Development of a monoclonal antibody recognizing benzo[c]phenanthrenediol epoxide-DNA adducts: Application to immunohistochemical detection of DNA damage

Yu Jing Zhang, Nicholas E. Geacintov, Regina M. Santella

Research output: Contribution to journalArticle

Abstract

A monoclonal antibody was developed against (±)-anti- benzo[c]phenanthrenediol epoxide-modified DNA, and sensitivity and specificity were determined by competitive enzyme-linked immunosorbent assay (ELISA). Antibody 10F9 has 50% inhibition in the ELISA at 50 fmol of B[c]PhDE-DNA adducts. There was weak cross-reactivity with DNA modified by (±)-antibenzo[a]pyrenediol epoxide (50% inhibition at 150 pmol). Testing of oligonucleotides containing either (+)- or (-)-trans-anti-B[c]PhDE-adenine adducts indicated similar recognition of both stereoisomers. A quantitative immunoperoxidase technique with antibody 10F9 was developed using 10T 1/4 cells treated with B[c]PhDE then piloted on exfoliated oral cells from five smokers and five nonsmokers. Mean staining in smokers (184 ± 11) was 1.64-fold higher than in nonsmokers (112 ± 9, p < 0.0001). This antibody should be useful for the detection and quantitation of B[c]PhDE-DNA adducts in cell culture and animal studies and in humans with environmental or occupational exposure to polycyclic aromatic hydrocarbons.

Original languageEnglish (US)
Pages (from-to)948-952
Number of pages5
JournalChemical research in toxicology
Volume10
Issue number9
DOIs
StatePublished - Sep 1997

ASJC Scopus subject areas

  • Toxicology

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