METHODS OF ANALYZING BINDING INTERACTIONS
The present disclosure relates to methods of analyzing binding interactions between a binding component and a receptor component by translocating unbound and any bound components through a pore and detecting the unbound and bound components.
1 . A method of analyzing binding interactions in solution, comprising:
(a) contacting one or more binding components with a receptor component, wherein unbound and any bound components are capable of translocating through a pore;
(b) detecting the unbound and any bound components by translocating the unbound and bound components through the pore, wherein the detecting is by imaging the charge induced field effect; and
(c) determining the number of binding component bound per number of receptor component.
2 . The method of claim 1 in which the pore comprises a nanopore.
3 . The method of claim 1 in which the unbound and any bound components are counted.
4 . The method of claim 1 in which the number of binding component bound per number of receptor component is determined at different concentrations of binding and receptor components.
5 . The method of claim 1 in which the number of binding component bound per number of receptor component is determined at different concentrations of binding component and at constant concentration of receptor component.
6 . The method of claim 1 in which the receptor component has n interaction sites with the binding component.
7 . The method of claim 6 for the binding interaction in which n is 1.
8 . The method of claim 6 for the binding interaction in which n is 2 to 10.
9 . The method of claim 6 for the binding interaction in which the interaction sites have the same equilibrium binding constants.
10 . The method of claim 6 for the binding interaction in which at least two of the interaction sites have different equilibrium binding constants.
11 . The method of claim 6 for the binding interaction in which at least two of the interaction sites display cooperative interactions.
12 . The method of claim 6 in which the interaction sites comprise at least a first and second interaction sites, wherein the first and second interactions sites are different.
13 . The method of claim 1 for the binding interaction in which the binding component and the receptor component are the same.
14 . The method of claim 1 for the binding interaction in which the binding component and receptor component are different.
15 . The method of claim 1 in which the binding components comprise at least a first and second binding components, wherein the first and second binding components are different.
16 . The method of claim 15 in which the first and second binding components bind the receptor component competitively.
17 . The method of claim 15 in which the first and second binding components bind the receptor component non-competitively.
18 . The method of claim 1 in which the binding component is a member of a library of binding components, and the binding interactions of each member of the library is analyzed.
19 . The method of claim 1 in which the receptor component comprises a protein that that binds the binding component.
20 . The method of claim 19 in which the protein comprises an enzyme.
21 . The method of claim 19 in which the protein comprises a cellular receptor.
22 . The method of claim 21 in which the cellular receptor comprises a cell-surface receptor.
23 . The method of claim 19 in which the binding component comprises an agonist of the protein.
24 . The method of claim 19 in which the binding components comprises an antagonist of the protein.
25 . The method of claim 19 in which the binding component comprises a small organic molecule of about 500 to about 3000 Daltons.
26 . The method of claim 19 in which the protein comprises an antibody.
27 . The method of claim 26 in which the binding component comprises an antigen bound by the antibody.
28 . The method of claim 1 in which the receptor component comprises an oligonucleotide.
29 . The method of claim 28 in which the binding component comprises a small organic molecule of about 500 to 3000 Daltons that binds the oligonucleotide.
30 . The method of claim 28 in which the binding component comprises a protein that binds the oligonucleotide.
31 . The method of claim 28 in which the binding component comprises a substantially complementary oligonucleotide.
32 . The method of claim 31 in which the substantially complementary oligonucleotide has at least a single nucleotide mismatch.