Immunoelectrophoretic Detection of DNA Adducts in White Blood Cells

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University of Alberta (Canada), 2007 - Benzopyrene - 100 pages
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A fluorescence immunoassay with capillary electrophoresis and laser induced fluorescence (FIA/CE/LIF) technique was developed and tested suitable for the detection of benzo(a)pyrene-diol-epoxide (BPDE) DNA adducts in mononuclear white blood cells (MNCs). The developed method was tested for both in vitro and biomonitoring applications. Firstly, in vitro exposure of MNCs to both BPDE and buthionine sulfoximine, a glutathione depleting agent, resulted in an increase in adducts compared to BPDE exposure alone. In carrying out this experiment, we confirmed the important role of glutathione in BPDE inactivation in MNCs, and validated the use of FIA/CE/LIF for in vitro studies of BPDE-DNA adducts. To test the suitability for human biomonitoring, FIA/CE/LIF analysis was performed on MNCs of smokers. The adducts were detectable, however not quantifiable. Further improvements to sensitivity, as well as assessment of adducts in MNCs from other smokers, will help to identify whether FIA/CE/LIF is an appropriate technique for human biomonitoring of BPDE-DNA adducts.

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