32P-Postlabelling: A Sensitive Technique for the Detection of DNA Adducts

Abstract

The ³²P-postlabelling assay has emerged as a major method to detect DNA adducts induced by structurally diverse carcinogens and other DNA lesions. The technique comprises enzymatic degradation of DNA to 3'-monodeoxynucleotides, enrichment of adducts, 5'-³²P-labelling, adduct separation by TLC (or HPLC), and detection and quantitation of adducts. The review concerns individual ³²P-postlabelling techniques (standard procedure, enrichment methods) as well as the applications of the method to different DNA modifications. Recent reports on the application of the ³²P-postlabelling technique in human risk assessment using carcinogen-DNA adducts as molecular biomarkers are discussed and a critical evaluation of the assays is presented.