Polymer coated surfaces for microarray applications

• US 6,413,722 B1
• Filed: 03/22/2000
• Issued: 07/02/2002
• Est. Priority Date: 03/22/2000
• Status: Expired due to Fees

• Current Assignee: Incyte Pharmaceuticals, Inc. (Incyte Corporation)
• Original Assignee: Incyte Genomics, Inc. (Incyte Corporation)
• Inventors: Sawan, Samuel P., Arnold, Lyle J., Lee, Paul H.
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): FORMAN, BETTY J
• Application Number:
  US09/532,419
• Time in Patent Office: 832 Days
• Field of Search: 435/6, 435/287.2, 435/288.3, 435/287.9, 536/23.1, 422/68.1
• US Class Current: 435/6.12
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/0072 Organic compounds
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: C03C 17/30 with silicon-containing compounds
• CPC Class Codes: C03C 17/3405 with at least two coatings of organic materials C03C17/36, C03C17/42 take precedence
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/552 Glass or silica

Abstract

Methods are provide for modifying a solid support, such as a glass slide, by silylating with an agent having the formula H₂N—(CH₂)_n—SiX₃ where n is between 1 and 10, and X is independently chosen from OMe, OEt, Cl, Br, or I, then activating with a crosslinking reagent, followed by reacting with an amine-containing polymer. The support can optionally be reacted with a crosslinking reagent again. The support thus modified may be used to make arrays and microarrays where a plurality of targets are stably associated with the support and arranged in a defined manner.

Claims

1. A method of attaching target polynucleotides to a surface of a solid support, the method comprising the ordered steps of:

a) silylating the surface with an agent having the formula H₂N—

(CH₂)_n—

SiX₃ where n is between 1 and 10, and X is independently chosen from OMe, OEt, Cl, Br, or I;

b) activating the surface with a first crosslinking reagent;

c) reacting the surface with an amine-containing polymer to form a modified solid support surface; and

d) attaching non-activated polynucleotide targets to the modified solid support surface, wherein the polynucleotide targets are arranged in a defined manner on the modified solid support surface thereby producing an array of attached polynucleotides.

2. The method of claim 1, wherein after the reacting step, the solid support surface is further modified with a second crosslinking reagent, and wherein the polynucleotide targets are covalently attached to the modified solid support.

3. The method of claim 2, wherein the second crosslinking reagent is cyanuric chloride.

4. The method of claim 1, wherein the solid support is selected from the group consisting of glass, silica, plastic, ceramic, beads, and nylon, and combinations thereof.

5. The method of claim 1, wherein the solid support is glass.

6. The method of claim 1, wherein the crosslinking reagent is cyanuric chloride.

7. The method of claim 1, wherein the amine-containing polymer is selected from the group consisting of polyethylenimine, polyhistidine, polylysine, and polyarginine, and combinations thereof.

8. The method of claim 1, wherein the amine-containing polymer is polyethylenimine.

9. The method of claim 1, wherein the polynucleotide targets independently comprise from about 2 to about 50000 nucleotides in length.

10. The method of claim 1, wherein the polynucleotide targets independently comprise from about 50 to 65 nucleotides in length.

11. The method of claim 1, wherein the polynucleotide targets comprise a 5′

-end and a 3′

-end and a spacer arm at the 5′

-end or the 3′

-end.

12. The method of claim 1, wherein the polynucleotide targets comprise a 5′

-end and a 3′

-end and a spacer arm at the 5′

-end or the 3′

-end and the polynucleotide targets are covalently attached to the modified solid support surface through the spacer arm.

13. The method of claim 1, wherein the polynucleotide targets are non-covalently attached to the modified solid support surface.

14. An array of attached target polynucleotides, the array made by the method of claim 1.

15. The array of claim 14, wherein after the reacting step, the solid support surface is further modified with a second crosslinking reagent, and wherein the polynucleotide targets are covalently attached to the modified solid support.

16. The array of claim 15, wherein the second crosslinking reagent is cyanuric chloride.

17. The array of claim 14, wherein the amine-containing polymer is selected from the group consisting of polyethylenimine, polyhistidine, polylysine, and polyarginine, and combinations thereof.

18. The array of claim 14, wherein the amine-containing polymer is polyethylenimine.

19. The array of claim 14, wherein the polynucleotides independently comprise from about 50 to about 65 nucleotides.

20. The array of claim 14, wherein the solid support is selected from the group consisting of glass, silica, plastic, ceramic, beads, and nylon, and combinations thereof.

21. The array of claim 14, wherein the solid support is glass.

22. The array of claim 14, wherein the crosslinking reagent is cyanuric chloride.

23. The array of claim 14, wherein the polynucleotide targets comprise a 5′

-end and a 3′

-end and a spacer arm at the 5′

-end or the 3′

-end.

24. The array of claim 14, wherein the polynucleotide targets comprise a 5′

-end and a 3′

-end and a spacer arm at the 5′

-end or the 3′

-end and the polynucleotide targets are covalently attached to the modified solid support surface through the spacer arm.

25. The array of claim 14, wherein the polynucleotide targets are non-covalently attached to the modified solid support surface.