Selective, Facile, Quantitative Detection of Oxidized Abasic Lesions in DNA
Abstract (Set) A variety of structural modifications (lesions) are produced in DNA when the biopolymer is exposed to oxidative stress. Lesions can be genotoxic and/or cytotoxic, and are useful biomarkers. Selective, quantitative detection of lesions is useful for rapidly determining the effects of DNA damaging agents. We have developed a simple method for selectively quantifying two oxidized abasic lesions at femtomole levels. The abasic lesions are detected using a biotinylated reagent synthesized in our laboratory. Quantification is carried out via fluorescence using horseradish peroxidase and Amplex RediI (Invitrogen). Description (Set) Proposed Use (Set) Currently, mass spectrometry is the most common method for quantifying DNA lesions. More specifically, this is typically accomplished by digesting the DNA to monomeric components, followed by LC/MS or GC/MS analysis. LC/MS is currently the method of choice. GC/MS requires additional derivatization. In addition, quantification is often carried out using isotopically labeled material as an internal standard, which limits user access because the labeled lesion requires chemical synthesis and is often unavailable to typical practitioners. Finally, mass spectrometry is a sensitive method but the instruments are expensive and difficult to use. This invention presents a simple, inexpensive reagent that enables one to quantify how much intermediate is produced using a routine fluorescence assay.
Greenberg, Marc M. ,Dhar, Shanta
Type of Offer:
« More Diagnostic Patents