The dedicated high-throughput screening reader PHERAstar FSX speeds up miniaturized analyses in 1536 well plates
Drug discovery screening carried out in microplates is used to screen extensive chemical libraries for compounds that show a specific activity or property. At this stage, after a successfully assay optimization, the screening assay has to be measured as fast as possible. This demands a very precise and robust instrumentation.
The following features of the PHERAstar® FSX have been developed to shorten the reading time as much as possible:
- Measurement in 1536-well and 3456-well plate formats
- Simultaneous detection of two emission signals as it is occurring for fluorescence polarization assays, BRET, FRET and TR-FRET assays as well as for multiplexing AlphaLISA®/AlphaPlex™ assays
- Use of a high-frequency laser in flying mode (1 flash) as excitation source in TR-FRET assays, such as HTRF®, LANCE® or LanthaScreen®
Assay miniaturization into 1536- or 3456-well microplates is one possibility to reduce costs in drug discovery screenings. To successfully run assays in these formats, liquids must be handled with extreme accuracy and the small volumes require highly sensitive detection. Liquid handlers such as the Echo® 555 from Labcyte fulfill these requirements.
In HTS, assay and instrument performance is expressed by the Z´ value. A value of > 0.5 should be reached at least, values of > 0.7 stand for excellent performance.
Fig. 1: Signal to Blank (S/B), Z´value and read times for a Histone-BRD4 interaction assay measured on the PHERAstar FSX. The use of the TRF laser reduces read times, increases S/B, while keeping a very high Z´value.
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Cellular and biochemical HTRF assays measured in 1536-well microplates
Increasing throughput with dual emission AlphaLISA-AlphaPlex assay and Simultaneous Dual Emission detection
Miniaturization of an HTRF methyltransferase assay that detects histone modifying activity
Searching for novel kinase inhibitors with the PHERAstar® at BIENTA laboratory in Kyiv, Ukraine. HTS group screened more than 600,000 compounds in kinase assays using ATP depletion assay, measuring luminescence in a coupled luciferase reaction.
PHERAstar plate reader is routinely used in our laboratory as it provides an accurate signal, high throughput and sensitivity, as well as stable temperature control required for luminescent kinase assays.