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

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  • Special Topic · Research Progress of Persistent Organic Pollutants
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Abstract

Reporter genes produce a protein product in transfected cells that can be easily measured in intact or lysed cells and they have been extensively used in numerous basic and applied research applications. Over the past 10 years, reporter gene assays have been widely accepted and used for analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related dioxin-like compounds in various types of matrices, such as biological, environmental, food and feed samples, given that high-resolution instrumental analysis techniques are impractical for large-scale screening analysis. The most sensitive cell-based reporter gene bioassay systems developed are the mechanism-based CALUX (Chemically Activated Luciferase Expression) and CAFLUX (Chemically Activated Fluorescent Expression) bioassays, which utilize recombinant cell lines containing stably transfected dioxin (AhR)-responsive firefly luciferase or enhanced green fluorescent protein (EGFP) reporter genes, respectively. While the current CALUX and CAFLUX bioassays are very sensitive, increasing their lower limit of sensitivity, magnitude of response and dynamic range for chemical detection would significantly increase their utility, particularly for those samples that contain low levels of dioxin-like HAHs (i.e., serum). In this study, we report that the addition of modulators of cell signaling pathways or modification of cell culture conditions results in significant improvement in the magnitude and overall responsiveness of the existing CALUX and CAFLUX cell bioassays.

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Authors and Affiliations

  1. State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085, China

    Bin Zhao

  2. Department of Environmental Toxicology, University of California, Davis, California, 95616, USA

    Bin Zhao & Michael S. Denison

  3. Laboratorium voor Analytische en Milieu Chemie, Vrije Universiteit Brussel, B-1050, Brussels, Belgium

    David S. Baston

  4. Pesticide Epidemiology Section, Pesticide and Environmental Toxicology Branch, Office of Environmental Health Hazard Assessment, Sacramento, California, 94612, USA

    Elaine Khan

  5. Department of Toxic Substances Control, Human and Ecological Risk Office, Brownfields and Environmental Restoration Program, Sacramento, California, 95812, USA

    Claudio Sorrentino

Authors
  1. Bin Zhao
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  2. David S. Baston
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  3. Elaine Khan
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  4. Claudio Sorrentino
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  5. Michael S. Denison
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Correspondence to Bin Zhao.

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Zhao, B., Baston, D.S., Khan, E. et al. Enhancing the response of CALUX and CAFLUX cell bioassays for quantitative detection of dioxin-like compounds. Sci. China Chem. 53, 1010–1016 (2010). https://doi.org/10.1007/s11426-010-0142-8

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  • DOI: https://doi.org/10.1007/s11426-010-0142-8

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