Silane mixtures
View Patent ↗Silanation compositions containing a mixture of two or more silanation reagents, where at least one silanation reagent includes a functional group capable of supporting polymer synthesis and at least one silanation reagent includes no functional group capable of supporting polymer synthesis are useful in modulating the active site density and hydrolytic stability of a surface. These compositions are particularly useful in silanating a surface prior to preparation of a polymer array and provide for increased hybridization results.
1. A method of preparing an array of polymers comprising:
(a) contacting a silica or other silicon oxide-containing surface of a substrate with a silanation composition that comprises a mixture of two or more silanation reagents, each comprising a trialkoxy silyl group and wherein at least one silanation reagent includes a functional group capable of supporting polymer synthesis and at least one silanation reagent includes no functional group capable of supporting polymer synthesis and the molar ratio of the silanation reagent with no functional group to the silanation reagent with a functional group is from about 50:1 to 500:1 respectively, wherein the functional group capable of supporting polymer synthesis is protected with a protecting group or the functional group capable of supporting polymer synthesis is reacted with a compound having a reactive group protected by a protecting group;
(b) removing the protecting group in selected regions of the surface to provide an exposed functional or reactive group;
(c) reacting said exposed group with a monomer, wherein the monomer is coupled to the exposed group; and
(d) repeating the steps of removing and reacting to produce the array of polymers.
2. A method of preparing an array of polymers according to claim 1 wherein the polymer is a nucleic acid.
3. A method of preparing an array of polymers according to claim 1 , wherein the protecting group is a photoremovable protecting group.
4. A method of preparing an array of polymers according to claim 1 , wherein the protecting group is an acid labile protecting group.
5. A method of preparing an array of polymers according to claim 1 , wherein the step of removing comprises activating a photoacid generator in the selected regions by selective application of light having a predetermined wavelength to provide an acid and exposing the protecting group to the acid.
6. A method of preparing an array of polymers according to claim 1 wherein the at least one silanation reagent which includes a functional group capable of supporting polymer synthesis is selected from the group consisting of:
N-(3-(triethoxysilyl)propyl)-4-hydroxybutyramide
and wherein the at least one silane reagent which does not include a functional group capable of supporting polymer synthesis is selected from the group consisting of alkyl-trimethoxysilane, 1,2-bis(trimethoxysilyl)ethane, 1,3-bis(trimethoxysilyl)propane, 1,6-bis(trimethoxysilyl)hexane, 3-(N,N-dimethylamino)propyltrimethoxysilane and N,N-bis(3-trimethoxysilylpropyl)methylamine.
7. A method of preparing an array of polymers according to claim 1 wherein the at least one silanation reagent which includes a functional group capable of supporting polymer synthesis is N-(2-hydroxyethyl)-N,N-bis(trimethoxysilylpropyl)amine and the at least one silanation reagent which includes no functional group capable of supporting polymer synthesis is 1,2-bis(trimethoxysilyl)ethane.
8. A method of preparing an array of polymers according to claim 1 wherein the at least one silanation reagent which includes no functional group capable of supporting polymer synthesis and at the least one silanation reagent which includes a functional group capable of supporting polymer synthesis are present at predetermined ratios.
9. A method of preparing an array of polymers according to claim 1 wherein the molar ratio ranges from about 50 to 1 to 200 to 1, respectively.
10. A method of preparing an array of polymers according to claim 9 wherein the ratio ranges from about 50 to 1 to 100 to 1, respectively.
11. A method of preparing an array of polymers according to claim 10 wherein the ratio is about 99 to 1, respectively.
12. A method of preparing an array of polymers according to claim 1 wherein the at least one silanation reagent which includes no functional group capable of supporting polymer synthesis is 1,2-bis(trimethoxysilyl)ethane and the at least one silanation reagent which includes a functional group capable of supporting polymer synthesis is bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane.
13. A method of preparing an array of polymers according to claim 12 wherein the molar ratio of 1,2-bis(trimethoxysilyl)ethane to bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane ranges from about 50 to 1 to 500 to 1, respectively.
14. A method of preparing an array of polymers according to claim 13 wherein the molar ratio of 1,2-bis(trimethoxysilyl)ethane to bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane ranges from about 50 to 1 to 200 to 1, respectively.
15. A method of preparing an array of polymers according to claim 14 wherein the molar ratio of 1,2-bis(trimethoxysilyl)ethane to bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane ranges from about 50 to 1 to 100 to 1, respectively.
16. A method of preparing an array of polymers according to claim 15 wherein the molar ratio of 1,2-bis(trimethoxysilyl)ethane to bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane ratio is about 99 to 1, respectively.
17. A method of preparing an array of polymers according to claim 1 wherein the composition does not comprise a mixture of bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane and 1,2-bis(trimethoxysilyl)ethane.
18. A method of preparing an array of polymers according to claim 1 wherein the composition does not contain 1,2-bis(trimethoxysilyl)ethane.
19. The method of claim 1 , wherein the removing and reacting steps result in coupling of the monomer at an efficiency of 90% or more.