title: Development of monoclonal antibodies to the malondialdehyde-deoxyguanosine adduct, pyrimidopurinone.
abstract: Malondialdehyde (MDA), an endogenous product of lipid peroxidation and prostaglandin biosynthesis, is mutagenic in bacterial and mammalian cells and carcinogenic in rats. In order to determine whether MDA-modified bases are formed in nucleic acids in vivo, sensitive immunoassays to detect MDA-DNA and MDA-RNA adducts are being developed in our laboratory. Murine monoclonal antibodies reactive with the MDA-deoxyguanosine adduct 3-beta-D-erythro-pentofuranosylpyrimido[1,2-alpha]purin-10(3H)-one (M1G-R) were prepared and characterized. Several MDA-modified nucleosides and deoxynucleosides and structural analogs were synthesized and characterized and were compared as competitive inhibitors in enzyme-linked immunosorbent assays (ELISAs). Less than 5 fmol of M1G in MDA-modified DNA was detected in a direct ELISA, and antibody binding to the modified DNA was competitively inhibited by free M1G-dR. DNA from Salmonella typhimurium treated with concentrations of MDA that induce reversion to histidine prototrophy was enzymatically digested, and M1G-dR was quantitated by competitive ELISA. Over a range of MDA concentrations from 10 to 40 mM, the level of M1G residues in bacterial DNA increased from 0.2 to 2.5/10(6) base pairs.
authors.affiliation: Proteins International, Rochester Hills, Michigan 48309, USA.
authors.forename: C L
authors.initials: CL
authors.lastname: Sevilla
authors.forename: N H
authors.initials: NH
authors.lastname: Mahle
authors.forename: N
authors.initials: N
authors.lastname: Eliezer
authors.forename: A
authors.initials: A
authors.lastname: Uzieblo
authors.forename: S M
authors.initials: SM
authors.lastname: O'Hara
authors.forename: M
authors.initials: M
authors.lastname: Nokubo
authors.forename: R
authors.initials: R
authors.lastname: Miller
authors.forename: C A
authors.initials: CA
authors.lastname: Rouzer
authors.forename: L J
authors.initials: LJ
authors.lastname: Marnett
mesh_terms: D000171:Acrolein / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry; D001666:Binding Sites, Antibody; D001667:Binding, Competitive / Q000276:immunology; D004247:DNA / Q000378:metabolism; D018736:DNA Adducts / Q000276:immunology* / Q000494:pharmacology; D004269:DNA, Bacterial / Q000032:analysis; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000276:immunology* / Q000494:pharmacology; D006825:Hybridomas / Q000737:chemistry; D006868:Hydrolysis; D008315:Malondialdehyde / Q000276:immunology* / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D012486:Salmonella typhimurium / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D018736:DNA Adducts; D004269:DNA, Bacterial; D008315:Malondialdehyde; D000171:Acrolein; C089188:3-(2'-deoxyribofuranosyl)pyrimido(1,2-a)purin-10(3H)-one; D004247:DNA; D003849:Deoxyguanosine