Method and system for determination of molecular interaction parameters

1. A method of determining binding interaction 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, exposing the solid support surface to a plurality of fluid volumes containing different known concentrations of the binding partner to permit association of binding partner to the immobilized ligand,

b) 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

c) determining the kinetic parameters by globally fitting a predetermined binding interaction model for the interaction between the binding partner and the immobilized ligand to the collected binding data.

2. The method according to claim 1 , comprising a step a2) of exposing the solid support surface a fluid volume free from binding partner to permit dissociation of binding partner from the ligand.

3. The method of claim 1 , wherein steps a) to b) are repeated at least once with the same or another sequence of binding partner concentrations, and all the collected binding data are fitted simultaneously to the kinetic model in step c).

4. The method of claim 1 , wherein sequential binding data obtained from steps a) to b) are combined with at least one set of binding data for a single concentration of binding partner, and the combined binding data are fitted in step c) to the kinetic model.

5. The method of claim 2 , wherein step a2) is performed after flowing the last fluid volume containing binding partner over the surface.

6. The method of claim 2 , wherein step a2) is performed between two fluid volumes containing binding partner.

7. The method of claim 2 , wherein each volume containing binding partner is followed by a fluid volume free from binding partner.

8. The method of claim 7 , wherein one of the fluid volumes free from binding partner is exposed to the solid support surface for a longer time than the other fluid volumes free from binding partner.

9. The method of claim 8 , wherein the last fluid volume free from binding partner is exposed to the solid support surface for a longer time than the other fluid volumes free from binding partner.

10. The method of claim 2 , wherein the plurality of fluid volumes containing binding partner include fluid volumes with the same concentration of binding partner.

11. The method of claim 2 , wherein step a) is stopped when an increased concentration of binding partner results in a predetermined marginal increase in the amount of bound binding partner at the sensor surface.

12. The method of claim 2 , wherein the kinetic model is a two-compartment model.

13. The method of claim 2 , wherein the kinetic model is adapted to an interaction mechanism selected from 1:1 binding, parallel reactions, competing reactions, conformation change and bivalent analyte.

14. The method of claim 2 , wherein step a) is performed at at least one other density of ligand at the solid support surface.

15. The method of claim 2 , wherein the exposure of the fluid volumes to the surface is performed by flowing said volume over the solid support surface.

16. The method of claim 15 , wherein the flow over the solid support surface is maintained essentially constant.

17. The method of claim 2 , wherein the solid support surface is a sensor surface.

18. The method of claim 2 , wherein the detection at the solid support surface is based on evanescent wave sensing.

19. The method of claim 17 , wherein the detection at the solid support surface is based on surface plasmon resonance (SPR).

20. The method of claim 2 , wherein the detection at the solid support surface is based on detection of a label.

21. The method of claim 1 , wherein step c) is computer-implemented.