Enhancing the response of CALUX and CAFLUX cell bioassays for quantitative detection of dioxin-like compounds

Zhao, Bin; Baston, David S.; Khan, Elaine M.; Sorrentino, Claudio; Denison, Michael S.

doi:10.1007/s11426-010-0142-8

Cited by 29 publications

(15 citation statements)

References 17 publications

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“…22, 34–38 One application of our newly developed G3 CALUX cell lines is to examine species-specificity of chemicals (as pure compounds or present in known or unknown mixtures). While species-specific CALUX AhR bioassays are available, the majority of the validated (and commonly used) cell lines contain different luciferase reporter plasmids, complicating clear interpretation of results from common samples analyzed between cell lines.…”

Section: Resultsmentioning

confidence: 99%

Development of Species-Specific Ah Receptor-Responsive Third Generation CALUX Cell Lines with Enhanced Responsiveness and Improved Detection Limits

Brennan

Tsutsumi

et al. 2015

Environ. Sci. Technol.

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The Ah receptor (AhR)-responsive CALUX (chemically-activated luciferase expression) cell bioassay is commonly used for rapid screening of samples for the presence of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, dioxin), dioxin-like compounds, and AhR agonists/antagonists. By increasing the number of AhR DNA recognition sites (dioxin responsive elements), we previously generated a novel third generation (G3) recombinant AhR-responsive mouse CALUX cell line (H1L7.5c3) with significantly enhanced sensitivity and response to DLCs compared to existing AhR-CALUX cell bioassays. However, the elevated background luciferase activity of these cells and the absence of comparable G3 cell lines derived from other species have limited their utility for screening purposes. Here, we describe the development and characterization of species-specific G3 recombinant AhR-responsive CALUX cell lines (rat, human, and guinea pig) that exhibit significantly improved sensitivity and dramatically increased TCDD induction response. The low background luciferase activity, low minimal detection limit (0.1 pM TCDD) and enhanced induction response of the rat G3 cell line (H4L7.5c2) over the H1L7.5c3 mouse G3 cells, identifies them as a more optimal cell line for screening purposes. The utility of the new G3 CALUX cell lines were demonstrated by screening sediment extracts and a small chemical compound library for the presence of AhR agonists. The increased sensitivity and response of these new G3 CALUX cell lines will facilitate species-specific analysis of DLCs and AhR agonists in samples with low levels of contamination and/or in small sample volumes.

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Section: Resultsmentioning

confidence: 99%

Development of Species-Specific Ah Receptor-Responsive Third Generation CALUX Cell Lines with Enhanced Responsiveness and Improved Detection Limits

Brennan

Tsutsumi

et al. 2015

Environ. Sci. Technol.

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“…This yeast glucocorticoid bioassay was tested by exposures to cortisol and other related compounds and showed sensitivity to all chemicals, with the following order in relative potencies: budesonide > corticosterone > dexamethasone > cortisol = betamethasone > prednisolone > aldosterone. The obtained data were critically compared to a more sensitive GR-CALUX bioassay based on a human bone cell [9,49].…”

Section: Bacterial and Yeast Biosensors For Evaluating Potential Impamentioning

confidence: 99%

Environmental Application of Reporter-Genes Based Biosensors for Chemical Contamination Screening

Matejczyk¹,

Rosochacki²

2014

Archives of Environmental Protection

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Abstract:The paper presents results of research concerning possibilities of applications of reporter-genes based microorganisms, including the selective presentation of defects and advantages of different new scientifi c achievements of methodical solutions in genetic system constructions of biosensing elements for environmental research. The most robust and popular genetic fusion and new trends in reporter genes technology -such as LacZ (β-galactosidase), xylE (catechol 2,3-dioxygenase), gfp (green fl uorescent proteins) and its mutated forms, lux (prokaryotic luciferase), luc (eukaryotic luciferase), phoA (alkaline phosphatase), gusA and gurA (β-glucuronidase), antibiotics and heavy metals resistance are described. Reporter-genes based biosensors with use of genetically modifi ed bacteria and yeast successfully work for genotoxicity, bioavailability and oxidative stress assessment for detection and monitoring of toxic compounds in drinking water and different environmental samples, surface water, soil, sediments.

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“…[109]. These authors improved the sensitivity of the CALUX bioassay by monitoring the modulators of the cell signaling pathways and modification of cell culture conditions.…”

Section: Assays For Detecting Dioxins and Dl-pcbs In The Food Chainmentioning

confidence: 99%

The Need and Potential of Biosensors to Detect Dioxins and Dioxin-Like Polychlorinated Biphenyls along the Milk, Eggs and Meat Food Chain

Chobtang

Boer

Hoogenboom

et al. 2011

Sensors

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Dioxins and dioxin-like polychlorinated biphenyls (DL-PCBs) are hazardous toxic, ubiquitous and persistent chemical compounds, which can enter the food chain and accumulate up to higher trophic levels. Their determination requires sophisticated methods, expensive facilities and instruments, well-trained personnel and expensive chemical reagents. Ideally, real-time monitoring using rapid detection methods should be applied to detect possible contamination along the food chain in order to prevent human exposure. Sensor technology may be promising in this respect. This review gives the state of the art for detecting possible contamination with dioxins and DL-PCBs along the food chain of animal-source foods. The main detection methods applied (i.e., high resolution gas-chromatography combined with high resolution mass-spectrometry (HRGC/HRMS) and the chemical activated luciferase gene expression method (CALUX bioassay)), each have their limitations. Biosensors for detecting dioxins and related compounds, although still under development, show potential to overcome these limitations. Immunosensors and biomimetic-based biosensors potentially offer increased selectivity and sensitivity for dioxin and DL-PCB detection, while whole cell-based biosensors present interpretable biological results. The main shortcoming of current biosensors, however, is their detection level: this may be insufficient as limits for dioxins and DL-PCBs for food and feedstuffs are in pg per gram level. In addition, these contaminants are normally present in fat, a difficult matrix for biosensor detection. Therefore, simple and efficient extraction and clean-up procedures are required which may enable biosensors to detect dioxins and DL-PCBs contamination along the food chain.

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Enhancing the response of CALUX and CAFLUX cell bioassays for quantitative detection of dioxin-like compounds

Cited by 29 publications

References 17 publications

Development of Species-Specific Ah Receptor-Responsive Third Generation CALUX Cell Lines with Enhanced Responsiveness and Improved Detection Limits

Development of Species-Specific Ah Receptor-Responsive Third Generation CALUX Cell Lines with Enhanced Responsiveness and Improved Detection Limits

Environmental Application of Reporter-Genes Based Biosensors for Chemical Contamination Screening

The Need and Potential of Biosensors to Detect Dioxins and Dioxin-Like Polychlorinated Biphenyls along the Milk, Eggs and Meat Food Chain

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