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.
Zhang
Y J
YJ
Division of Environmental Health Sciences/Cancer Center, Columbia University, New York, New York 10032, USA.
Geacintov
N E
NE
Santella
R M
RM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D002460:Cell Line; D018736:DNA Adducts / Q000737:chemistry* / Q000276:immunology; D004249:DNA Damage; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D010616:Phenanthrenes / Q000737:chemistry* / Q000276:immunology; D012907:Smoking / Q000473:pathology
D000911:Antibodies, Monoclonal; D018736:DNA Adducts; D010616:Phenanthrenes