||Detection of O6-Methyldeoxyguanosine in Human Placental DNA.
Foiles, P. G. ;
Miglietta, L. M. ;
Akerkar, S. A. ;
Everson, R. B. ;
Hecht, S. S. ;
||American Health Foundation, Inc., Valhalla, NY. Naylor Dana Inst. for Disease Prevention. ;National Inst. of Environmental Health Sciences, Research Triangle Park, NC. Epidemiology Branch.;Health Effects Research Lab., Research Triangle Park, NC.;National Cancer Inst., Bethesda, MD.
Deoxyribonucleic acids ;
Monoclonal antibodies ;
Antibody binding sites ;
High pressure liquid chromatography
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A monoclonal antibody specific for O(sup 6)-methyldeoxyguanosine (O(sup 6)-MedGuo) was developed. When used in a competitive ELISA, 50% inhibition of binding was achieved with 0.51 pmol O(sup 6)-MedGuo. When the competitive ELISA was coupled with HPLC, 2 mg of DNA could be analyzed giving a lower limit of detection of 0.5 micromol O(sup 6)-MedGuo per mol deoxyguanosine. The assay was used to test for O(sup 6)-MedGuo in DNA from placentas of smoking and non-smoking women. Two of 10 DNA samples from smoking women and 3 of 10 from non-smoking women had detectable levels of O(sup 6)-MedGuo. Levels ranged from 0.6 to 1.6 micromol O(sup 6)-MedGuo per mol deoxyguanosine. Levels of O(sup 6)-alkylguanine DNA alkyltransferase (AAT) protein were also measured. There was no apparent relationship between AAT and O(sup 6)-MedGuo levels in the 20 subjects, nor did mean AAT levels differ between the 2 groups. Although no apparent relationship between smoking history and O(sup 6)-MedGuo formation was found in the preliminary study, the report is the first of a structurally identified DNA adduct in human placenta.