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

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

Research output: Contribution to journalArticle

23 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1010-1016
Number of pages7
JournalScience China Chemistry
Volume53
Issue number5
DOIs
StatePublished - May 2010

Fingerprint

Dioxins
Bioassay
Luciferases
Genes
Chemical detection
Cell signaling
Firefly Luciferases
Cell culture
Modulators
Assays
Screening
Cells
Proteins

Keywords

  • Bioassay
  • CAFLUX
  • CALUX
  • Dioxin
  • Reporter gene

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Enhancing the response of CALUX and CAFLUX cell bioassays for quantitative detection of dioxin-like compounds. / Zhao, Bin; Baston, David S.; Khan, Elaine; Sorrentino, Claudio; Denison, Michael S.

In: Science China Chemistry, Vol. 53, No. 5, 05.2010, p. 1010-1016.

Research output: Contribution to journalArticle

Zhao, Bin ; Baston, David S. ; Khan, Elaine ; Sorrentino, Claudio ; Denison, Michael S. / Enhancing the response of CALUX and CAFLUX cell bioassays for quantitative detection of dioxin-like compounds. In: Science China Chemistry. 2010 ; Vol. 53, No. 5. pp. 1010-1016.
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