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8329192 
Book/Book Chapter 
Dna damage, repair, and genome instability (including affinity techniques) 
Wang, H; Lu, M; Dever, B; Shen, S; Le, XC 
2012 
Elsevier Inc. 
Comprehensive Sampling and Sample Preparation 
3 
231-260 
English 
Human DNA is often exposed to a variety of endogenous and exogenous agents that may induce a wide range of abnormal DNA modifications or change the patterns and levels of normal DNA modification such as the methylation of cytosine. The critical role of DNA modifications in cancer development makes it essential to develop highly sensitive and specific assays for qualitative and quantitative detection of exquisite changes in DNA modifications. By taking advantage of the high sensitivity of laser-induced fluorescence (LIF) detection, the high specificity of immuno-recognition, and the high efficiency of capillary electrophoresis (CE) separation, rapid and ultrasensitive CE-LIF immunoassays have been developed for analysis of DNA modification levels. The LOD for benzo(α)pyrene diol epoxide-DNA adducts is about 6.6 × 10-21 mol, which is 5400 times lower than that achieved using the nuclease P1-enhanced 32P-postlabeling assay. A number of methodologies have been established, including synthesis and characterization of adducted DNA fluorescent probes, interaction of DNA adducts and antibodies, stabilization of trace DNA adducts-antibody complexes, and DNA-driven focusing. These methodological studies and innovations contribute greatly to the improvement in the limit of detection and reproducibility. The developed methods have been applied in the mechanistic study of DNA repair and co-carcinogenesis, and are potentially useful in human biomonitoring and studies in health effects of carcinogens and co-carcinogens. © 2012 Elsevier Inc. All rights reserved. 
Biomarker; Capillary electrophoresis; DNA adduct; Immunoassay; Laser-induced fluorescence