MICROPLATE THERMAL SHIFT ASSAY AND APPARATUS FOR LIGAND DEVELOPMENT AND MULTI-VARIABLE PROTEIN CHEMISTRY OPTIMIZATION

摘要

The present invention is a method for ranking the affinity of each of amultiplicity of different molecules for a target moleculewhich is capable of denaturing due to a thermal change. The method comprisescontacting the target molecule with one molecule of themultiplicity of different molecules in each of a multiplicity of containers,simultaneously heating the multiplicity of containers, measuring ineach of the containers a physical change associated with the thermaldenaturation of the target molecule resulting from the heating in eachof the containers, generating a thermal denaturation curve for the targetmolecule as a function of temperature for each of the containers anddetermining a midpoint temperature (T m) therefrom, comparing the T m of eachof the thermal denaturation curves with the T m of a thermaldenaturation curve obtained for the target molecule in the absence of any ofthe molecules in the multiplicity of different molecules, andranking the affinities of the multiplicity of different molecules according tothe change in T m of each of the thermal denaturation curves.The present invention also provides an assay apparatus for that includes atemperature adjusting means for simultaneously heating a pluralityof samples, and a receiving means for receiving spectral emission from thesamples while the samples are being heated. In further aspectsof the invention, the receiving means can be configured to receive fluorescentemission, ultraviolet light, and visible light. The receivingmeans can be configured to receive spectral emission from the samples in avariety of ways, e.g., one sample at a time, simultaneously frommore than one sample, or simultaneously from all of the samples. Thetemperature adjusting means can be configured with a temperaturecontroller for changing temperature in accordance with a pre-determinedprofile.