PURPOSE: Disclosed is a method for ranking the affinity of each of a multiplicity of different molecules for a target molecule which is capable of denaturing due to a thermal change. CONSTITUTION: The method comprises contacting the target molecule with one molecule of the multiplicity of different molecules in each of a multiplicity of containers; simultaneously heating the multiplicity of containers, measuring in each of the containers a physical change associated with the thermal denaturation of the target molecule resulting from the heating in each of the containers; generating a thermal denaturation curve for the target molecule as a function of temperature for each of the containers and determining a midpoint temperature (Tm) therefrom; comparing the Tm of each of the thermal denaturation curves with the Tm of a thermal denaturation curve obtained for the target molecule in the absence of any of the molecules in the multiplicity of different molecules; and ranking the affinities of the multiplicity of different molecules according to the change in Tm of each of the thermal denaturation curves.
MICRO-PLATE THERMAL SHIFT ASSAY AND APPARATUS FOR LIGAND DEVELOPMENT AND MULTI-VARIABLE PROTEIN CHEMISTRY OPTIMIZATION