METHODS OF ANALYZING BINDING INTERACTIONS

• US 20080050752A1
• Filed: 06/14/2007
• Published: 02/28/2008
• Est. Priority Date: 06/30/2006
• Status: Abandoned Application

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: SUN, Hongye, Livak, Kenneth
• Application Number:
  US11/763,406
• Publication Number:
  US 20080050752A1
• Time in Patent Office: Days
• US Class Current: 435/7.2
• CPC Class Codes: G01N 33/48721 Investigating individual macromolecules, e.g. by translocation through nanopores Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds

Abstract

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.

Claims

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.