Patent ID: 8859295
Filing Date: 2014-10-14
Classification: G01N

Abstract:
1. A method of determining the dissociation constant (K d ) for a particular protein and ligand combination, comprising: providing a first solution comprising said protein with no ligand and a predetermined minimum amount of chemical denaturant; and a second solution comprising said protein, no ligand and a predetermined maximum amount of said chemical denaturant; mixing said first and second solution in a plurality of samples, each having said first and second solution in different proportions to create solutions having different molarities of said chemical denaturant; measuring an observable property of each of said plurality of samples; creating a denaturation graph based on said observable property and said molarity; determining the molarity at which a predetermined fraction of said protein molecules became denatured (F preparing a third solution comprising said protein and a ligand at a specified concentration ([L]) and an amount of chemical denaturant equal to said determined molarity; measuring said observable property of said third solution to determine a fraction of said protein molecules that became denatured in the presence of said ligand (F using said specified concentration ([L]), said predetermined fraction (F where A is defined as