Global Proteomic Screening Of Random Bead Arrays Using Mass Spectrometry Imaging

• US 20120077688A1
• Filed: 06/29/2011
• Published: 03/29/2012
• Est. Priority Date: 06/30/2010
• Status: Active Grant

• Current Assignee: Ambergen Inc.
• Original Assignee: Ambergen Inc.
• Inventors: Bergo, Vladislav B., Lim, Mark J., Rothschild, Kenneth J.
• Granted Patent: US 9,523,680 B2
• Time in Patent Office: Days
• US Class Current: 506/9
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/6845 Methods of identifying protein-protein interactions in protein mixtures
• CPC Class Codes: G01N 33/6848 Methods of protein analysis involving mass spectrometry

Abstract

Methods for proteomic screening on random protein-bead arrays by mass spec is described. Photocleavable mass tags are utilized to code a protein library (bait molecules) displayed on beads randomly arrayed in an array substrate. A library of probes (prey) can be mixed with the protein-bead array to query the array. Because mass spec can detect multiple mass tags, it is possible to rapidly identify all of the interactions resulting from this mixing.

Claims

1. A method of detecting the interaction of prey molecules with bait molecules on an array of beads, comprising a) providing a mixture comprising first and second beads, said first bead comprising a first mass tag and a first bait molecule, said second bead comprising a second mass tag and a second bait molecule, wherein said first and second bait molecules and said first and second mass tags are different;

b) contacting said first and second bait molecules with a solution comprising a prey molecule, wherein said prey molecule comprises a third mass tag which is different than first and second mass tags;

c) making an array with said beads; and

d) subjecting said array to mass spec analysis under conditions wherein binding of said prey molecule to said bait molecule is detected by measuring mass tags.

2. The method of claim 1, wherein said mass tags are selected from the group consisting of polypeptides, nucleic acids, linear polymers, branched polymers.

3. The method of claim 2, wherein said mass tags are released by light and detected during the mass spectrometric analysis of step d).

4. The method of claim 1, wherein said first and second mass tag are coupled to bead through a binding agent.

5. The method of claim 4, wherein said binding agent is streptavidin.

6. The method of claim 1, wherein said prey molecule is an antibody.

7. The method of claim 1, wherein said prey molecule is a protein.

8. The method of claim 1, wherein said prey molecule is a polypeptide.

9. The method of claim 1, wherein said prey molecule is a nucleic acid.

10. The method of claim 1, wherein said prey molecule is a small drug compound.

14. The method of claim 1, wherein said mass spec analysis in step d) comprises measuring mass tags from individual beads.

15. The method of claim 1, wherein said mass spec analysis in step d) comprises MALDI-MS imaging of individual beads in said array.

16. The method of claim 1, wherein said array of step c) is imaged fluorescently prior to said mass spec analysis of step d).

17. The method of claim 1, wherein said array of step c) is imaged fluorescently subsequent to said mass spec analysis of step d).

18. The method of claim 1, wherein said array of step c) is a random array.

35. The method of claim 1, wherein said array of step c) is imaged fluorescently subsequent to said mass spec analysis of step d).

11. The method of claim one wherein third mass tag is coupled to prey molecule through a binding agent.

12. The method of claim 11 wherein binding agent is an antibody.

13. The method of claim 11, wherein said antibody is fluorescently labeled.

19. A method of detecting the interaction of prey molecules with bait molecules , comprising a) providing a mixture comprising first and second beads, said first bead comprising a first mass tag and a first bait molecule, said second bead comprising a second mass tag and a second bait molecule, wherein said first and second bait molecules and said first and second mass tags are different;

b) making an array with said beads;

c) contacting said first and second bait molecules with a solution comprising a prey molecule, wherein said prey molecule comprises a mass tag; and

d) subjecting said array to MALDI mass spec analysis under conditions wherein binding of said prey molecules to a bait molecule is detected.

20. The method of claim 19, wherein said mass tags are selected from the group consisting of polypeptides, nucleic acids, linear polymers, branched polymers.

21. The method of claim 20, wherein said mass tags are released by light and detected during the mass spectrometric analysis of step d).

22. The method of claim 19, wherein said first and second mass tag are coupled to bead through a binding agent.

23. The method of claim 22, wherein said binding agent is streptavidin.

24. The method of claim 19, wherein said prey molecule is an antibody.

25. The method of claim 19, wherein said prey molecule is a protein.

26. The method of claim 19, wherein said prey molecule is a polypeptide.

27. The method of claim 19, wherein said prey molecule is a nucleic acid.

28. The method of claim 19, wherein said prey molecule is a small drug compound.

32. The method of claim 19, wherein said mass spec analysis in step d) comprises measuring mass tags from individual beads.

33. The method of claim 19, wherein said mass spec analysis in step d) comprises MALDI-MS imaging of individual beads in said array.

34. The method of claim 19, wherein said array of step c) is imaged fluorescently prior to said mass spec analysis of step d).

36. The method of claim 19, wherein said array of step c) is a random array.

29. The method of claim one wherein third mass tag is coupled to prey molecule through a binding agent.

30. The method of claim 29 wherein binding agent is an antibody.

31. The method of claim 29, wherein said antibody is fluorescently labeled.