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, comprises 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,
monitoring 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 determining the kinetic
parameters by globally fitting a predetermined kinetic model for the
interaction between the binding partner and the immobilized ligand to the
collected binding data, which model allows for mass transport limitation
at the solid support surface. An analytical system for carrying out the
method, a computer program, a computer program product and a computer
system for performing the method are also disclosed.