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Biomarkers for Drug Discovery: Important Aspects of in vitro Assay Design for HTS and HCS Bioassays

William A. Irwin ; GlaxoSmithKline Istraživački Centar, Zagreb, Hrvatska
Dubravko Jelić ; GlaxoSmithKline Istraživački Centar, Zagreb, Hrvatska
Roberto Antolović ; GlaxoSmithKline Istraživački Centar, Zagreb, Hrvatska

Puni tekst: engleski pdf 131 Kb

str. 23-30

preuzimanja: 1.588



High throughput screening (HTS) is the foundation of current drug discovery to assay drug candidates for toxicity and biological effects (i.e. off-target and on-target responses, respectively). HTS is typically based on measuring thousands of drug candidates per day with a single endpoint assay on a limited number of doses or even a single dose of compound. The assays can be either based on absorbance or fluorescence measurements (i.e. Alamar Blue, MTT, Fluo-4 for calcium, etc.). Conversely, high content screening (HCS) is based on measuring a limited number of drugs per day, but measuring up to eight different assays simultaneously with multiple drug doses and even kinetic measurements. HCS assays typically are based on fluorescence microscopy and automatic image analysis algorithms. With HCS technology, tedious and time consuming assays can now be automated (i.e. nuclear size, micronucleus assay, lysosomal mass, mitochondrial membrane potential, neurite outgrowth, etc.). Multi-channel FACS (fluorescence activated cell sorting) can also be considered to be »high content« analysis. The purpose of this essay is to review important aspects of in vitro assay design common to both HTS and HCS screening technologies.

Ključne riječi

HTS, HCS, assay, in vitro, models, optimization, biomarkers

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