Polymers and DNA copolymer coatings
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.
1. A substrate having a first surface comprising a polymer covalently bonded thereto, wherein the polymer comprises a recurring unit of Formula (IV):
wherein each of R 1c and R 2c is independently hydrogen, optionally substituted alkyl, or optionally substituted phenyl;
R 3c is hydrogen, optionally substituted alkyl, optionally substituted phenyl, or optionally substituted C 7-14 aralkyl;
Ar is an optionally substituted C 6-10 aryl or an optionally substituted 5 or 6 membered heteroaryl;
R A is optionally substituted tetrazine; and
L 3 is a single bond, an optionally substituted alkylene linker, or an optionally substituted heteroalkylene linker.
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):
wherein R 1c is selected from hydrogen or methyl.
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:
wherein each R 4a , R 4b and R 5b is independently hydrogen or C 1-3 alkyl; and
each 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 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
or combinations thereof, and wherein * indicates the polymer's point of connection with the first surface.
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:
wherein the covalent bonding position with the functionalized nucleotide is shown by a bond with a wavy line.
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):
wherein each of R 1c and R 2c is independently hydrogen, optionally substituted alkyl, or optionally substituted phenyl;
R 3c is hydrogen, optionally substituted alkyl, optionally substituted phenyl, or optionally substituted C 7-14 aralkyl;
Ar is an optionally substituted C 6-10 aryl or an optionally substituted 5 or 6 membered heteroaryl;
R A is optionally substituted tetrazine; and
L 3 is a single bond, an optionally substituted alkylene linker, or an optionally substituted heteroalkylene linker.
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;
wherein each of R 1c and R 2c is independently hydrogen, optionally substituted alkyl, or optionally substituted phenyl;
R 3c is hydrogen, optionally substituted alkyl, optionally substituted phenyl, or optionally substituted C 7-14 aralkyl;
Ar is an optionally substituted C 6-10 aryl or an optionally substituted 5 or 6 membered heteroaryl;
R A is optionally substituted tetrazine; and
L 3 is a single bond, an optionally substituted alkylene linker, or an optionally substituted heteroalkylene linker.
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.