Miniaturization of High-Throughput Epigenetic Methyltransferase Assays with Acoustic Liquid Handling

Edwards, Bonnie; Lesnick, John; Wang, Jing; Tang, Nga; Peters, Carl

doi:10.1177/2211068215610861

Cited by 10 publications

(8 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cells were homogenized in 50 mM Tris-HCl (pH 7.4) and centrifuged at 500× g for 10 min at 4 °C. Supernatant was utilized for the homogenous time-resolved fluorescence (HTRF) assay (Cisbio kit) [14] , [15] . Cell lysate, as an enzyme source, was reacted with the methyl donor SAM and incubated for 45 min at room temperature (RT).…”

Section: Methodsmentioning

confidence: 99%

Forskolin attenuates doxorubicin-induced accumulation of asymmetric dimethylarginine and s-adenosylhomocysteine via methyltransferase activity in leukemic monocytes

Ramachandran

Loganathan

Cheeran

et al. 2018

Leukemia Research Reports

View full text Add to dashboard Cite

Doxorubicin (DOX) is an antitumor drug, associated with cardiomyopathy. Strategies to address DOX-cardiomyopathy are scarce. Here, we identify the effect of forskolin (FSK) on DOX-induced-asymmetric-dimethylarginine (ADMA) accumulation in monocytoid cells. DOX-challenge led to i) augmented cytotoxicity, reactive-oxygen-species (ROS) production and methyltransferase-enzyme-activity identified as ADMA and s-adenosylhomocysteine (SAH) accumulation (SAH-A). However, except cytotoxicity, other DOX effects were decreased by metformin and FSK. FSK, did not alter the DOX-induced cytotoxic effect, but, decreased SAH-A by >50% and a combination of three drugs restored physiological methyltransferase-enzyme-activity. Together, protective effect of FSK against DOX-induced SAH-A is associated with mitigated methyltransferase-activity, a one-of-a-kind report.

show abstract

Section: Methodsmentioning

confidence: 99%

Forskolin attenuates doxorubicin-induced accumulation of asymmetric dimethylarginine and s-adenosylhomocysteine via methyltransferase activity in leukemic monocytes

Ramachandran

Loganathan

Cheeran

et al. 2018

Leukemia Research Reports

View full text Add to dashboard Cite

show abstract

“…One way to minimize the consumption of reagents when screening very large numbers of compounds is to miniaturize and parallelize an assay into 1536-well microtiter plates [50]. However, such miniaturization requires the addition of very small volumes of compound stock solutions and technologies such as the contactless acoustic dispenser from Labcyte Inc. makes this possible [51]. …”

Section: Assay Development High Throughput and High Content Screeninmentioning

confidence: 99%

Epigenetic assays for chemical biology and drug discovery

Gul

2017

Clin Epigenet

View full text Add to dashboard Cite

The implication of epigenetic abnormalities in many diseases and the approval of a number of compounds that modulate specific epigenetic targets in a therapeutically relevant manner in cancer specifically confirms that some of these targets are druggable by small molecules. Furthermore, a number of compounds are currently in clinical trials for other diseases including cardiovascular, neurological and metabolic disorders. Despite these advances, the approved treatments for cancer only extend progression-free survival for a relatively short time and being associated with significant side effects. The current clinical trials involving the next generation of epigenetic drugs may address the disadvantages of the currently approved epigenetic drugs.The identification of chemical starting points of many drugs often makes use of screening in vitro assays against libraries of synthetic or natural products. These assays can be biochemical (using purified protein) or cell-based (using for example, genetically modified, cancer cell lines or primary cells) and performed in microtiter plates, thus enabling a large number of samples to be tested. A considerable number of such assays are available to monitor epigenetic target activity, and this review provides an overview of drug discovery and chemical biology and describes assays that monitor activities of histone deacetylase, lysine-specific demethylase, histone methyltransferase, histone acetyltransferase and bromodomain. It is of critical importance that an appropriate assay is developed and comprehensively validated for a given drug target prior to screening in order to improve the probability of the compound progressing in the drug discovery value chain.

show abstract

“…The source plates used to transfer from were 384-well polypropylene microtiter plates. A variety of different well fluid volumes were used in the source plates (20,30,40, and 50 µL). For each source well volume studied, 20-drop transfers from 384 separate wells filled to that volume were measured, in order to obtain meaningful statistics.…”

Section: Dynamic Fluid Analysismentioning

confidence: 99%

Moving Liquids with Sound: The Physics of Acoustic Droplet Ejection for Robust Laboratory Automation in Life Sciences

Hadimioglu¹,

Stearns²,

Ellson³

2016

SLAS Technology

View full text Add to dashboard Cite

Liquid handling instruments for life science applications based on droplet formation with focused acoustic energy or acoustic droplet ejection (ADE) were introduced commercially more than a decade ago. While the idea of “moving liquids with sound” was known in the 20th century, the development of precise methods for acoustic dispensing to aliquot life science materials in the laboratory began in earnest in the 21st century with the adaptation of the controlled “drop on demand” acoustic transfer of droplets from high-density microplates for high-throughput screening (HTS) applications. Robust ADE implementations for life science applications achieve excellent accuracy and precision by using acoustics first to sense the liquid characteristics relevant for its transfer, and then to actuate transfer of the liquid with customized application of sound energy to the given well and well fluid in the microplate. This article provides an overview of the physics behind ADE and its central role in both acoustical and rheological aspects of robust implementation of ADE in the life science laboratory and its broad range of ejectable materials.

show abstract

Miniaturization of High-Throughput Epigenetic Methyltransferase Assays with Acoustic Liquid Handling

Cited by 10 publications

References 19 publications

Forskolin attenuates doxorubicin-induced accumulation of asymmetric dimethylarginine and s-adenosylhomocysteine via methyltransferase activity in leukemic monocytes

Forskolin attenuates doxorubicin-induced accumulation of asymmetric dimethylarginine and s-adenosylhomocysteine via methyltransferase activity in leukemic monocytes

Epigenetic assays for chemical biology and drug discovery

Moving Liquids with Sound: The Physics of Acoustic Droplet Ejection for Robust Laboratory Automation in Life Sciences

Contact Info

Product

Resources

About