featuring the Echo Acoustic Technology
TITLES and AUTHORS
The advent of a new technology often brings far-reaching benefits well beyond the initial perception of its key advantages. Acoustic dispensing, for example, with the ability to have highly controlled droplet transfers at 2.5nL increments with exquisite precision and accuracy, does much more than save money by eliminating pipets. It enables new work flows, including those in which multiple pharmaceutical firms have seen dramatic differences in the true success or failure of compounds. This article shows how the use of acoustic dispensing and direct dilution leads to the creation of better compound structures and the potential for faster development of new drugs.
The quality of data generated within primary screening is dependant on three things: the quality of the compounds, the quality of the biology in the test well and the quality of data analysis.Traditionally compounds are diluted from concentrated stocks using aqueous diluents before testing in primary assays. This reduces the amount of organic solvent in the assay while lowering the compound concentration. However, reducing the concentration of solvent can allow the compound to drop out of solution. Acoustic Droplet Ejection (ADE) technology allows for nanolitre volumes of reagents to be transferred into assay wells. This removes the requirement for intermediate dilution of stocks and ensures that the compounds are held in optimum solvent conditions prior to assay. To investigate the impact of ADE on primary screening, 10,000 samples from the AZ screening collection were dispensed using ADE and tested in a primary screening assay. This data was then directly compared with the original primary screening data generated using aqueous dilutions and traditional contact dispensers.
With the aim of fuelling open-source, translational, early-stage drug discovery activities, the results of the recently completed antimycobacterial phenotypic screening campaign against Mycobacterium bovis BCG with hit confirmation in M. tuberculosis H37Rv were made publicly accessible. A set of 177 potent noncytotoxic H37Rv hits was identified and will be made available to maximize the potential impact of the compounds toward a chemical genetics/proteomics exercise, while at the same time providing a plethora of potential starting points for new synthetic lead-generation activities. Two additional drug-discovery- relevant datasets are included: a) a drug-like property analysis reflecting the latest lead-like guidelines and b) an early lead-generation package of the most promising hits within the clusters identified.
Early ADMET assessment is expected to not only improve the overall quality of drug candidates, but also shorten the drug discovery and development process. Time-dependent CYP450 inhibition and metabolic stability assays are critical in identifying drug candidates that may have undesirable drug-drug interactions or sub-optimal pharmacokinetic properties. The cost of profiling these assays during early drug development with large numbers of compounds however, can dramatically increase the expense of drug discovery. One objective for pharmaceutical companies is to find less expensive, more reliable and higher throughput ADMET methods that can be moved upstream. Here, we present a miniaturization method using the Echo® liquid handler to evaluate ADMET characteristics. The Echo liquid handler uses non-contact acoustic energy to transfer low volumes compounds and reagents. This new method not only generates high quality and reliable IC50 and intrinsic clearance values, but also significantly reduces reagent costs.
Recently identified associations between variants of cancer genes and drug resistance increase the value for comprehensive drug sensitivity and resistance testing in combination with molecular profiling of cancer cells. The measure of sample sensitivity or resistance to a drug requires high throughput screening of engineered cancer cell lines or samples directly from affected patients against combinations of anti-cancer therapeutics. Results are compared with genetic profiles in an attempt to determine the more effective treatment. Advancements in next generation sequencing and qPCR technologies persuade many research organizations to increase effort in these areas. In doing so, researchers immediately recognize screening efficiency as a critical factor to accurate and reproducible drug sensitivity and resistance testing. This application note discusses the implementation of miniaturized drug sensitivity and resistance testing, at the Institute of Molecular Medicine in Finland (FIMM), with assay-ready plates produced by the Echo liquid handler.
Commonly used cancer treatments are not always effective against malignantly transformed cells. Drug repositioning, the application of existing drugs not used currently against a given cancer, is a way to develop new treatments. In this film you can see how drug sensitivity testing, the process by which patient cells are examined experimentally for response in activity to various drugs, is done at the Technology Centre of FIMM, the Institute for Molecular Medicine, Finland.
Single nucleotide polymorphism (SNP) genotyping of maize samples is commonly performed in agricultural science to aide marker assisted selection, study heterosis and a variety of other biological behaviors. Its widespread adoption in agricultural science has been challenged with increasing reagent costs and labor intensive multi-step processes. Acoustic non-contact liquid handling using the Echo 525 liquid handler offers unique advantages to traditional processes by incorporating a tip-less solution to deliver reagents precisely and accurately. Assay miniaturization is enabled with high accuracy and precision at volumes as low as 25 nL. This study utilized the Echo 525 liquid handler to miniaturize a KASP genotyping assay for maize at a throughput meeting the demands of most high throughput production processes. The results demonstrate miniaturization with the Echo 525 liquid handler without compromising throughput targets.
Early assessment of ADMET (Absorption, Distribution, Metabolism, Excretion and Toxicity) properties plays a critical role in the optimization and prediction of pharmacokinetic behaviors of new chemical entities. Among the most important ADME assays is the in vitro drug metabolic stability
assay. This assay evaluates the susceptibility of compounds to biotransformation or intrinsic clearance. Because an increasing number of compounds require metabolic stability evaluation, a high-throughput and cost-effective method is desirable during early drug discovery. In this application note, we discuss how the Echo liquid handler provides a platform for conducting a miniaturized high-throughput metabolic stability assay. The Echo liquid handler precisely and accurately transfers nanoliter volumes with acoustic energy in a completely contact-free manner. Thus high quality data can be obtained with a small fraction of the volume required by tip-based liquid handling methods.
Acoustic transfer with the Echo liquid handler simplifies the small scale crystallography process by transferring reagents precisely at 2.5 nL increments. Using Dynamic Fluid Analysis™, the Echo liquid handler can adjust transfer settings on the fly on a well-by-well basis. This allows a single fluid class setting to be utilized for a wide variety of viscous and osmotic fluids, greatly simplifying the liquid transfer process. We demonstrate an expanded ability to transfer a wide variety of reagents from various protein crystallography sparse matrix screens. To enable the potential for on the fly grid screens, we demonstrate the precise transfer of viscous reagents (including high molecular weight PEGs) and organic solvents (including MPD).
Selecting the most suitable liquid handling method can have a huge influence over your final results, particularly with miniaturised volumes. Acoustic liquid handling using the relatively new direct dilution technique may offer scientists greater accuracy.