Production and immunochemical characterization of a high-affinity monoclonal antibody specific for 7-(benzo[a]pyren-6-yl)guanine (BP-6-N7Gua), a depurinating dna adduct of benzo[a]pyrene

George P Casale, Eleanor G Rogan, Douglas Stack, Prabu Devanesan, Ercole Cavalieri

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Molecular dosimetry of depurinating DNA adducts of benzo[a]pyrene (BP) offers a promising new approach to determining risk of PAH-induced cancer. Depurinating adducts are the predominant form of BP-induced DNA damage, are excreted in urine and, importantly, are linked to cancer initiation. We have produced a monoclonal antibody (MAb) with high specific affinity for BP-6-N7Gua, a major depurinating DNA adduct of BP, and have developed a sensitive and specific competitive enzyme-linked immunosorbent assay (ELISA). The Isos (quantity producing 50% inhibition of MAb binding in the ELISA) of selected BP adducts and metabolites were determined. The results indicated 1) a high degree of discrimination between BP-6-N7Gua (Iso=750 finol) and BP (150=900,000 finol), 2) high affinity of the MAb for BP-6-N7Ade (I50=1,500 fmol), another major depurinating DNA adduct, and 3) specific structural requirements for MAb-adduct binding. In addition, the competitive ELISA provided an accurate determination of BP-6-N7Gua added to normal human urine, at a sensitivity of 200 fmol.

Original languageEnglish (US)
Pages (from-to)195-201
Number of pages7
JournalPolycyclic Aromatic Compounds
Volume10
Issue number1-4
DOIs
StatePublished - Jan 1 1996

Fingerprint

Monoclonal antibodies
Benzo(a)pyrene
Pyrene
Immunosorbents
Monoclonal Antibodies
Assays
Enzymes
DNA Adducts
Enzyme inhibition
Polycyclic aromatic hydrocarbons
Metabolites
Dosimetry
DNA
7-(benzo(a)pyren-6-yl)guanine

Keywords

  • Depurinating DNA adducts
  • ELISA
  • Monoclonal antibody
  • PAH

ASJC Scopus subject areas

  • Polymers and Plastics
  • Organic Chemistry
  • Materials Chemistry

Cite this

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title = "Production and immunochemical characterization of a high-affinity monoclonal antibody specific for 7-(benzo[a]pyren-6-yl)guanine (BP-6-N7Gua), a depurinating dna adduct of benzo[a]pyrene",
abstract = "Molecular dosimetry of depurinating DNA adducts of benzo[a]pyrene (BP) offers a promising new approach to determining risk of PAH-induced cancer. Depurinating adducts are the predominant form of BP-induced DNA damage, are excreted in urine and, importantly, are linked to cancer initiation. We have produced a monoclonal antibody (MAb) with high specific affinity for BP-6-N7Gua, a major depurinating DNA adduct of BP, and have developed a sensitive and specific competitive enzyme-linked immunosorbent assay (ELISA). The Isos (quantity producing 50{\%} inhibition of MAb binding in the ELISA) of selected BP adducts and metabolites were determined. The results indicated 1) a high degree of discrimination between BP-6-N7Gua (Iso=750 finol) and BP (150=900,000 finol), 2) high affinity of the MAb for BP-6-N7Ade (I50=1,500 fmol), another major depurinating DNA adduct, and 3) specific structural requirements for MAb-adduct binding. In addition, the competitive ELISA provided an accurate determination of BP-6-N7Gua added to normal human urine, at a sensitivity of 200 fmol.",
keywords = "Depurinating DNA adducts, ELISA, Monoclonal antibody, PAH",
author = "Casale, {George P} and Rogan, {Eleanor G} and Douglas Stack and Prabu Devanesan and Ercole Cavalieri",
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T1 - Production and immunochemical characterization of a high-affinity monoclonal antibody specific for 7-(benzo[a]pyren-6-yl)guanine (BP-6-N7Gua), a depurinating dna adduct of benzo[a]pyrene

AU - Casale, George P

AU - Rogan, Eleanor G

AU - Stack, Douglas

AU - Devanesan, Prabu

AU - Cavalieri, Ercole

PY - 1996/1/1

Y1 - 1996/1/1

N2 - Molecular dosimetry of depurinating DNA adducts of benzo[a]pyrene (BP) offers a promising new approach to determining risk of PAH-induced cancer. Depurinating adducts are the predominant form of BP-induced DNA damage, are excreted in urine and, importantly, are linked to cancer initiation. We have produced a monoclonal antibody (MAb) with high specific affinity for BP-6-N7Gua, a major depurinating DNA adduct of BP, and have developed a sensitive and specific competitive enzyme-linked immunosorbent assay (ELISA). The Isos (quantity producing 50% inhibition of MAb binding in the ELISA) of selected BP adducts and metabolites were determined. The results indicated 1) a high degree of discrimination between BP-6-N7Gua (Iso=750 finol) and BP (150=900,000 finol), 2) high affinity of the MAb for BP-6-N7Ade (I50=1,500 fmol), another major depurinating DNA adduct, and 3) specific structural requirements for MAb-adduct binding. In addition, the competitive ELISA provided an accurate determination of BP-6-N7Gua added to normal human urine, at a sensitivity of 200 fmol.

AB - Molecular dosimetry of depurinating DNA adducts of benzo[a]pyrene (BP) offers a promising new approach to determining risk of PAH-induced cancer. Depurinating adducts are the predominant form of BP-induced DNA damage, are excreted in urine and, importantly, are linked to cancer initiation. We have produced a monoclonal antibody (MAb) with high specific affinity for BP-6-N7Gua, a major depurinating DNA adduct of BP, and have developed a sensitive and specific competitive enzyme-linked immunosorbent assay (ELISA). The Isos (quantity producing 50% inhibition of MAb binding in the ELISA) of selected BP adducts and metabolites were determined. The results indicated 1) a high degree of discrimination between BP-6-N7Gua (Iso=750 finol) and BP (150=900,000 finol), 2) high affinity of the MAb for BP-6-N7Ade (I50=1,500 fmol), another major depurinating DNA adduct, and 3) specific structural requirements for MAb-adduct binding. In addition, the competitive ELISA provided an accurate determination of BP-6-N7Gua added to normal human urine, at a sensitivity of 200 fmol.

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