High-throughput metabolic genotoxicity screening with a fluidic microwell chip and electrochemiluminescence

Wasalathanthri, Dhanuka P.; Malla, Spundana; Bist, Itti; Tang, Chi Kwong; Faria, Ronaldo C.; Rusling, James F.

doi:10.1039/c3lc50698c

Cited by 29 publications

(64 citation statements)

References 56 publications

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“…Hydrophobic barriers or wells can also be used to locate the drops, and the resulting LbL film, in specific regions of a flat surface. xxiv …”

Section: Basic Film Fabrication Methodologymentioning

confidence: 99%

“…xxiv An ideal representation of the LbL films is illustrated in Figure 10. The RuPVP catalyst cation (Scheme 1) is responsible for light generation by ECL when it is electrochemically activated and reacts with guanines in DNA.…”

Section: Multifunctional Enzyme/dna Films For Metabolic Toxicity Smentioning

confidence: 99%

“…LC-MS/MS analysis of products of magnetic bead reactors coated with the same enzyme/DNA films as in the arrays resulted in product ion spectra of dG-BPDE and dA-BPDE that confirmed formation of these adducts when cyt P450 and EH were on the beads. xxiv …”

Section: Multifunctional Enzyme/dna Films For Metabolic Toxicity Smentioning

confidence: 99%

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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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“…Hydrophobic barriers or wells can also be used to locate the drops, and the resulting LbL film, in specific regions of a flat surface. xxiv …”

Section: Basic Film Fabrication Methodologymentioning

confidence: 99%

Section: Multifunctional Enzyme/dna Films For Metabolic Toxicity Smentioning

confidence: 99%

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Thin multicomponent films for functional enzyme devices and bioreactor particles

2014

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“…A high-throughput ECL chip with 64 microwells capable of holding 1 lL droplets were integrated into a microfluidic system for measuring DNA damage caused by metabolites of benzo[a]pyrene [95]. In addition, a low-cost microfluidic paper electrochemical device (lPED) equipped with RuPVP/DNA/enzyme spots was used to screening the genotoxic compounds in water, food, and smoke [96,97].…”

Section: Genotoxicity Screeningmentioning

confidence: 99%

Imaging Analysis Based on Electrogenerated Chemiluminescence

et al. 2017

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Electrochemiluminescence (ECL), also called electrogenerated chemiluminescence, is luminescence that is produced by chemical reactions triggered by electrochemical method. ECL imaging is a novel imaging technique, which can simultaneously provide two kinds of signals of both electrochemistry and optical image. Over the past two decades, ECL imaging has been not only a powerful tool for investigating the fundamental scientific questions such as ECL mechanisms and reaction kinetics, but also a versatile analytical technique for detection of a wide range of analytes including small molecules, DNA, proteins, and cells. In the first part of this review, we briefly describe the reaction mechanisms of ECL generation. Then the review focuses on the research progress on the ECL imaging approach. It is basically introduced from the following five aspects: (i) visualization of electroactive sites on surfaces, (ii) imaging analysis at the single-bead and single-cell levels, (iii) array bioanalysis, (iv) multi-color ECL imaging, and (v) paper chip based on ECL imaging. Finally, some perspectives and future directions in this active research area are presented.

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“…5,37,38 Our most advanced ECL array features a 64-microwell reactor chip and are shown in Figure 2. 43 A computer printed pattern of hydrophobic ink is heat transferred to a conductive pyrolytic graphite sheet to form 64 microwells about 10–15 nm deep that are filled with RuPVP/enzyme/DNA films. Reaction media containing the test chemical flows into the reactor where metabolites are generated that can react with DNA.…”

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confidence: 99%

Modern approaches to chemical toxicity screening

Hvastkovs¹,

Rusling

2017

Current Opinion in Electrochemistry

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Chemical toxicity has a serious impact on public health, and toxicity failures of drug candidates drive up drug development costs. Many in vitro bioassays exist for toxicity screening, and newer versions of these tend to be high throughput or high content assays, some of which rely on electrochemical detection. Toxicity very often results from metabolites of the chemicals we are exposed to, so it is important that assays feature metabolic conversion. Combining bioassays, computational predictions, and accurate chemical pathway elucidation presents our best chance for reliable toxicity prediction. Employing electrochemical and electrochemiluminescent approaches, cell-free microfluidic arrays can measure relative rates of formation of DNA-metabolite adduct formation (a measure of genotoxicity) as well as DNA oxidation levels resulting from enzyme-generated metabolites. Enzymes for several organ types can be studied simultaneously. These arrays can be used to identify the most reactive metabolites, and subsequent mechanistic details can then be investigated with high throughput LC-HPLC using enzyme/DNA-coated magnetic beads.

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High-throughput metabolic genotoxicity screening with a fluidic microwell chip and electrochemiluminescence

Cited by 29 publications

References 56 publications

Thin multicomponent films for functional enzyme devices and bioreactor particles

Thin multicomponent films for functional enzyme devices and bioreactor particles

Imaging Analysis Based on Electrogenerated Chemiluminescence

Modern approaches to chemical toxicity screening

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