Patent ID: 7373255
Filing Date: 2008-05-13
Classification: B01L,G01N,Y10S

Abstract:
1. A method of determining kinetic parameters for a reversible molecular interaction between a ligand immobilized to a solid support surface and a binding partner to the ligand in solution, comprising the steps of: a) sequentially, without intermediate regeneration or renewal of the immobilized ligand, flowing a plurality of fluid volumes containing different known concentrations of the binding partner over the solid support surface to permit association of binding partner to the immobilized ligand, b) flowing over the solid support surface a fluid volume free from binding partner to permit dissociation of binding partner from the ligand, c) monitoring during steps a) and b) the momentary amount of binding partner bound to the solid support surface related to time and solution concentration of binding partner and collecting the binding data, wherein steps a) to c) are repeated at least once with the same or another sequence of binding partner concentrations, and d) determining the kinetic parameters by simultaneously fitting a predetermined kinetic model for the interaction between the binding partner and the immobilized ligand to all the collected binding data, which model allows for mass transport limitation at the solid support surface.