×

Method for determining the binding behavior of ligands which specifically bind to target molecules

  • US 20040232077A1
  • Filed: 11/15/2003
  • Published: 11/25/2004
  • Est. Priority Date: 05/23/2001
  • Status: Active Grant
First Claim
Patent Images

1. A method for determining the binding behavior of ligands which bind specifically to target molecules at at least one binding site, which method comprises the following steps:

  • (a) preparation of a first mixed phase wherein the ligands in a concentration K1 are contacted with markers, in a concentration K2, which bind specifically to the target molecules, and with the target molecules in a concentration K3, and of a second mixed phase wherein the markers are contacted with the target molecules, wherein after the contacting, the first and second mixed phases are incubated under identical conditions which permit a binding of the ligands and the markers to the target molecules, (b) separation of the bound markers GM1 contained in the first mixed phase from the first mixed phase, and of the bound markers GM2 contained in the second mixed phase from the second mixed phase, and (c1) determination of a concentration K4 of the unbound markers in the first mixed phase, and of a concentration K5 of the unbound markers in the second mixed phase, wherein a concentration K6 is determined from the concentration K5, which concentration K6 corresponds to the concentration of the unbound markers in the second mixed phase under the assumption that the markers in the concentration K2 have been contacted therein with the target molecules in the concentration K3, and determination of the binding behavior of the ligands from the ratio between the concentrations K6 and K4, or (c2) determination of a quantity M1 of the bound markers GM1 and of a quantity M2 of the bound markers GM2, wherein a quantity M3 is determined form the quantity M2, which amount M3 corresponds to the quantity of the bound markers in the second mixed phase under the assumption that the markers in the concentration K2 have been contacted therein with the target molecules in the concentration K3, and determination of the binding behavior of the ligands from the ratio between the quantities M3 and M1, wherein the markers are present in a native form and the determination of the concentrations K4 and K5 or of the quantities M2 and M1 is effected by means of mass spectrometry.

View all claims
  • 1 Assignment
Timeline View
Assignment View
    ×
    ×