Development of Monoclonal Antibodies to the Malondialdehyde−Deoxyguanosine Adduct, Pyrimidopurinone1
| Autor: | Norma H. Mahle, Naomi Eliezer, Lawrence J. Marnett, Cynthia L. Sevilla, Shawn M. O'Hara, Ryan J. Miller, Munetaka Nokubo, Adam Uzieblo, Carol A. Rouzer |
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| Rok vydání: | 1997 |
| Předmět: | |
| Zdroj: | Chemical Research in Toxicology. 10:172-180 |
| ISSN: | 1520-5010 0893-228X |
| DOI: | 10.1021/tx960120d |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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