Polymers and DNA copolymer coatings

• US 10,577,439 B2
• Filed: 02/11/2019
• Issued: 03/03/2020
• Est. Priority Date: 10/31/2014
• Status: Active Grant

• Current Assignee: Illumina Cambridge Limited (Illumina Incorporated)
• Original Assignee: Illumina Cambridge Limited (Illumina Incorporated)
• Inventors: Brown, Andrew A., George, Wayne N., Richez, Alexandre, Dingwall, Anne-Cecile, von Hatten, Xavier
• Primary Examiner(s): Jones, Robert S
• Application Number:
  US16/272,923
• Publication Number:
  US 20190194363A1
• Time in Patent Office: 386 Days
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C08F 20/60 containing nitrogen in addition to the carbonamido nitrogen
• CPC Class Codes: C08F 220/22 Esters containing halogen
• CPC Class Codes: C08F 220/282 and containing two or more oxygen atoms
• CPC Class Codes: C08F 220/325 containing glycidyl radical, e.g. glycidyl (meth)acrylate
• CPC Class Codes: C08F 220/346 and further oxygen
• CPC Class Codes: C08F 220/56 Acrylamide; Methacrylamide
• CPC Class Codes: C08F 220/60 containing nitrogen in addition to the carbonamido nitrogen
• CPC Class Codes: C08F 220/603 and containing oxygen in addition to the carbonamido oxygen and nitrogen
• CPC Class Codes: C08F 222/385 Monomers containing two or more (meth)acrylamide groups, e.g. N,N'-methylenebisacrylamide
• CPC Class Codes: C08F 2438/03 Use of a di- or tri-thiocarbonylthio compound, e.g. di- or tri-thioester, di- or tri-thiocarbamate, or a xanthate as chain transfer agent, e.g . Reversible Addition Fragmentation chain Transfer [RAFT] or Macromolecular Design via Interchange of Xanthates [MADIX]
• CPC Class Codes: C08F 285/00 Macromolecular compounds obtained by polymerising monomers on to preformed graft polymers
• CPC Class Codes: C08F 293/005 using free radical "living" or "controlled" polymerisation, e.g. using a complexing agent
• CPC Class Codes: C08F 8/32 by reaction with amines
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2565/401 Signal amplification by chemical polymerisation
• CPC Class Codes: C40B 50/18 using a particular method of attachment to the solid support

Abstract

Some embodiments described herein relate to new polymer coatings for surface functionalization and new processes for grafting pre-grafted DNA-copolymers to surface(s) of substrates for use in DNA sequencing and other diagnostic applications.

Claims

1. A substrate having a first surface comprising a polymer covalently bonded thereto, wherein the polymer comprises a recurring unit of Formula (IV):

2. The substrate of claim 1, wherein R^1c is hydrogen or optionally substituted alkyl and each R^2c and R^3c is hydrogen.

3. The substrate of claim 1, wherein Ar is an optionally substituted phenyl.

4. The substrate of claim 1, wherein the recurring unit of Formula (IV) is also represented by Formula (IVa):

5. The substrate of claim 1, further comprises one or more recurring units selected from the group consisting of polyacrylamides, polyacrylates, polyurethanes, polysiloxanes, silicones, polyacroleins, polyphosphazenes, polyisocyanates, poly-ols, and polysaccharides, and combinations thereof.

6. The substrate of claim 5, further comprises one or more recurring units of Formula (IIIa) or (IIIb) or both:

7. The substrate of claim 6, wherein the recurring unit of Formula (IIIa) is also represented by (IIIa1), (IIIa2) or (IIIa3):

8. The substrate of claim 6, wherein the recurring unit of Formula (IIIb) is also represented by (IIIb1):

9. The substrate of claim 8, wherein the first surface of the substrate comprises functionalized silane comprising unsaturated moieties selected from the group consisting of cycloalkenes, cycloalkynes, heterocycloalkenes, and heterocycloalkynes, and combinations thererof.

10. The substrate of claim 9, wherein the covalent bonding between the polymer and the first surface of the substrate comprises the structure moiety

11. The substrate of claim 9, further comprising functionalized oligonucleotides covalently bonded to the polymer.

12. The substrate of claim 11, wherein the polymer comprises modified recurring units of the structure:

13. The substrate of claim 11, wherein the functionalized oligonucleotides are covalently bonded to the polymer through reaction of one or more functional moieties on the functionalized oligonucleotides with optionally substituted tetrazine, and wherein the functional moieties comprises alkynes, cycloalkenes, cycloalkynes, heterocycloalkenes, heterocycloalkynes, or optionally substituted variants or combinations thereof.

14. The substrate of claim 13, wherein the functional moieties comprise alkynes or optionally substituted variants thereof.

15. The substrate of claim 1, comprising both polymer coated regions and inert regions.

16. The substrate of claim 15, wherein the polymer-coated regions are wells and the inert regions are interstitial regions.

17. A polymer for surface functionalization, wherein the polymer comprises a recurring unit of Formula (IV):

18. A method for immobilizing a polymer to a first surface of a substrate, comprising:

contacting a polymer comprising a recurring unit of Formula (IV) with the first surface of the substrate, said first surface comprising a first plurality of functional groups covalently attached thereto; and

reacting the first plurality of functional groups of the first surface with the polymer, thereby covalently bonding the polymer to the first surface of the substrate;

19. The method of claim 18, wherein the first plurality of functional groups of the first surface comprise vinyl, acryloyl, alkenyl, cycloalkenyl, heterocycloalkenyl, alkynyl, cycloalkynyl, heterocycloalkynyl, nitrene, aldehyde, hydrazinyl, glycidyl ether, epoxy, carbene, isocyanate or maleimide, or optionally substituted variants, or combinations thereof.

20. The method of claim 18, wherein the first plurality of functional groups comprise norbornene groups.