Monoclonal antibodies to 1-aminopyrene-DNA

L. L. Hsieh*, A. M. Jeffrey, R. M. Santella

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

14 Scopus citations

Abstract

Monoclonal antibodies were obtained after fusion of mouse P3X63-AG.8.653 myeloma cells with spleen cells isolated from BALB/cCr mice immunized with denatured DNA modified by 1-nitrosopyrene reduced with sodium ascorbate (AP-d-DNA) and complexed electrostatically to methylated bovine serum albumin. Ten stable hybridoma lines have been isolated and characterized by enzyme-linked immunosorbent assay (ELISA). They all recognize 1-aminopyrene (1-AP)modified DNA, but not free 1-nitropyrene or 1-aminopyrene. Antibody 11H2 is the most specific for AP-DNA showing no cross-reactivity with unmodified native DNA. It also recognizes 8-nitro-1-AP and 6-nitro-1-AP modified DNA. There was some low cross-reactivity with DNA modified by a benzo[a]pyrene diol epoxide and N-acetoxy-N-2-acetylaminofluorene. Competitive ELISA with antibody 11H2 reliably detected AP-DNA adducts formed when 1-nitropyrene was incubated with Salmonella typhimurium TA1538. By immunological methods, AP-DNA adducts were shown to be unstable to heat denaturation. This suggests that specific monoclonal antibodies to carcinogen-DNA adducts will be useful not only for detecting and quantitating carcinogen-DNA damage but also for probing adduct stability.

Original languageEnglish
Pages (from-to)1289-1293
Number of pages5
JournalCarcinogenesis
Volume6
Issue number9
DOIs
StatePublished - 09 1985
Externally publishedYes

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