Determination of coupling constants by the equilibrium shift method.

摘要

The present invention relates to a method for determining the binding constant between a substance or substance mixture and a target and to a kit for carrying out the method according to the invention. The basic functional principle of the method according to the invention is the determination of the binding constant of the substance to the target by shifting the binding equilibrium. In this regard, affinity for the target is determined by varying the concentration of the two targets, immobilized target and dissolved target, and shifting the binding equilibrium in the individual batches. In this regard, at least four samples are used, and according to the present invention, first and second samples of a substance comprising immobilized targets of different substance amounts and dissolved targets of the same substance quantity are incubated, Incubating a third and a fourth sample of a substance comprising an immobilized target of the same substance amount as the first and second samples and containing a soluble target of the same substance quantity, the solubility in the sample containers 3 and 4 The amount of target material differs from the amount of soluble target material in sample containers 1 and 2, and the sample container contains the same volume of liquid phase consisting of buffer, dissolved target, and material sample during incubation.