(1366-C) Real Time Cellular Thermal Shift Assay (RT-CETSA) Instrument Development

Abstract: The advantages of cellular thermal shift assay with real time monitoring have been previously described by scientists at the National Center for Advancing Translational Sciences (NCATS). Adding the real time thermal challenge component to the traditional CETSA method significantly increases throughput, efficiency, and data quality, leading to improved identification of ligand-target engagement in cells which is essential in hit-calling and the assessment of small molecule mechanism of action. A cleverly adapted prototype instrument that couples a real-time PCR device with a CCD camera for luminescence detection highlighted the potential of the method and displayed fundamental shortcomings. While the NCATS developed prototype and RT-CETSA assay negate the need for individual sample preparation at each temperature and other substantial optimization time sinks, it still required significant manual intervention and validation throughout an experiment. Subsequently, the team’s demand to use the prototype could not be met due to the vast amount of time it took to operate and manipulate the “Frankenstein” device. Establishing the utility of this successful prototype led NCATS to develop an instrument capable of meeting the needs for RT-CETSA experimentation. NCATS partnered with Arimation Robotics, an experienced custom automation/machine builder, to design and construct a new benchtop system to meet the required machine functions to run RT-CETSA experiments in an efficient, controlled, and safe manner, making this crucial technology accessible.