Our Technology

Gyrasol’s Electron Transfer Quenching Platform

Gyrasol Technologies’s platform offers important advantages over other fluorecence-based assays: the activities of kinases, phosphatases, proteases and phosphodiesterases can be measured and quantified directly, with one uniform sensor.  The sensor works with various substrates, including peptides, lipids, oligonucleotides and cyclic nucleotides, that can be labeled with fluors of any spectral property. This is the only assay that can be mulitplexed, and run at physiological substrate (1-100 uM) and ATP  (100 uM to 2 mM) concentrations with purified enzymes or lysates in endpoint or kinetic modes. The platform is so sensitive that enzyme activity can be detected in as little as 200 ng of crude lysate. There is no need for antibodies, radioactivity or specialized equipment/filters.  The presence or absence of substrate (blue, green and red; Figure 1) phosphoryl groups is detected by the change in fluorescence of the fluor-labeled substrate when bound by the Sensor (yellow star) and  directly correlates to the level of substrate conversion.

The Gyrasol Technologies Sensor 01 is able to bind to phosphorylated substrates associated to any fluor and is optimal for multiplexed reactions and for applications where compound interferences are a concern. The Gyrasol Technologies Sensor 02 associates to phosphonates at pH > 9.0 and provides superior sensitivity of detection by harnessing the high quantum yield (0.9) of fluorescein at basic pH.

Advantages of the Gyrasol Platform:

• Homogeneous, direct detection formatted for HTS
• Adaptable to physiological substrate (1-100 μM) and ATP concentrations (up to 2 mM)
• Uses standard instrumentation
• No requirement for spectral overlap
• Adaptable to a variety of enzymes including phosphodiesterases, protein kinases, lipid kinases, phosphatases and proteases
• Multiplexible
• No radioactivity or costly antibodies
• Linear, quantifiable dose response
• Stable detection and no lot-to-lot variation