Macromolecular arrays on polymeric brushes and methods for preparing the same

• US 6,994,964 B1
• Filed: 08/31/2000
• Issued: 02/07/2006
• Est. Priority Date: 09/01/1999
• Status: Expired due to Term

• Current Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Frank, Curtis W., McGall, Glenn, Chang, Ying Chih
• Primary Examiner(s): Wang, Andrew
• Assistant Examiner(s): SHIBUYA, MARK LANCE
• Application Number:
  US09/652,962
• Time in Patent Office: 1,986 Days
• Field of Search: 435/7.1, 435/4, 435/6, 435/DIG.19, 435/39, 435/49, 436524-531, 536/25.3, 427/264, 525/272
• US Class Current: 435/6.11
• 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: C08F 289/00 Macromolecular compounds obtained by polymerising monomers on to macromolecular compounds not provided for in groups C08F251/00 - C08F287/00
• CPC Class Codes: C08F 290/06 Polymers provided for in subclass C08G
• CPC Class Codes: C08F 4/02 Carriers therefor
• CPC Class Codes: C08L 51/08 grafted on to macromolecular compounds obtained otherwise than by reactions only involving unsaturated carbon-to-carbon bonds
• CPC Class Codes: C08L 51/10 grafted on to inorganic materials
• CPC Class Codes: C40B 40/00 Libraries per se, e.g. arrays, mixtures
• CPC Class Codes: G01N 33/544 the carrier being organic

Abstract

Polymeric brush substrates and methods for their preparation are provided. Methods are also provided for preparing macromolecular arrays on such polymeric brush substrates. Using polymeric brush substrates allows control over functional site density as well as wettability and porosity of the substrate. These polymeric brushes are useful in solid-phase synthesis of arrays of peptides, polynucleotides or small organic molecules.

Claims

1. A method of preparing a polymeric brush substrate having a plurality of polynucleotides thereon for use in a polynucleotide array, the method comprising:

(a) providing a substrate to which one or more free radical initiators are covalently attached, wherein each free radical initiator has a radical generation site distal to the substrate;

(b) contacting the covalently attached substrate with monomers under conditions that promote free radical polymerization from the radical generation sites of the initiators to form a polymeric brush; and

(c) covalently attaching a plurality of polynucleotides to a plurality of reactive groups on the polymeric brush.

2. The method of claim 1, wherein step (b) comprises living free radical polymerization.

3. The method of claim 1, wherein the substrate comprises glass or silica.

4. The method of claim 1, wherein the monomers comprise a vinyl group.

5. The method of claim 1, wherein the polymer brush formed on the support comprises hydroxyl, amino, carboxyl, or sulfydryl groups or a combination thereof.

6. The method of claim 1, wherein the monomers comprise vinyl acetate.