title: Immunoaffinity concentration of human lung DNA adducts using an anti-benzo[a]pyrene-diol-epoxide-DNA antibody. Analysis by 32P-postlabelling or ELISA.
abstract: DNA, isolated from 15 human lung autopsy samples, was examined for the presence of polycyclic aromatic hydrocarbon (PAH) DNA adducts. Using the nuclease P1 modification of the 32P-postlabelling technique, between 1 and 12 adducts/10(8) nucleotides were detected prior to immunoconcentration. Autoradiograms from most of the samples revealed a diagonal smear of radioactivity consistent with complex mixture (cigarette smoking) DNA damage. The DNA samples were digested to oligonucleotides, made single-stranded and subsequently applied to immunoaffinity columns containing immobilised anti-benzo[a]pyrene (B(a)P)-7,8-diol-9,10-epoxide (BPDE) DNA polyclonal rabbit antibody. The material remaining bound to the column, in addition to that passing through, was analysed using both ELISA and 32P-postlabelling techniques. Column-bound adducts comprised between 0% and 78% of any particular sample. Immunoconcentration, followed by 32P-postlabelling of the material which had been bound to the column, revealed the presence of a number of discrete adduct spots in autoradiograms of the more heavily adducted samples. Sample DNA not retained by the columns was also analysed; the chromatographic pattern obtained was a dense zone of radioactive material migrating from the origin. This evidence suggests that the composition of PAH-DNA adducts found in human lung samples exhibits wide inter-individual variation.
authors.affiliation: Jack Birch Unit for Environmental Carcinogenesis, Department of Biology, University of York, Heslington, UK.
authors.forename: M M
authors.initials: MM
authors.lastname: King
authors.forename: J
authors.initials: J
authors.lastname: Cuzick
authors.forename: D
authors.initials: D
authors.lastname: Jenkins
authors.forename: M N
authors.initials: MN
authors.lastname: Routledge
authors.forename: R C
authors.initials: RC
authors.lastname: Garner
mesh_terms: D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D000906:Antibodies; D002846:Chromatography, Affinity; D004247:DNA; D018736:DNA Adducts; D004249:DNA Damage; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008168:Lung / Q000737:chemistry* / Q000187:drug effects; D010761:Phosphorus Radioisotopes; D011083:Polycyclic Compounds / Q000032:analysis*
chemical_list: D000906:Antibodies; D018736:DNA Adducts; D010761:Phosphorus Radioisotopes; D011083:Polycyclic Compounds; C043931:benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D004247:DNA