Three Different Liquid Handling Systems to Achieve Flexibility and Scalability in Compound Screening against 400+ Protein Kinases

Abstract

AssayQuant Technologies has developed a chelation enhanced fluorescence (CHeF) assay that enables the screening of proprietary sensor peptides to monitor the activity of an individual kinases in a continuous assay format. AssayQuant leverages this technology in its KinSightTM profiling service – a kinome screening platform that can evaluate a single compound against a panel of greater than 400 kinase targets. To meet the increasing demands of these unique selectivity profiling services, the automation team at AssayQuant developed a robust, automated plating process that accounts for several factors: 400+ kinases currently in the profile, two different ATP conditions at which kinase assays are routinely performed (1mM and Km), and special assay conditions that specific kinases require for adequate enzymatic activity. The automation team considered throughput and reliability in an automation platform as well as instrument redundancy to guarantee that high-quality data collection, analysis, and reporting are completed with accuracy and efficiency.

Three automation systems were designed and implemented to accomplish these requirements. Inventory management and labware standardization was established to allow for uniform plating of all assay reagents (kinases, sensor peptides, compounds, and assay buffer components) on any liquid handler. The plating protocol was designed with cherry picking capabilities to enable additional flexibility and thus allow for assays to be run with ATP at 1 mM or at each individual kinase’s Km. Overall, the three systems implemented differ in deck capacity, throughput, and required user intervention, however, when combined they ensure redundancy and allow for increased overall throughput.

Key Takeaways

• KinSight Profiling screens compounds against up to 400 different wild type kinases per profiling panel. 34 assay plates per KinSight Profile Panel.
• The utilization of Master plates allows for KinSight to be performed at either 1mM ATP or Km.
• Each automated platform utilizes the same Master plates to generate the assay plates.
• Three automation systems enables flexibility and scalability with regards to turnaround time and capacity.
• Flexible and agnostic to screening contents. New kinases can be incorporated readily with minimal method modifications needed.
• Phosphatases or kinase mutant screening can be incorporated within the KinSight Panel or as a new service using the same methods.
• Custom Kinase panels employ the same strategy irrespective of the number of kinases in the custom panel.