Patent ID: 7148071
Filing Date: 2006-12-12
Classification: G01N,Y10S,Y10T

Abstract:
1. A method of detecting a binding event involving a test protein with a test ligand, the method comprising: (a) providing an unpurified test protein; (b) providing a test ligand; (c) contacting the test ligand with the unpurified test protein to form a first test mixture; (d) contacting the first test mixture with an exchange buffer comprising a denaturant and deuterium to form a second test mixture, the exchange buffer having a denaturant concentration; (e) contacting the second test mixture with a mass spectrometry matrix medium; (f) determining a change in mass of the test protein of the second test mixture resulting from hydrogen-deuterium exchange by mass spectrometry; (g) varying the denaturant concentration of the exchange buffer; (h) repeating steps (a)–(g) a number of times to generate a denaturation curve of the test protein; and (i) analyzing the change in mass of the test protein resulting from hydrogen-deuterium exchange as a function of denaturant concentration in the presence of the test ligand as compared to a change in mass of the test protein resulting from hydrogen-deuterium exchange as a function of denaturant concentration in the absence of the test ligand, wherein a difference in the changes in mass of the test protein as a function of denaturant concentration is indicative of a binding event involving the test protein and the test ligand.