Immunological detection and quantitation of DNA adducts formed by 4-aminoazobenzene species in vivo.
Antibodies to 3-methoxy-4-aminoazobenzene (3-MeO-AAB) and 2-methoxy-4-aminoazobenzene (2-MeO-AAB) DNA adducts were raised in rabbits against in vitro-adducted DNA samples. The enzyme-linked immunosorbent assay (ELISA) was used to determine the sensitivity and specificity of these antibodies. They proved highly specific for the modified DNA used as the immunogen, but cross-reacted with each other. Moreover, they showed cross reactivity with DNA modified by 4-(o-tolylazo)-o-toluidine, but not by other carcinogens, such as 4-aminobiphenyl or 4-nitroquinoline 1-oxide. The 50% inhibition level of antibody binding in the competitive ELISA was at 10-20 fmol of modified base per assay (equivalent to 1-2 adducts per 10(6) bases). Immunohistochemical staining indicated that these antibodies bind specifically to nuclear components of the liver in rats given either 3-MeO-AAB or 2-MeO-AAB at the dose of 50 mg/kg body weight.
Kojima
M
M
Laboratory of Biochemistry, Aichi Cancer Center Research Institute, Nagoya.
Morita
T
T
Shirai
T
T
Degawa
M
M
Tada
M
M
D000818:Animals; D000906:Antibodies / Q000276:immunology; D001667:Binding, Competitive; D002273:Carcinogens / Q000378:metabolism*; D004247:DNA / Q000302:isolation & purification / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D007150:Immunohistochemistry; D007274:Injections, Intraperitoneal; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051381:Rats; D011916:Rats, Inbred F344; D010128:p-Aminoazobenzene / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology
D000906:Antibodies; D002273:Carcinogens; C053767:2-methoxy-4-aminoazobenzene; D010128:p-Aminoazobenzene; C044436:3-methoxy-4-aminoazobenzene; D004247:DNA