Show simple item record

dc.contributor.authorWasalathanthri, Dhanuka P.
dc.contributor.authorMalla, Spundana
dc.contributor.authorBist, Itti
dc.contributor.authorTang, Chi K.
dc.contributor.authorFaria, Ronaldo C.
dc.contributor.authorRusling, James F.
dc.identifier.citationWasalathanthri, Dhanuka P. Malla, Spundana; Bist, Itti; Tang, Chi K.; Faria, Ronaldo C.; Rusling, James F. (2013). High-throughput metabolic genotoxicity screening with a fluidic microwell chip and electrochemiluminescence. Lab on a Chip 13 (23), 4554-4562
dc.description.abstractA high throughput electrochemiluminescent (ECL) chip was fabricated and integrated into a fluidic system for screening toxicity-related chemistry of drug and pollutant metabolites. The chip base is conductive pyrolytic graphite onto which are printed 64 microwells capable of holding one-mu L droplets. Films combining DNA, metabolic enzymes and an ECL-generating ruthenium metallopolymer ((RuPVP)-P-II) are fabricated in these microwells. The system runs metabolic enzyme reactions, and subsequently detects DNA damage caused by reactive metabolites. The performance of the chip was tested by measuring DNA damage caused by metabolites of the well-known procarcinogen benzo[a]pyrene (B[a]P). Liver microsomes and cytochrome P450 (cyt P450) enzymes were used with and without epoxide hydrolase (EH), a conjugative enzyme required for multi-enzyme bioactivation of B[a]P. DNA adduct formation was confirmed by determining specific DNA-metabolite adducts using similar films of DNA/enzyme on magnetic bead biocolloid reactors, hydrolyzing the DNA, and analyzing by capillary liquid chromatography-mass spectrometry (CapLC-MS/MS). The fluidic chip was also used to measure IC50-values of inhibitors of cyt P450s. All results show good correlation with reported enzyme activity and inhibition assays.
dc.publisherRoyal Society of Chemistry (RSC)
dc.relation.ispartofLab on a Chip
dc.rightsAttribution-NonCommercial-NoDerivs 3.0 Ireland
dc.subjecthuman liver-microsomes
dc.subjecthuman cytochrome-p450 enzymes
dc.subjecttandem mass-spectrometry
dc.subjectreactive metabolites
dc.subjectelectrogenerated chemiluminescence
dc.subjectbiocolloid reactors
dc.subjectsoft lithography
dc.titleHigh-throughput metabolic genotoxicity screening with a fluidic microwell chip and electrochemiluminescence

Files in this item


This item appears in the following Collection(s)

Show simple item record

Attribution-NonCommercial-NoDerivs 3.0 Ireland
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivs 3.0 Ireland