A method of determining kinetic parameters for a reversible molecular interaction between a ligand immobilised 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 immobilised ligand, - flowing a plurality of fluid volumes containing different known concentrations (C 1 to C n) of the binding partner over the solid support surface to permit association of binding partner to the immobilised ligand. - flowing over the solid support surface a fluid volume free from binding partner to permit dissociation of binding partner from the ligand, b) monitoring using surface plasmon resonance biosensor during step a) - 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, and - for each fluid volume, the time point when injected (tOn) and when stopped (tOff), and c) determining the kinetic parameters by fitting globally a predetermined kinetic model for the interaction between the binding partner and the immobilised ligand to the collected binding data, which model allows for mass transport limitation at the solid support surface, wherein the global fitting is based on the time dependent concentration of the binding partner with respect to the time points when each fluid volume was injected (tOn) and stopped (tOff), and the density of ligand at the solid support surface. a) sequentially, without intermediate regeneration or renewal of the immobilised ligand, - flowing a plurality of fluid volumes containing different known concentrations (C 1 to C n) of the binding partner over the solid support surface to permit association of binding partner to the immobilised ligand. - flowing over the solid support surface a fluid volume free from binding partner to permit dissociation of binding partner from the ligand, - flowing a plurality of fluid volumes containing different known concentrations (C 1 to C n) of the binding partner over the solid support surface to permit association of binding partner to the immobilised ligand. - flowing over the solid support surface a fluid volume free from binding partner to permit dissociation of binding partner from the ligand, b) monitoring using surface plasmon resonance biosensor during step a) - 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, and - for each fluid volume, the time point when injected (tOn) and when stopped (tOff), and - 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, and - for each fluid volume, the time point when injected (tOn) and when stopped (tOff), and c) determining the kinetic parameters by fitting globally a predetermined kinetic model for the interaction between the binding partner and the immobilised ligand to the collected binding data, which model allows for mass transport limitation at the solid support surface, wherein the global fitting is based on the time dependent concentration of the binding partner with respect to the time points when each fluid volume was injected (tOn) and stopped (tOff), and the density of ligand at the solid support surface.