Method of making interactive protein arrays

• US 20030082560A1
• Filed: 10/29/2001
• Published: 05/01/2003
• Est. Priority Date: 10/29/2001
• Status: Abandoned Application

• Current Assignee: Yingjian Wang
• Original Assignee: Yingjian Wang
• Inventors: Wang, Yingjian
• Application Number:
  US10/055,838
• Publication Number:
  US 20030082560A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C40B 30/04 by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
• CPC Class Codes: G01N 33/6842 Proteomic analysis of subsets of protein mixtures with reduced complexity, e.g. membrane proteins, phosphoproteins, organelle proteins
• CPC Class Codes: G01N 33/6845 Methods of identifying protein-protein interactions in protein mixtures

Abstract

A device for biomedical research contains a flat solid support and a plurality of proteins, with the proteins immobilized on the support at pre-determined positions. The proteins immobilized at each position can be different but share a common property. An interactive protein array is produced when proteins that interact with a ligand are immobilized at a position and proteins interact with another ligand are immobilized at another position on the flat support. The interactive protein arrays are produced either by isolating interacting proteins sequentially or simultaneously. Chemical covalent cross-linking is used to stabilize the protein complexes on the array and to immobilize the proteins on the flat support. The devices are used in studying proteins, particularly in identifying protein-protein interactions and their modulators.

Claims

1. An array of biological reagents immobilized on a solid support, said array having separate known sites, said reagents at each said site interacting directly or indirectly with one or more ligands, and said reagents being covalently cross-linked to said ligands.

2. The array of claim 1, wherein said array has between 100 and 100,000 sites

3. The array of claim 1, wherein said ligands are antibodies.

4. The array of claim 1, wherein said ligands are recombinant proteins.

5. The array of claim 1, wherein said ligands are purified proteins.

6. The array of claim 1, wherein said ligands are nucleic acids.

7. The array of claim 1, wherein said ligands are oligo nucleotides.

8. A method of making an array of biological reagents, comprising the steps of

immobilizing a plurality of ligands on a solid support, each said ligand at a separate known site;

contacting said ligands on said solid support with a plurality of said reagents to allow the binding between said ligands and said reagents;

covalently cross-linking said ligands and said reagents.

9. The method of claim 8, wherein said ligands are antibodies.

10. The method of claim 8, wherein said ligands are recombinant proteins.

11. A method of making an array of biological reagents, comprising the steps of

immobilizing a plurality of ligands on a first solid support, each said ligand at a separate known site;

contacting said ligands on said solid support with a plurality of said reagents to allow the binding between said ligands and said reagents;

transferring and immobilizing said reagents bound to said ligands onto a second solid support.

12. The method of claim 11, wherein said ligands are covalently immobilized on said first solid support.

13. The method of claim 11, wherein said reagents are non-covalently immobilized on said second solid support.

14. The method of claim 11, wherein said ligands are antibodies.

15. The method of claim 11, wherein said ligands are recombinant proteins.

16. A method of making an array of biological reagents on a solid support, comprising the steps of

preparing said reagents by using ligands that bind said reagents;

immobilizing said reagents on said solid support.

17. The method of claim 16, wherein said ligands are antibodies.

18. The method of claim 16, wherein said ligands are recombinant proteins.

19. The method of claim 16, wherein said ligands are immobilized on membranes made of materials selected from the group consisting of nitrocellulose, nylon, polyvinylidene difluoride, glass, or plastics, and their derivatives.

20. A method of biological assays, comprising the steps of:

immobilizing a plurality of ligands on a solid support, each said ligand at a separate know site;

incubating said ligands on said solid support with a plurality of reagents to allow the binding between said ligands and said reagents;

covalently cross-linking said ligands and said reagents.

21. The method of claim 20, wherein one side of said support is at least 1 millimeter long.

22. The method of claim 20, wherein said ligands are antibodies.

23. The method of claim 20, wherein the number of said ligands ranges from 1 to 1,000,000.

24. The method of claim 20, wherein the number of said ligands ranges from 100 to 10,000.

25. A kit useful for detecting proteins, the kit containing a solid support, on which a plurality of reagents are immobilized, each at a predetermined position, said reagents at each said position interacting with one or a plurality of ligands.

26. The kit of claim 25, wherein said reagents are proteins.

27. The kit of claim 25, wherein said ligands are antibodies.

28. The kit of claim 25, wherein said ligands are recombinant proteins.

29. The kit of claim 25, wherein said reagents are covalently cross-linked with said ligands.