Methods of making microarrays with substrate surfaces having covalently bound polyelectrolyte films

• US 6,989,267 B2
• Filed: 07/02/2001
• Issued: 01/24/2006
• Est. Priority Date: 07/02/2001
• Status: Expired due to Fees

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Lefkowitz, Steven M., Kim, Namyong, Chan, Athene W. C.
• Primary Examiner(s): Sisson, Bradley L.
• Application Number:
  US09/900,760
• Publication Number:
  US 20030008413A1
• Time in Patent Office: 1,667 Days
• Field of Search: 435/6, 435/7, 435/287.1, 435/287.2, 536/23.1, 536/24.31, 536/24.33, 530/350, 530/387.1
• US Class Current: 435/287.2
• CPC Class Codes: B01J 2219/00274 Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

Abstract

Methods for stably associating a polyelectrolyte coating or film to a substrate surface, as well as the coated substrates produced thereby, are disclosed herein. In the subject methods, substrate surfaces are coated with a polyelectrolyte, and the coated substrate surfaces are treated with a bifunctional molecule under conditions sufficient to stably associate the film to the array surface. The subject methods find use in a variety of different applications, including the production of mircoarrarys.

Claims

1. A method for fabricating a microarray, said method comprising:

(a) producing a polyelectrolyte layer on at least one surface of a substrate;

(b) depositing a plurality of spots of ligands on said polyelectrolyte layer, said ligand spots arranged in an array pattern;

(c) contacting said polyelectrolyte layer with a reagent comprising;

(i) a passivating moiety that reacts with said polyelectrolyte layer, and

(ii) a substrate reactive functionality; and

,

(d) producing a covalent bond between said substrate reactive functionality and said substrate wherein said substrate reactive functionality is a trialkoxysilyl anhydride.

2. The method of claim 1, further comprising cross-linking said ligand spots onto said polyelectrolyte layer.

3. The method of claim 2, wherein said ligands comprise nucleic acids.

4. The method of claim 1, wherein said depositing step (b) comprises depositing a plurality of different aqueous volumes of ligands on said polyelectrolyte layer.

5. The method according to claim 4, wherein said ligands are nucleic acids.

6. The method according to claim 4, wherein said ligands are peptides.

7. The method according to claim 4, wherein said plurality of aqueous volumes are deposited by ink-jet deposition.

8. A method for fabricating a microarray, said method comprising:

(a) producing a polyamine layer on at least one silica surface of a substrate;

(b) depositing a plurality of spots of ligands on said polymine layer, said ligand spots arranged in an array pattern;

(c) contacting said polyamine layer with a solution of a trialkoxysilyl anhydride; and

(d) forming a covalent bond between said anhydride and said polyamine layer and a covalent silicon-oxygen bond between said trialkoxysilyl propyl anhydride and said silica surface on said substrate.

9. The method of claim 8, wherein said polyamine comprises poly(L-lysine).

10. The method of claim 8, wherein said trialkoxysilyl anhydride comprises a compound having the structure;

wherein n is either zero or an integer from 1 to 10, m is either zero or an integer from 1 to 10, and R is an alkyl or functionally terminated alkenyl group.

11. The method of claim 10, wherein n is an integer equal to 1 or 2, and m is either zero or an integer from 1 to 4.

12. The method of claim 8, wherein said trialkoxysilyl anhydride comprises 3-(triethoxysilyl)propyl-succinic anhydride.

13. The method of claim 8, further comprising cross-linking said ligand spots onto said polyamine layer.

14. The method of claim 8, wherein said ligands comprise nucleic acids.

15. The method of claim 8, wherein said depositing step (b) comprises depositing a plurality of different aqueous volumes of ligands on said polyelectrolyte layer.

16. The method according to claim 15, wherein said ligands are nucleic acids.

17. The method according to claim 15, wherein said ligands are peptides.

18. The method according to claim 15, wherein said plurality of aqueous volumes are deposited by ink-jet deposition.

19. A method for fabricating a microarray, said method comprising:

(a) producing a polyelectrolyte layer on at least one surface of a substrate;

(b) depositing a plurality of different aqueous volumes on said polyelectrolyte layer in an array pattern to produce an array of deposited spots, wherein each aqueous volume of said plurality comprises a polymer;

(c) contacting said polyelectrolyte layer with a reagent comprising;

(i) a passivating moiety that reads with said polyelectrolyte layer, and

(ii) a substrate reactive functionality; and

(d) producing a covalent bond between said substrate reactive functionality and said substrate, wherein said substrate reactive functionality is a trialkoxysilyl anhydride.

20. The method of claim 19, further comprising cross-linking polymers of said deposited spots onto said polyelectrolyte layer.

21. The method of claim 19, wherein said polymers are nucleic acids.

22. The method of claim 19, wherein said polymers are peptides.

23. The method according to claim 19, wherein said plurality of aqueous volumes are deposited by ink-jet deposition.

24. The method according to claim 19, wherein said polyelectrolyte is a polyamine.

25. The method of claim 24, wherein said polyamine comprises poly(L-lysine).

26. The method of claim 19, wherein said reagent is a trialkoxysilyl anhydride.

27. The method of claim 26, wherein said trialkoxysilyl anhydride comprises a compound having the structure:

wherein n is either zero or an integer from 1 to 10, m is either zero or an integer from 1 to 10, and R is an alkyl or functionally terminated alkenyl group.

28. The method of claim 27, wherein n is an integer equal a 1 or 2, and m is either zero or an integer from 1 to 4.

29. The method of claim 26, wherein said trialkoxysilyl anhydride comprises 3-(triethoxysilyl)propyl-succinic anhydride.

30. A method for fabricating a nucleic acid microarray, said method comprising:

(a) producing a polyamine layer on at least one silica surface of a substrate;

(b) depositing a plurality of different aqueous volumes of nucleic acids on said polyamine layer in an array pattern to produce an array of deposited nucleic acid spots;

(c) contacting said polyamine layer with a solution of a trialkoxysilyl anhydride; and

(d) forming a covalent bond between said anhydride an said polyamine layer and a covalent silicon-oxygen bond between said trialkoxysilyl propyl anhydride and said silica surface on said substrate.

31. The method of claim 30, wherein said polyamine comprises poly(L-lysine).

32. The method of claim 30, wherein said trialkoxysilyl anhydride comprises a compound having the structure:

wherein n is either zero or an integer from 1 to 10, m is either zero or an integer from 1 to 10, and R is an alkyl or functionally terminated alkenyl group.

33. The method of claim 32, wherein n is an integer equal 1 or 2, and m is either zero or an integer from 1 to 4.

34. The method of claim 30, wherein said trialkoxysilyl anhydride comprises 3-(triethoxysilyl)proply-succinic anhydride.

35. The method of claim 30, further comprising cross-linking said nucleic acid spots onto said polyamine layer.

36. The method of claim 35, wherein said trialkoxysilyl anhydride comprises a compound having the structure:

wherein n is either zero or an integer from 1 to 10, m is either zero or an integer from 1 to 10, and R is an alkyl or functionally terminated alkenyl group.

37. The method of claim 36, wherein n is an integer equal to 1 or 2, and m is either zero or an integer from 1 to 4.

38. A method for fabricating a peptide microarray, said method comprising:

(a) producing a polyamine layer on at least one silica surface of a substrate;

(b) depositing a plurality of different aqueous volumes of peptides on said polyamine layer in an array pattern to produce an array of deposited peptide spots;

(c) contacting said polyamine layer with a solution of a trialkoxysilyl anhydride; and

(d) forming a covalent bond between said anhydride an said polyamine layer and a covalent silicon-oxygen bond between said trialkoxysilyl propyl anhydride and said silica surface on said substrate.

39. The method of claim 38, wherein said polyamine comprises poly(L-lysine).

40. The method of claim 38, wherein said trialkoxysilyl anhydride comprises 3-(triethoxysilyl)propyl-succinic anhydride.

41. The method of claim 38, further comprising cross-linking said nucleic acid spots onto said polyamine layer.