期刊
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
卷 158, 期 -, 页码 431-437出版社
ELSEVIER
DOI: 10.1016/j.jpba.2018.06.023
关键词
Glycocholic acid; Fluorescence polarization immunoassay; High-throughput screening; Polyclonal antibody; Urine
资金
- National Natural Science Foundation of China [41071176, 41271340]
- Guangdong Natural Science Foundation [S2013030013338]
- National Research Technical University MISiS [K3-2017-073]
- Third Affiliated Hospital of Sun Yat-sen University
In this paper, a simple, rapid and high-throughput fluorescence polarization immunoassay (FPIA) based on polyclonal antibodies (PAb) is described for the determination of glycocholic acid (GCA) in human urine. Three fluorescein-labeled GCA (tracers) with different structures and spacer bridges were synthesized and purified by thin-layer chromatography (TLC). The structure effect of tracers on the assay was investigated and the sensitivity of best tracer in the optimized FPIA demonstrated an IC50 value of 306 ng/mL. The working range of FPIA was 36 similar to 2 600 ng/mL and the limit of detection (LOD) was 9 ng/mL. The developed FPIA was time-saving that could be completed within 10 min. Human urine samples spiked with GCA were analyzed by this method, followed by confirmation with commercial enzyme immunoassay analysis (EIA). Excellent recoveries and correlation between these two methods were observed (R-2 = 0.996), suggesting the developed FPIA could be applied to screening of GCA in human urine samples without complicated cleanup. (C) 2018 Published by Elsevier B.V.
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