ThermoFluor 384 Johnson and Johnson Miniaturized Fluorescence Based Thermal Shift Analyzer

The Johnson and Johnson / 3-Dimensional Pharmaceuticals ThermoFluor® 384 system performs miniaturized fluorescence based thermal shift assays for the high throughput screening of compound libraries. The ThermoFluor® screening strategy has several advantages over the more conventional HTS methodologies, the most important of which is that it relies on a general physical process common to most, if not all, drug targets - thermal unfolding of biomolecules and their ligand bound complexes. The ThermoFluor® instrument measures dye fluorescence in the presence of a protein. As the protein unfolds, its hydrophobic parts are exposed which creates an environment that causes the dye to fluoresce. The more the protein unfolds, the greater the intensity of the fluorescence.

In this manner, the protein's melting transition can be measured. More importantly for the ThermoFluor® screening strategy, the effects of ligand binding on the protein's melting transitions can be measured.

The ThermoFluor® 384 system provides a flexible and modular fluorescence assay system with an automated robotic microplate loading system to perform this function. The basic ThermoFluor® 384 is capable of sequentially loading and unloading up to twenty (20) 384-well thermal cycler microplates. The ThermoFluor® system works in conjunction with a personal computer running Total Control for Windows™ and 3-Dimensional Pharmaceuticals's ThermoFluor® Acquire 3.0 control software under Microsoft Windows™ NT.

The robotic microplate handler (PlateCrane™ by Hudson Control Group) moves sample plates from an input storage magazine to the imaging area of the instrument. As the microplate is removed from the input magazine, a bar code reader reads the plate's bar code identification number. Once the microplate is in the imaging area, the ThermoFluor® control software initiates a ThermoFluor® experiment on the microplate. When the experiment is completed, the robotic handler then retrieves the plate from the imaging area and places it into an output storage magazine. This process is repeated for up to 20 microplates in a single session. The PlateCrane™ robot and plate storage magazines (stacks) are optimized to handle a specified brand and model 384 well PCR plates.

Once the data have been collected, the 3-Dimensional Pharmaceuticals's ThermoFluor® analysis software is used to analyze the data to determine melting temperatures and identify hits.