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Highly sensitive reporter gene assay for dioxins in human plasma by using cycloheximide as an enhancer substance
Koichi Saito1, Haruyuki Matsunaga, Atsuko Ohmura
1Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Hoshi University, Shinagawa, Tokyo, Japan. k-saito@hoshi.ac.jp
Abstract:
A reporter gene assay (RGA) that uses a mouse liver recombinant Hepa1c1c7 containing the firefly luciferase gene was developed to screen for dioxins in human plasma. For a high-sensitivity method, the addition of cycloheximide to the culture medium brought about a fivefold increase in the sensitivity. The detection limit was 0.1 pg/microL/well. Aryl hydrocarbon receptor (AhR) binding affinity factors (AhR-BAF), calculated from the effect concentration 50 (EC(50)) value, showed approximately the same values as those in WHO-TEF (2006). A significant correlation between RGA and the conventional gas chromatography/mass spectrometry (GC/MS) method was obtained.
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