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

Yu Jing Zhang, Nicholas 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

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DNA Adducts
Epoxy Compounds
DNA Damage
Monoclonal Antibodies
Immunosorbents
DNA
Antibodies
Assays
Enzyme-Linked Immunosorbent Assay
Stereoisomerism
Polycyclic Aromatic Hydrocarbons
Environmental Exposure
Adenine
Enzymes
Occupational Exposure
Immunoenzyme Techniques
Cell culture
Oligonucleotides
Animals
Cell Culture Techniques

ASJC Scopus subject areas

  • Chemistry(all)
  • Organic Chemistry
  • Health, Toxicology and Mutagenesis
  • Drug Discovery
  • Toxicology

Cite this

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title = "Development of a monoclonal antibody recognizing benzo[c]phenanthrenediol epoxide-DNA adducts: Application to immunohistochemical detection of DNA damage",
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.",
author = "Zhang, {Yu Jing} and Nicholas Geacintov and Santella, {Regina M.}",
year = "1997",
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AU - Santella, Regina M.

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N2 - 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.

AB - 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.

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