Evaluation of Electrochemiluminescent Metabolic Toxicity Screening Arrays Using a Multiple Compound Set

Pan, Shenmin; Zhao, Linlin; Schenkman, John B.; Rusling, James F.

doi:10.1021/ac200050n

Cited by 29 publications

(87 citation statements)

References 41 publications

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“…27 HLM induced faster DNA oxidation and adduction rates than RLM for all the aryl amines, in agreement with previous work using a non-microfluidic manual array. 18 Supersome 1A2 had the largest relative DNA oxidation and adduction rates for 4-ABP, AF, and NA, while for anisidine, the 2E1 isoform caused slightly more bioactivation for DNA oxidation and adduction than 1A2, consistent with other in vitro studies. 25,44 …”

Section: Resultssupporting

confidence: 88%

“…QCM frequency shifts were used to calculate the mass densities and nominal film thicknesses. 14,18,27 Films were nominally 22–48 nm thick depending on layer mass densities (Table S2 and S3). …”

Section: Resultsmentioning

confidence: 99%

“…Relative DNA damage and oxidation rates are proportional to the slopes of % ECL increase vs time 11 and were converted to relative turnover rates (R) by dividing the initial slopes (t −1 ) by the total amount of protein in each film ( μ g) and the amount of substrate used (mM). 18 …”

Section: Resultsmentioning

confidence: 99%

“…Line graphs showing correlations of relative DNA damage rates (R) from human liver microsomes found by using the ECL array with the reciprocal of rodent liver TD 50 values and comet assay values (OTM/C in AU*mM −1 ) from human urinary bladder cell lines for 4-ABP and NA 45 and human peripheral lymphocytes for AF. 18,46 (C) and (D) shows correlation with relative DNA oxidation rate and relative DNA adduction rates for 4-ABP, AF, anisidine, and NA.…”

Section: Figurementioning

confidence: 99%

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Evaluating Metabolite-Related DNA Oxidation and Adduct Damage from Aryl Amines Using a Microfluidic ECL Array

Bist¹,

Bhakta²,

Jiang³

et al. 2017

Anal. Chem.

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Damage to DNA from the metabolites of drugs and pollutants constitutes a major human toxicity pathway known as genotoxicity. Metabolites can react with metal ions and NADPH to oxidize DNA or participate in SN2 reactions to form covalently linked adducts with DNA bases. Guanines are the main DNA oxidation sites, and 8-oxo-7,8-dihydro-2-deoxyguanosine (8-oxodG) is the initial product. Here we describe a novel electrochemiluminescent (ECL) microwell array that produces metabolites from test compounds and measures relative rates of DNA oxidation and DNA adduct damage. In this new array, films of DNA, metabolic enzymes, and an ECL metallopolymer or complex assembled in microwells on a pyrolytic graphite wafer are housed in dual microfluidic chambers. As reactant solution passes over the wells, metabolites form and can react with DNA in the films to form DNA adducts. These adducts are detected by ECL from a RuPVP polymer that uses DNA as a coreactant. Aryl amines also combine with Cu2+ and NADPH to form reactive oxygen species (ROS) that oxidize DNA. The resulting 8-oxodG was detected selectively by ECL-generating bis(2,2′-bipyridine)-(4-(1,10-phenanthrolin-6-yl)-benzoic acid)Os(II). DNA/enzyme films on magnetic beads were oxidized similarly, and 8-oxodG determined by LC/MS/MS enabled array standardization. The array limit of detection for oxidation was 720 8-oxodG per 106 nucleobases. For a series of aryl amines, metabolite-generated DNA oxidation and adduct formation turnover rates from the array correlated very well with rodent 1/TD50 and Comet assay results.

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Section: Resultssupporting

confidence: 88%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Figurementioning

confidence: 99%

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Evaluating Metabolite-Related DNA Oxidation and Adduct Damage from Aryl Amines Using a Microfluidic ECL Array

Bist¹,

Bhakta²,

Jiang³

et al. 2017

Anal. Chem.

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“…xiv,lii,lv Both approaches provide DNA-damage rates that are correlated to one another, to cell-based DNA damage assays, and to animal genotoxicity metrics. lvi The arrays can be used to screen metabolites that react with DNA, then biocolloid reactors generate samples that are used to elucidate structures and measure formation rates of individual nucleobase adducts after hydrolyzing the DNA. The devices we have developed are inexpensive, high throughput, and can be employed at early development stages to contribute important toxicity-related reactivity and structure information for chemical product candidates.…”

Section: Multifunctional Enzyme/dna Films For Metabolic Toxicity Smentioning

confidence: 99%

Thin multicomponent films for functional enzyme devices and bioreactor particles

2014

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Complex functional films containing enzymes and other biomolecules are easily fabricated in nm-scale thicknesses by using layer-by-layer (LbL) methodologies first popularized by Lvov and Decher. In this review, we highlight the high level functional capabilities possible with LbL films of biomolecules based on our own research experiences. We first describe the basics of enzyme film fabrication by LbL alternate electrostatic adsorption, then discuss how to make functional enzyme-polyion films of remarkably high stability. Focusing on cytochrome P450s, we discuss films developed to electrochemically activate the natural catalytic cycle of these key metabolic enzymes. We then describe multifunctional, multicomponent DNA/enzyme/polyion films on arrays and particle surfaces for high throughput metabolic toxicity screening using electrochemiluminescence and LC-MS/MS. Using multicomponent LbL films, complex functionality for bioanalytical and biochemical purposes can be achieved that is difficult or impossible using conventional approaches.

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Electrochemiluminescence for Biomolecule Analysis

Guo,

Ding,

2024

Encyclopedia of Analytical Chemistry

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Electrochemiluminescence ( ECL ) is a light‐emitting process triggered by electrochemical reactions. Thanks to its low background, high sensitivity, wide dynamic range, and rapid response time, ECL has manifested itself as a leading signal transduction technique in biosensing and bioimaging. In recent years, much effort has been expended to extend the readout of ECL signal from the conventional intensity measurement to spectral and imaging analysis. ECL‐based spectroscopy ( ECLS ) and ECL‐based microscopy ( ECLM ) are thus established and became two of the most popular electroanalytical methods. Recent advances in ECL analysis not only provide insightful information on ECL reaction mechanisms but also improve the analytical performance of ECL bioassays, particularly for multiplexing capacity and spatiotemporal resolution. In this monograph, we shall briefly introduce the fundamentals of ECL, and then focus on the progress of ECL bioassays that mainly include immunoassays, nucleic acid detection, enzymatic assays, and single‐cell analysis. Finally, some challenges and perspectives of ECL bioassays are discussed and presented. We believe that there are plenty of opportunities to design new ECL bioassays with high sensitivity, high throughput, and high spatiotemporal resolution.

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Evaluation of Electrochemiluminescent Metabolic Toxicity Screening Arrays Using a Multiple Compound Set

Cited by 29 publications

References 41 publications

Evaluating Metabolite-Related DNA Oxidation and Adduct Damage from Aryl Amines Using a Microfluidic ECL Array

Evaluating Metabolite-Related DNA Oxidation and Adduct Damage from Aryl Amines Using a Microfluidic ECL Array

Thin multicomponent films for functional enzyme devices and bioreactor particles

Electrochemiluminescence for Biomolecule Analysis

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