Compound Inhibitor Profiling using the I.DOT

In this study the I.DOT Liquid Handler, a low-volume, non-contact liquid handling device was used for the performance of the ADP-Glo™ Kinase Assay, a commonly used assay for compound inhibitor profiling. The ADP-Glo™ Kinase Assay measures ADP formed from a kinase reaction, where a reactive phosphate group from ATP is transferred onto a target protein, thereby resulting in a remaining ADP molecule. This ADP is then converted into ATP, which is used to generate light in a luciferase reaction allowing quantification of the amount of phosphate groups transferred by the kinase reaction. Therefore, the assay is well suited for measuring the effects of chemical compounds on the activity of many purified kinases, making it ideal for primary screening as well as kinase selectivity profiling.

In these experiments we used the I.DOT to dispense different reagent addition steps or the whole assay. Two different enzymes and fourteen different compounds were used. The results were then compared to the workflow using another common low volume liquid handler and to hand pipetting. The experiments showed that the I.DOT is dispensing the ADP-Glo™ Kinase Assay with comparable results much faster and with less dead volume compared to the other liquid handler. Furthermore, using the I.DOT reduces the number of laboratory devices used for the assay, as well as reducing consumption of consumables and reagents. This ultimately leads to major cost savings.

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