Use of fluorescence correlation spectroscopy to identify compounds that bind to target species under isothermal denaturing conditions

A high throughput screening method for identifying a test compound that binds to a target species, the method comprising:

• incubating a plurality of test mixtures under isothermal denaturing conditions, each test mixture comprising at least one test compound, at least one reporter molecule, and at least one target species, wherein the isothermal denaturing conditions are effective to cause at least a portion of the target species to denature to a measurable extent;
• detecting a denaturation signal of each target species in the presence of the at least one test compound by monitoring the diffusion properties of the target molecule using fluorescence correlation spectroscopy;
• and
• comparing the denaturation signal of each target species in the presence of at least one test compound with a denaturation signal of the same target species in the absence of the at least one test compound under the same isothermal denaturing conditions.

A high throughput screening method for identifying a test compound that binds to a protein, the method comprising:

• incubating a plurality of test mixtures under isothermal denaturing conditions, each test mixture comprising at least one test compound, at least one reporter molecule, and at least one protein, wherein the isothermal denaturing conditions are effective to cause at least a portion of the protein to denature to a measurable extent;
• detecting a denaturation signal of each protein in the presence of the at least one test compound by monitoring the difflusion properties of the target molecule using fluorescence correlation spectroscopy;
• and
• comparing the denaturation signal of each protein in the presence of at least one test compound with a denaturation signal of the same protein in the absence of the at least one test compound under the same isothermal denaturing conditions.