
;
Christophe MORISSEAU
;
Suqing ZHAO
;
Bruce D.HAMMOCK
Author Information
1. Department of Pharmaceutical Engineering,School of Biomedical and Pharmaceutical Sciences,Guangdong University of Technology,Guangzhou,China
2. Department of Entomology and Nematology and UCD Comprehensive Cancer Center,University of California,Davis,CA 95616,USA
- Keywords:
Nanobody;
Immunoassay;
Soluble epoxide hydrolase inhibitors;
Metabolites;
Small molecules
- From:
Journal of Pharmaceutical Analysis
2023;13(9):1013-1023
- CountryChina
- Language:Chinese
-
Abstract:
To ensure proper dosage of a drug,analytical quantification of it in biofluid is necessary.Liquid chro-matography mass spectrometry(LC-MS)is the conventional method of choice as it permits accurate identification and quantification.However,it requires expensive instrumentation and is not appropriate for bedside use.Using soluble epoxide hydrolase(sEH)inhibitors(EC5026 and TPPU)as examples,we report development of a nanobody-based enzyme-linked immunosorbent assay(ELISA)for such small molecules and its use to accurately quantify the drug chemicals in human samples.Under optimized conditions,two nanobody-based ELISAs were successfully established for EC5026 and TPPU with low limits of detection of 0.085 ng/mL and 0.31 ng/mL,respectively,and two order of magnitude linear ranges with high precision and accuracy.The assay was designed to detect parent and two biologically active metabolites in the investigation of a new drug candidate EC5026.In addition,the ELISAs displayed excellent correlation with LC-MS analysis and evaluation of inhibitory potency.The results indicate that nanobody-based ELISA methods can efficiently analyze drug like compounds.These methods could be easily implemented by the bedside,in the field in remote areas or in veterinary practice.This work il-lustrates that nanobody based assays offer alternative and supplementary analytical tools to mass spectrometry for monitoring small molecule medicines during clinical development and therapy.At-tributes of nanobody based pharmaceutical assays are discussed.
