Polymers and DNA copolymer coatings

Patent metadata

Field	Value
Publication number	US-10208142-B2
Application number	US-201715809656-A
Country	US
Kind code	B2
Filing date	Nov 10, 2017
Priority date	Oct 31, 2014
Publication date	Feb 19, 2019
Grant date	Feb 19, 2019

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.

First claim

What is claimed is:

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

wherein:
each of R 1a , R 2a , R 1b and R 2b is independently selected from the group consisting of hydrogen and optionally substituted alkyl;
each of R 3a and R 3b is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted phenyl, and optionally substituted C 7-14 aralkyl; and
each of L 1 and L 2 is independently selected from the group consisting of an optionally substituted alkylene linker and an optionally substituted heteroalkylene linker.

2. The substrate of claim 1 , wherein:
each of R 2a and R 3a is hydrogen;
each of R 2b and R 3b is hydrogen;
L 1 is an optionally substituted alkylene; or
L 2 is an optionally substituted alkylene.

3. The substrate of claim 1 , wherein Formula (I) is represented by Formula (Ia) and Formula (II) is represented by Formula (IIa):

wherein R 1a is hydrogen or methyl and R 1b is hydrogen or methyl.

4. The substrate of claim 1 , wherein the recurring unit of Formula (I) is represented by Formula (Ib):

wherein R 1a is hydrogen or methyl.

5. The substrate of claim 1 , further comprising 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 1 , further comprising one or more recurring units of Formula (IIIa) or (IIIb) or both:

wherein
each of R 4a , R 4b , and R 5b is independently hydrogen or C 1-3 alkyl; and
each of R 5a , R 6a , R 6b and R 7b is independently hydrogen, optionally substituted C 1-6 alkyl, or optionally substituted phenyl.

7. The substrate of claim 6 , wherein:
each of R 4a , R 4b , and R 5b is independently hydrogen or methyl;
R 6b and R 7b are each hydrogen;
at least one of R 5a or R 6a is hydrogen or methyl.

8. The substrate of claim 6 , wherein the one or more recurring units of Formula (IIIa) are represented by Formula (IIIa1), (IIIa2), or (IIIa3):

9. The substrate of claim 6 , wherein the one or more recurring units of Formula (IIIb) are represented by Formula (IIIb1):

10. The substrate of claim 1 , wherein the substrate surface comprises functionalized silane comprising unsaturated moieties selected from the group consisting of cycloalkenes, cycloalkynes, heterocycloalkenes, and heterocycloalkynes.

11. The substrate of claim 1 , further comprising functionalized oligonucleotides covalently bonded to the polymer.

12. The substrate of claim 11 , wherein the covalent bonding between the functionalized oligonucleotide and the polymer comprises the structural moiety:

or combinations thereof,
wherein * indicates the point of connection with the functionalized oligonucleotide.

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

wherein the covalent bonding position with the functionalized nucleotide is shown by a bond with a wavy line, and wherein is a single or double bond.

14. 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 functional oligonucleotides, which are selected from the group consisting of alkynes, cycloalkenes, cycloalkynes, heterocycloalkenes, and heterocycloalkynes, and optionally substituted variants thereof, and combinations thereof.

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

16. The substrate of claim 11 , wherein the covalent bond between the functionalized oligonucleotide and the polymer is the product of an azide click chemistry reaction.

17. The substrate of claim 16 , wherein the functionalized oligonucleotides are covalently bonded to the polymer through reaction of one or more functionalized moieties of the functionalized oligonucleotides with azido groups of the polymer, wherein the one or more functional moieties are selected from the group consisting of alkynes, cycloalkenes, cycloalkynes, heterocycloalkenes, and heterocycloalkynes, and optionally substituted variants thereof, and combinations thereof.

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

19. The substrate of claim 18 , wherein the polymer-coated regions are wells and the inert regions are interstitial regions.

20. A method of immobilizing a polymer to a first surface of a substrate comprising:
contacting a polymer comprising a recurring unit of Formula (I) and a recurring unit of Formula (II) 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;

wherein:
each of R 1a , R 2a , R 1b and R 2b is independently selected from the group consisting of hydrogen and optionally substituted alkyl;
each of R 3a and R 3b is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted phenyl, and optionally substituted C 7-14 aralkyl; and
each of L 1 and L 2 is independently selected from the group consisting of an optionally substituted alkylene linker and an optionally substituted heteroalkylene linker.

21. The method of claim 20 , wherein the first plurality of functional groups comprise vinyl, acryloyl, alkenyl, cycloalkenyl, heterocycloalkenyl, alkynyl, cycloalkynyl, heterocycloalkynyl, nitrene, aldehyde, hydrazinyl, glycidyl ether, epoxy, carbene, isocyanate, and maleimide, or optionally substituted variants or combinations thereof.

22. The method of claim 20 , wherein the first plurality of functional groups comprise epoxy or norbornene groups, or a combination thereof.

Assignees

• Illumina Cambridge Ltd
• Illumnia Cambridge Ltd

Inventors

• Brown Andrew A
• George Wayne N
• Richez Alexandre
• Dingwall Anne-Cecile
• Von Hatten Xavier

Classifications

• C08F220/282

  and containing two or more oxygen atoms · CPC title

• C08F220/346

  and further oxygen · CPC title

• C12Q1/6874

  involving nucleic acid arrays, e.g. sequencing by hybridisation · CPC title

• C08F20/60 Primary

  containing nitrogen in addition to the carbonamido nitrogen · CPC title

• B01J2219/00637

  by coating it with another layer · CPC title