Methods for generating ligand arrays

• US 6,919,181 B2
• Filed: 03/25/2002
• Issued: 07/19/2005
• Est. Priority Date: 03/25/2002
• Status: Expired due to Fees

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Hargreaves, John S.
• Primary Examiner(s): Forman, B J
• Application Number:
  US10/106,685
• Publication Number:
  US 20030180732A1
• Time in Patent Office: 1,212 Days
• Field of Search: 435/6, 435/7.1, 435/174, 435/283.1, 438/283.1, 438/287.2, 536/23.1, 536/25.3
• US Class Current: 435/6.11
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence

Abstract

Methods of producing ligand arrays, e.g., polypeptide and nucleic acid arrays, as well as the arrays produced thereby, methods for use of the arrays and kits that include the same, are provided. In the subject methods, a substrate having a surface displaying carboxylic ester functional groups that produce surface bound hydroxyl functional groups upon hydrolysis is first provided. Next, the functional groups are hydrolyzed to hydroxyl functional groups. The resultant substrate, optionally after an additional functionalization step, is then contacted with ligands, e.g., via deposition of each different ligand onto a different region of the surface, resulting in covalent attachment of the contacted ligands to the surface.

Claims

1. A method of producing an array of at least two different ligands covalently bonded to a surface of a substrate, said method comprising:

(a) providing a silica comprising substrate having a surface displaying carboxylic ester functional groups that produce surface bound hydroxyl functional groups and a free carboxylic acid upon hydrolysis, wherein said carboxylic ester functional groups are moieties of silane molecules covalently bonded to said surface;

(b) converting said carboxylic ester functional groups to produce a free carboxylic acid and a substrate having a surface displaying hydroxyl functional groups; and

(c) producing at least two different ligands covalently bound to said surface to produce said array by either;

contacting said surface with said at least two different ligands;

or

in situ synthesizing said ligands on said surface.

2. The method according to claim 1, wherein said covalently bound ligands are produced on said surface by contacting said surface with said at least two different ligands.

3. The method according to claim 1, wherein said covalently bound ligands are produced on said surface by in situ synthesis of said ligands on said surface.

4. The method according to claim 1, wherein said substrate is glass.

5. The method according to claim 1, wherein said converting step (b) comprises subjecting said surface to acid hydrolysis conditions.

6. The method according to claim 1, wherein said converting step (b) comprises subjecting said surface to alkali hydrolysis conditions.

7. The method according to claim 1, wherein said ligands are polymers.

8. The method according to claim 7, wherein said polymers are nucleic acids.

9. The method according to claim 7, wherein said polymers are peptides.

10. A method of producing an array of at least two different polymer ligands covalently attached to a surface of a substrate, said method comprising:

(a) providing a silica comprising substrate having a surface modified with silane molecules having carboxylic ester functional groups that produce surface bound hydroxyl functional groups and a free carboxylic acid upon hydrolysis, wherein said carboxylic ester functional groups are moieties of silane molecules covalently bonded to said surface;

(b) converting said carboxylic ester functional groups to produce a free carboxylic acid and a substrate having a surface displaying hydroxyl functional groups; and

(c) contacting said surface of said substrate displaying hydroxyl functional groups with said at least two different polymer ligands to covalently bond said at least two different polymer ligands to said surface and produce said array.

11. The method according to claim 10, wherein said substrate is glass.

12. The method according to claim 10, wherein said converting step (b) comprises subjecting said surface to acid hydrolysis conditions.

13. The method according to claim 10, wherein said converting step (b) comprises subjecting said surface to alkali hydrolysis conditions.

14. The method according to claim 10, wherein said polymer ligands are nucleic acids.

15. The method according to claim 10, wherein said polymer ligands are peptides.

16. A method of producing an array of at least two different nucleic acids covalently attached to a surface of a substrate, said method comprising:

(a) providing a silica comprising substrate having a surface displaying carboxylic ester functional groups that produce surface bound hydroxyl functional groups upon acid hydrolysis, wherein said carboxylic ester functional groups are moieties of silane molecules covalently bonded to said surface;

(b) converting said carboxylic ester functional groups to hydroxyl functional groups to produce a free carboxylic acid and a surface displaying hydroxyl functional groups; and

(c) depositing each of said least two different nucleic acids onto different regions of said surface to covalently bond said at least two different nucleic acids to said surface and produce said array.

17. The method according to claim 16, wherein said nucleic acids are oligonucleotides.

18. The method according to claim 16, wherein said nucleic acids are polynucleotides.

19. The method according to claim 18, wherein said polynucleotides are cDNAs.

20. A method of producing a surface modified substrate, said method comprising:

(a) providing a silica comprising substrate having a surface displaying carboxylic ester functional groups that produce surface bound hydroxyl functional groups and a free carboxylic acid upon acid hydrolysis, wherein said carboxylic ester functional groups are moieties of silane molecules covalently bonded to said surface;

(b) converting said carboxylic ester functional groups to surface bound hydroxyl functional groups.

21. The method according to claim 20, wherein said substrate is glass.

22. The method according to claim 20, wherein said converting step (b) comprises subjecting said surface to acid hydrolysis conditions.

23. The method according to claim 20, wherein said converting step (b) comprises subjecting said surface to alkali hydrolysis conditions.