Detection of DNA damaging agents using layer-by-layer assembly

The layer-by-layer (LBL) electrostatic deposition method was used to fabricate multilayer films of pentaerythritol-based metallodendrimer with Ru-II terpyridine subunits (RuDen) that has a positive charge and ds-DNA (ds, double-stranded) that has a negative charge due to its phosphate backbone. Evidence of assembly was obtained by fabrication of (DNA vertical bar RuDen)(n) on quartz treated with poly(diallyldimethylammonium chloride), PDDA. The absorbance at 263 nm varied linearly with it in the range 1-6. For electrochemical monitoring of damage by styrene oxide, the assembly was on a glassy carbon electrode that was coated with a monolayer of aminobenzoic acid. The measurement was based upon the RuDen-catalyzed oxidation of sites, e.g. guanine, that are exposed when ds-DNA is damaged. The peak current at 1.07 V versus Ag vertical bar AgCl in square wave voltammetry increases with incubation time for 30 min. The process was also monitored by the shift in the spectrum of a long period grating (LPG) fiber coated with (DNA vertical bar RuDen)(5). A typical shift, which is due to changes in the refractive index of the coating, was 0.3 and 1.8 nm for 5 and 30 min exposures, respectively, using an algorithm that can measure a shift of 10(-4) nm. (c) 2004 Elsevier B.V. All rights reserved.