Parallel analysis of molecular interactions
First Claim
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1. A method of detecting a molecular interaction, comprising the steps of:
- a) contacting an array with one or more target molecules, the array comprising a plurality of different affinity molecules in discrete domains, each domain having a predefined address in the array;
b) interrogating the array with a probe having a tip to create a profile of the array; and
c) evaluating the profile to detect an interaction between at least one affinity molecule and at least one target molecule.
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Abstract
Provided are methods of detecting molecular interactions using arrays and near field scanning probe techniques. Also provided are methods of characterizing binding interactions under defined reaction parameters, methods of determining antibody binding specificity, methods of selecting a substrate for an array of immobilized molecules and methods of determining molecular occupancy time with respect to binding interactions.
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Citations
34 Claims
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1. A method of detecting a molecular interaction, comprising the steps of:
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a) contacting an array with one or more target molecules, the array comprising a plurality of different affinity molecules in discrete domains, each domain having a predefined address in the array;
b) interrogating the array with a probe having a tip to create a profile of the array; and
c) evaluating the profile to detect an interaction between at least one affinity molecule and at least one target molecule. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method of determining antibody specificity comprising:
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a) contacting an array with an antigen, the array comprising a plurality of antibodies arranged in discrete domains, each of the domains having a predefined address in the array;
b) interrogating the array with a probe having a tip to create a profile of the array;
c) evaluating the profile to detect an antibody-antigen interaction in one or more of the domains; and
d) correlating the antibody-antigen interaction with antibody specificity. - View Dependent Claims (12, 13, 14, 15, 16)
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17. A method of determining antibody specificity comprising:
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a) contacting an array with at least one antibody, the array comprising a plurality of antigens arranged in discrete domains, each of the domains having a predefined address in the array;
b) interrogating the array with a probe having a tip to create a profile of the array;
c) evaluating the profile to detect an antibody-antigen interaction in one or more of the domains; and
d) correlating the antibody-antigen interaction with antibody specificity. - View Dependent Claims (18, 19, 20, 21, 22)
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23. A method of characterizing a molecular interaction comprising the steps of:
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a) contacting an array with one or more target molecules under defined reaction parameters, the array comprising a plurality of affinity molecules in discrete domains, each domain having a predefined address in the array;
b) interrogating the array with a probe having a tip to create a profile of the array;
c) evaluating the profile to detect an interaction between at least one affinity molecule and at least one target molecule in one or more domains; and
d) correlating the interaction with the binding conditions to characterize the molecular interaction. - View Dependent Claims (24, 25, 26, 27, 28)
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29. A method of selecting a substrate for an array of immobilized molecules comprising:
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a) contacting an array with at least one target molecule, the array comprising a plurality of substrates arranged in discrete domains and at least one affinity molecule disposed on the substrates in each of the domains;
b) interrogating the array with a probe having a tip to create a profile of the array;
c) evaluating the profile to detect a molecular interaction in one or more of the domains; and
d) selecting one or more of the substrates based on the profile. - View Dependent Claims (30, 31, 32, 33)
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34. A method of determining target occupancy time comprising:
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a) contacting an array with one or more target molecules, the array comprising a plurality of affinity molecules in discrete domains, each domain having a predefined address in the array;
b) interrogating the array with a probe having a tip to detect onset of binding between at least one target molecule and at least one affinity molecule;
c) interrogating the array of step b) with a probe having a tip to detect dissociation of at least one target molecule and at least one affinity molecule; and
d) measuring the time between onset of binding detected in step c) and dissociation detected in step c) to determine target occupancy time.
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Specification