In Situ detection of DNA adducts formed in cultured cells by Benzo(a)pyrene diolepoxide (BPDE), with monoclonal antibodies specific for the BP‐Deoxyguanosine Adduct

Abstract

This paper describes the development of a method to assess exposure to benzo(a)pyrene (BP) by means of immunodetection of DNA adducts formed by the reactive BP‐diolepoxide (BPDE). The reaction product of BPDE with deoxyguanosine‐5'‐monophosphate was used as a hapten to raise antibodies in mice. Hybrid cells obtained by fusion of plasmacytoma cells with spleen cells from immunized mice, were screened for BP‐adduct‐specific antibody production in an enzyme‐linked immunosorbent assay (ELISA). Antibodies from a suitable clone were characterized in competitive immunoassays and used for in situ detection of BP‐DNA adduct formation in cultured fibroblasts treated with BPDE, by means of immunofluorescence microscopy. The adduct levels in these cells were determined with an independent method, viz. ³²P‐postlabelling.

The sensitivity reached with these methods may very well be adequate to apply these techniques in biomonitoring studies to assess occupational exposure of humans to BP.

Key words:

• Benzo(a)pyrene
• immunodetection
• DNA adducts