Antibodies against 7-methyldeoxygunaosine: Its detection in rat peripheral blood lymphocyte DNA and potential apllications to molecular epidemiology

P. Degan, R. Montesano, C. P. Wild

Research output: Contribution to journalArticle

Abstract

Polyclonal antibodies have been raised against the imidazole ring-open form of 7-methyldeoxyguanosine (7-mdGua). A combined high performance liquid chromatography/immunoassay method has been developed using these antibodies which provides a specific and sensitive way to quantitate 7-mdGua in DNA. Following enzyme hydrolysis and chromatographic purification of 7-mdGua, the adduct is quantitatively converted to the ring-open form and can be measured at levels as low as 0.05 pmol by immunoassay. With 1 mg of DNA a level below 1 adduct per 107 normal deoxynucleosides can be measured. Using DNA modified by radiolabeled carcinogens, a good correlation between 7-mdGua levels, as measured by immunoassay or radioactivity, was obtained. In rats treated with dimethylnitrosamine (0.4 and 1.0 mg/kg), both 7-mdGua and O6-methyldeoxyguanosine were detected in peripheral blood lymphocyte DNA. In addition the levels of both adducts at time points up to 48 h posttreatment were very similar to those seen in liver DNA from the same animals. The measurement of 7-mdGua, quantitatively the major methylation adduct, in small cell samples such as lymphocytes has great potential in determining the exposure of humans to environmental methylating agents such as nitrosamines.

Original languageEnglish
Pages (from-to)5065-5070
Number of pages6
JournalCancer Research
Volume48
Issue number18
Publication statusPublished - 1988

    Fingerprint

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this