Surface plasmon resonance methods
First Claim
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1. A method comprising:
- a) measuring by SPR a first extent of interaction between a first ligand and a substrate in the presence of a second ligand, wherein the substrate possesses a first chemically reactive group and the second ligand possesses a second chemically reactive group and the second ligand is capable of forming a reversibly covalent substrate-ligand complex with the substrate, and wherein the first ligand or the substrate is immobilized to a surface, b) measuring by SPR a second extent of interaction between the first ligand and the substrate in the absence of the second ligand, and c) comparing the first extent of interaction with the second extent of interaction.
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Abstract
The present invention relates to the use of SPR methods to obtain information relating to molecular binding events. In one aspect of the present invention, methods are provided for determining whether a binding interaction is competitive or cooperative. In another aspect of the present invention, methods are provided for obtaining information where the binding interaction includes a formation of a reversible covalent bond.
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17 Claims
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1. A method comprising:
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a) measuring by SPR a first extent of interaction between a first ligand and a substrate in the presence of a second ligand, wherein the substrate possesses a first chemically reactive group and the second ligand possesses a second chemically reactive group and the second ligand is capable of forming a reversibly covalent substrate-ligand complex with the substrate, and wherein the first ligand or the substrate is immobilized to a surface, b) measuring by SPR a second extent of interaction between the first ligand and the substrate in the absence of the second ligand, and c) comparing the first extent of interaction with the second extent of interaction. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 13, 14, 17)
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12. A method comprising:
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a) measuring by SPR a first extent of interaction between a first ligand and a substrate-thiolate in the presence of a second ligand, wherein the substrate-thiolate possesses a first thiol group, and the second ligand possesses a second thiol group, and the substrate-thiolate and the second ligand are capable of forming a substrate-SS—
ligand complex, and wherein the first ligand or the substrate-thiolate is immobilized to a surface,b) measuring by SPR a second extent of interaction between the first ligand and the substrate-thiolate in the absence of the second ligand, and c) comparing the first extent of interaction with the second extent of interaction.
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15. A method comprising:
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a) measuring by SPR a calibration corresponding to an extent of formation of a reversible covalent bond between a substrate and a ligand in the presence of a reversing agent at a first concentration wherein the first concentration is sufficient to disrupt the reversible covalent bond and wherein the substrate or the ligand is immobilized, b) measuring by SPR a plurality of test values wherein each value corresponds to an extent of formation of the reversible covalent bond between the substrate and the ligand at a concentration of a reversing agent that is less than the first concentration, and c) correlating the extent of formation of the reversible covalent bond as a function of the concentration of the reversing agent. - View Dependent Claims (16)
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Specification