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 Scopus citations

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
Publication statusPublished - Jan 1 1996

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Keywords

  • Depurinating DNA adducts
  • ELISA
  • Monoclonal antibody
  • PAH

ASJC Scopus subject areas

  • Polymers and Plastics
  • Organic Chemistry
  • Materials Chemistry

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