High-Throughput Miniaturization of Cytochrome P450 Inhibition Screening Using the Echo Liquid Handler

5 Nov 2012

Cytochrome P450 enzymes are the largest group of drug-metabolizing enzymes and their potential interactions with drug candidates could lead to idiosyncratic toxicity. CYP3A4 is the most abundant cytochrome P450 enzyme in the liver and plays a major role in metabolizing xenobiotics. The Promega P450-Glo™ CYP3A4 Assay (Luciferin-IPA) offers a sensitive, specific and high-throughput luminescence assay for the examination of CYP3A4 inhibition. In this application note the Echo® 555 liquid handler from Labcyte was used for the miniaturization and optimization of the P450-Glo™ Assay with pooled Human Liver Microsomes (HLM). The transfer of enzymes, buffers, microsomes, inhibitors and substrates in small volumes to provide a sensitive and robust assay is demonstrated. Comparable IC50 values for four known CYP3A4 inhibitors are presented.

Echo® 555 Liquid Handler

Beckman Coulter Life Sciences

Echo® 555 liquid handler - the world's fastest acoustic dispensing platform for life science applications. The Echo® 555 liquid handler is the world’s fastest acoustic liquid handling platform for life science applications. Laboratories that demand very high throughput rely upon the Echo 555 liquid handler for round-the-clock trouble-free operation to enable the fastest discovery.The Echo 555 doubles the throughput of the Echo 550 and provides transfers from as many as 660,000 wells per day. Like the Echo 520 and Echo 550 liquid handlers, the Echo 555 uses "touchless" ADE transfer technology. ADE uses sound energy to transfer 2.5 nL droplets from Echo-qualified 384- and 1536-well source plates into 96-, 384-, 1536- and 3456-well destination plates.This unique touchless technology eliminates consumable tip costs, washing, and cross-contamination. Unsurpassed volumetric precision (≤ 8% CV) and accuracy (≤ 10% CV) ensure superior results. Independent studies have shown that Echo liquid handlers eliminate false negatives due to sample loss on tip surfaces in low-concentration transfers. Like the Echo 550 liquid handler, the Echo 555 liquid handler reports both fluid volume and DMSO hydration.Echo® 555 Liquid Handler Features: Tipless transfer, resulting in dramatic cost savings Accurate, precise movement of nanoliter-scale quantities, enabling assay miniaturization and further reducing costs Elimination of cross-contamination, carryover or leachates Better results through direct dilution of compounds, an improvement over serial dilution methods Improved information about sample integrity and improved transfer quality through measurement of DMSO hydration level Applications: Compound management Compound screening siRNA screening Biochemical assays Cell-based assays PCR Genotyping Gene expression

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High-Throughput ScreeningHigh-throughput screening (HTS) is an automated drug discovery technique for identification of active compounds against a compound library. Use HTS readers and integrated assay preparation / analysis workstations to screen your compounds. Identify active compounds against various HTS libraries, including membranes, proteins and peptides and HTS cell lines. Find the best high-throughput screening products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Pipettes and DispensersPipettes and dispensers are essential lab equipment used to dispense measured volumes of liquid accurately. Explore a range of manual and electronic pipettes, including single and multichannel, serological and volumetric pipettes, as well as a range of pipette tips. Bottle-top dispensers are also available for simple and fast solvent dispensing. Find the best pipettes & dispensers in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Microplate Readers / DetectorsMicroplate readers are used to automate the detection and analysis of labeled or label-free components in microplates during assays or live-cell monitoring. Microplate readers are generally distinguished by their mode of detection. Types include absorbance, luminescence, fluorescence intensity, fluorescence polarization, TRF / FRET and multimode microplate readers. Microplate readers deliver a high throughput of samples by reading multiple wells simultaneously, with the 96-well format the most commonly used. As a result, microplate readers are often used in the drug discovery, bioassays, research and pharmaceutical industries for screening applications. Microplate loading can also be automated, with robotic microplate stackers to increase throughput. Find the best microplate readers in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.ADME-ToxicologyADME-toxicology (ADME-Tox) studies are used in pharmacology and pharmacokinetics to assess the activity/toxicity of drugs <i>in vivo</i> or <i>in vitro</i>. Find bioassays for absorption, distribution, metabolism, and excretion of drug molecules including cytotoxicity, transporter/permeability, metabolism and activity assays as well as hepatocytes and cell lines for ADME. Find the best ADME-toxicology products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Cell-Based AssaysCell-based assays are used to monitor the presence, quantity and activities of a desired cellular analyte including drug molecules or biomarkers. This can reveal information on cell health (apoptosis, cytotoxicity, viability and proliferation assays), cell metabolism, cell migration and cell signaling mechanisms. Find the best cell-based assay products, kits and equipment with our peer reviewed product directory: compare products, check customer reviews and receiving pricing direct from manufacturers.Robotic WorkstationsRobotic workstations are automated platforms enabling the user to perform high-throughput, multi-application processes, from sample preparation tasks such as PCR set-up, DNA purification and NGS library preparation, to integrated sample prep and analysis workstations. Automation makes processes consistent, fast, precise and fully walk-away. Find the best robotic workstations in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Automated Liquid HandlingAutomated liquid handling equipment is used to mix, dilute and dispense allotted volumes of liquid into microplates and other vessels automatically. The robotic, liquid handling arms can dispense from single channel to 3456 multichannel as well as operate nanoliter dispensing, enabling high throughput of samples. Find the best automated liquid handlers in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.MetabolomicsMetabolomics is the study of small metabolites (the intermediates and products of metabolism). It involves the identification and quantification of cellular metabolites using analytical technologies such as GC, HPLC, NMR, and LC/MS.CYP3A4CYPCytochrome P450 AssaysLiquid Handling WorkstationLiquid handling workstations automate the process of sample preparation, improving accuracy and efficiency. These systems are essential for high-throughput applications in biological, chemical, and pharmaceutical research, reducing human error and increasing reproducibility. Workstations come equipped with robotic arms, pipetting tools, and other accessories to streamline lab workflows. Explore liquid handling workstations in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Drug DevelopmentDrug development refers to the process of bringing a new drug to market.
High-Throughput Miniaturization of Cytochrome P450 Inhibition Screening Using the Echo Liquid Handler