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US 8,877,434 B2Granted
Method and system of detecting dioxin-like compounds
OwnerHsinyu Leeinventor
Life of this patent
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Abstract
The invention relates to a method or a cell free system of detecting dioxin-like compounds in a test sample using a whole cell lysate derived from a cell transfected to express a fusion protein comprising an AHR fused to a reporter peptide. The method in combination with bioluminescence resonance energy transfer (BRET) technique is also provided so as to improve the sensitivity.
Claims9
A method of detecting dioxin or a dioxin-like compound in a test sample, wherein the dioxin-like compound is selected from the group consisting of polychlorinated dibenzodioxins, polychlorinated dibenzofurans, and polychlorinated biphenyls, comprising:
- providing a cell-free system comprising whole cell lysates that are non-viable cell suspensions obtained by culturing a cell line transfected to co-express an aryl hydrocarbon receptor (AHR) fused to a reporter peptide, and heat shock protein 90 (HSP90) to form a complex of AHR and HSP90 in the cell line, harvesting and lysing the whole cells, and removing the cell debris;
- contacting the test sample with the whole cell lysates for a period of time sufficient to induce a change of the detectable signal generated by the reporter peptide fused to AHR when the AHR degrades upon the presence of the dioxin or dioxin-like compound;
- and
- detecting the change of the detectable signal of the reporter peptide fused to AHR as indicative of the level of the degradation of AHR and comparing to a reference signal-generated by the cell free system with known concentrations of dioxin or a dioxin-like compound, thereby determining the presence or amount of the dioxin or dioxin-like compound in the test sample.
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Dependencies are inferred from the claim text.