Analytical measuring and evaluation method for molecular interactions

摘要

The invention relates to an analytical measuring and evaluation method for determining the interaction parameters between an analyte and a ligand, preferably in a biosensor. According to the inventive method, the concentration of the analyte is gradually changed at defined intervals ti and the initial association or dissociation rates or association and dissociation rate constants are determined. The invention further relates to a device for carrying out the inventive method.

主权项

1. An analytical measurement and evaluation method for determining kinetic constants for molecular interactions between a first binding partner L (ligand) and a second binding partner A (analyte) forming a complex LA, wherein the first binding partner L is immobilized on a sensor surface of a biosensor, the sensor surface is contacted with a solution containing the second binding partner A, and binding of the second binding partner A to the immobilized binding partner L on the sensor surface is detected over time providing measurement values, the method comprising the following steps:
raising or lowering several times an analyte concentration cti(A) of the second binding partner A in a stepwise fashion and before an equilibrium interaction state for the respective analyte concentration cti(A) has been reached, and without any regeneration of the sensor surface between each raising or lowering of the analyte concentration; and determining one or more kinetic constants of the interaction from the measurement values.