Catalyst-free surface functionalization and polymer grafting
Some embodiments described herein relate to a substrate with a surface comprising a silane or a silane derivative covalently attached to optionally substituted cycloalkene or optionally substituted heterocycloalkene for direct conjugation with a functionalized molecule of interest, such as a polymer, a hydrogel, an amino acid, a nucleoside, a nucleotide, a peptide, a polynucleotide, or a protein. In some embodiments, the silane or silane derivative contains optionally substituted norbornene or norbornene derivatives. Method for preparing a functionalized surface and the use in DNA sequencing and other diagnostic applications are also disclosed.
1. A substrate comprising a first surface comprising silane or a silane derivative covalently bound, optionally through a linker, to a functionalized molecule through reaction of the functionalized molecule with a first plurality of unsaturated moieties selected from the group consisting of norbornene, heteronorbornenes, norbornene derivatives, and optionally substituted variants and combinations thereof covalently attached to said silane or silane derivative, wherein the functionalized molecule comprises poly(N-(5-azidoacetamidylpentyl)acrylamide-co-acrylamide) (PAZAM) and
oligonucleotides covalently attached to the functionalized molecule through a second plurality of unsaturated moieties on the oligonucleotides, the second plurality of unsaturated moieties are selected from the group consisting of cycloalkenes, cycloalkynes, heterocycloalkenes, heterocycloalkynes, alkynes, and optionally substituted variants and combinations thereof.
2. The substrate of claim 1 , wherein the linkers are present, and wherein the linkers are covalently attached between silicon atoms of said silane or silane derivative and the first plurality of unsaturated moieties.
3. The substrate of claim 2 , wherein the linkers are selected from the group consisting of optionally substituted alkylenes, optionally substituted heteroalkylenes, optionally substituted cycloalkylenes, optionally substituted heterocyclylenes, optionally substituted arylenes, optionally substituted heteroarylenes, optionally substituted polyethylene glycols, cleavable linkers, and combinations thereof.
4. The substrate of claim 3 , wherein the linkers are selected from the group consisting of optionally substituted alkylenes and optionally substituted heteroalkylenes.
5. The substrate of claim 1 , wherein said second plurality of unsaturated moieties is an optionally substituted bicyclo[6.1.0]non-4-yne.
6. The substrate of claim 1 , wherein said second plurality of unsaturated moieties is an optionally substituted alkyne.
7. The substrate of claim 1 , wherein the substrate is selected from the group consisting of a glass substrate, a silica substrate, a plastic substrate, a quartz substrate, a metal substrate, a metal oxide substrate, and combinations thereof.
8. The substrate of claim 1 , wherein the first surface is patterned with nanowells.
9. The substrate of claim 1 , wherein the first plurality of unsaturated moieties of said silane or silane derivative is optionally substituted norbornene.
10. The substrate of claim 9 , wherein the linkers are present, and wherein the linkers are covalently attached between silicon atoms of said silane or silane derivative and the optionally substituted norbornene, and wherein the linkers are optionally substituted alkylenes.
11. The substrate of claim 9 , wherein the second plurality of unsaturated moieties of said oligonucleotides is alkyne.
12. The substrate of claim 9 , wherein the second plurality of unsaturated moieties of said oligonucleotides is optionally substituted bicyclo[6.1.0]non-4-yne.