Development of a monoclonal antibody recognizing benzo[c]phenanthrenediol epoxide-DNA adducts: application to immunohistochemical detection of DNA damage.

A monoclonal antibody was developed against (+/-)-anti-benzo[c]phenanthrenediol epoxidemodified DNA, and sensitivity and specificity were determined by competitive enzyme-linked immuno-sorbent assay (ELISA). Antibody 10F9 has 50% inhibition in the ELISA at 50 fmol of B[c]PhDE-DNA adducts. There was weak cross-reactivity with DNA modified by (+/-)-anti-benzo[a]pyrenediol epoxide (50% inhibition at 150 pmol). Testing of oligonucleotides containing either (+)- or (-)-trans-anti-B[c]PhDE-adenine adducts indicated similar recognition of both stereoisomers. A quantitative immunoperoxidase technique with antibody 10F9 was developed using 10T1/2 cells treated with B[c]PhDE then piloted on exfoliated oral cells from five smokers and five nonsmokers. Mean staining in smokers (184 +/- 11) was 1.64-fold higher than in nonsmokers (112 +/- 9, p < 0.0001). This antibody should be useful for the detection and quantitation of B[c]PhDE-DNA adducts in cell culture and animal studies and in humans with environmental or occupational exposure to polycyclic aromatic hydrocarbons.