High-Throughput Miniaturization of Cytochrome P450 Inhibition Screening Using the Echo Liquid Handler
5 Nov 2012Cytochrome 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.
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Echo® 555 Liquid Handler
Beckman Coulter Life SciencesEcho® 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