阅读说明：本技术 用于制造生物分子阵列的可旋涂的胺反应性多用途表面涂层的制备 (Preparation of spin-coatable amine-reactive multi-purpose surface coatings for fabrication of biomolecule arrays ) 是由 格伦·麦克加尔 于 2018-03-06 设计创作，主要内容包括：本发明涉及使用聚甲基倍半硅氧烷薄膜的共价结合薄膜衍生化基底表面的方法,其可作为生物分子阵列合成的平台。这些方法还可用于制备生物分子阵列原位固相合成的基底表面。(The present invention relates to a method of derivatizing a substrate surface with a covalently bonded film of polymethylsilsesquioxane film, which can serve as a platform for biomolecule array synthesis. These methods can also be used to prepare substrate surfaces for in situ solid phase synthesis of biomolecule arrays.)

1. A solid support comprising a poly (methylsilsesquioxane) -bound polymer comprising a compound of formula I:

wherein the linker is-L¹-L²-L³-；

L¹And L³At each occurrence, is independently C₁-C₆Alkylene or C₂-C₆Alkyleneoxy (alkoxylene) wherein said C is₁-C₆Alkylene and C₂-C₆The alkyleneoxy group may be unsubstituted or substituted with 1 to 3 groups selected from the group consisting of: c₁-C₃Alkyl radical, C₁-C₃Alkoxy, halide, cyanide and-N (R20) -;

L²is an arylene group;

R¹independently selected from the group consisting of:

provided that there is at least one R¹Is that

T¹May be free of radicals, hydrogen, C₁-C₆Alkyl, or initiator residue;

x is independently protected, or unprotected hydroxy, protected, or unprotected-NHR²¹Protected, or unprotected, -SH, protected, or protected-CO₂H, protected, or protected-CHO, protected, or unprotected-ONH₂Protected, or unprotected-NHNH₂，-N₃，-C≡CR²²Or a halide;

R²independently is H, -CH₃or-CH₂OCH₃；

R²⁰，R²¹And R²²Are each independently C₁-C₃An alkyl group;

the Capture Probe (Capture Probe) comprises at least one molecule selected from the group consisting of: peptides, proteins, glycosylated proteins, glycoconjugates, aptamers, sugars, polynucleotides, oligonucleotides and polypeptides;

p is an integer between 2 and 200;

a is an integer between 1 and 5;

b is an integer between 0 and 10;

c is an integer between 1 and 5; and is

d is an integer between 0 and 10.

2. The solid support of claim 1, wherein said polymethylsilsesquioxane comprises a plurality of formulas (CH)₃SiO_3/2) A group of said plurality of formulae (CH) covalently bound to a linker of a compound of formula I₃SiO_3/2) The repeating unit of (1).

3. The solid support of claim 1, wherein the capture probe is an oligonucleotide.

4. The solid support of claim 3, wherein the capture probe is DNA.

5. The solid support of claim 1, further comprising a glass substrate, silica, silicon, fused silica substrate, metal or polymer substrate having the polymethylsilsesquioxane bound thereto.

6. The solid support of claim 5, wherein the substrate is a fused silica substrate.

7. The solid support of claim 5, wherein said polymeric substrate comprises at least one member selected from the group consisting of: acrylonitrile-butadiene-styrene, cyclic olefin polymers, cyclic olefin copolymers, polymethyl methacrylate, polycarbonate, polystyrene (Polystyrole), polypropylene, polyvinyl chloride, polyamide, polyethylene terephthalate, polytetrafluoroethylene, polyoxymethylene, thermoplastic elastomers, thermoplastic polyurethanes, polyimide, polyetheretherketone, polylactic acid, polymethylpentene.

8. The solid support of claim 2, wherein said plurality of formulas (CH)₃SiO_3/2) And the set of pluralities (CH)₃SiO_3/2) The ratio of repeating units is from about 15 to about 27.

9. The solid support of claim 8, wherein the ratio is from about 18 to about 24.

10. The solid support of claim 8, wherein the ratio is from about 20 to about 22.

11. A method of derivatizing a surface of a substrate comprising:

(a) contacting a surface of a substrate with a first agent comprising a polymethylsilsesquioxane-bound polymer comprising a compound of formula I

Wherein the linker is-L¹-L²-L³-；

L¹And L³At each occurrence, is independently C₁-C₆Alkylene, or C₂-C₆Alkyleneoxy (alkoxylene) wherein said C is₁-C₆Alkylene and C₂-C₆The alkyleneoxy group may be unsubstituted or substituted with 1 to 3 groups selected from the group consisting of: c₁-C₃Alkyl radical, C₁-C₃Alkoxy, halide, cyanide and-N (R)²⁰)-；

L²Is an arylene group;

R¹is a pentafluorophenoxy group;

R²⁰is C₁-C₃An alkyl group;

T¹may be free of radicals, hydrogen, C₁-C₆Alkyl, or initiator residue; and

p is an integer between 2 and 200; and

(b) making a first group R in the polymer¹With a second reagent comprising:

wherein the Capture Probe (Capture Probe) comprises at least one molecule selected from the group consisting of: peptides, proteins, glycosylated proteins, glycoconjugates, aptamers, sugars, polynucleotides, oligonucleotides and polypeptides;

a is an integer between 1 and 5; and is

b is an integer between 0 and 10.

12. The method of claim 11, wherein the polymethylsilsesquioxane comprises a plurality of formulas (CH)₃SiO_3/2) And a set of a plurality of formulae (CH) covalently bound to a linker of the compound of formula I₃SiO_3/2) The repeating unit of (1).

13. The method of claim 11, further comprising, prior to (a), drying or washing the surface of the substrate, or treating the surface to provide a plurality of hydroxyl groups.

14. The method of claim 11, wherein said surface of said substrate comprises a plurality of hydroxyl groups and, after (a), a set of said plurality of hydroxyl groups is covalently bonded to said polymethylsilsesquioxane.

15. The method of claim 11, the contacting in (a) being a coating or spin coating.

16. The method of claim 11, further comprising annealing the substrate with the first reagent prior to (b).

17. The method of claim 16, wherein said annealing is performed at about 130 ℃ for about 1 to about 3 hours.

18. The method of claim 11, further comprising, after step (b), allowing a second group R in said polymer to exist¹Reacting with a third reagent selected from the group consisting of:

wherein X is independently a protected, or unprotected, hydroxy group, is a protected, or unprotected-NHR²¹Is protected, or unprotected-SH, is protected, or unprotected-CO₂H, is protected, or unprotected-CHO, is protected, or unprotected-ONH₂Is protected, or unprotected-NHNH₂，-N₃，-C≡CR²²Or a halide;

R²independently is H, -CH₃or-CH₂OCH₃；

R²¹And R²²Independently is C₁-C₃An alkyl group;

a is an integer between 1 and 5;

b is an integer between 0 and 10;

c is an integer between 1 and 5; and is

d is an integer between 0 and 10.

19. The method of claim 11, wherein the substrate is a glass, silica, silicon, fused silica substrate, metal, or polymer substrate comprising at least one selected from the group consisting of: acrylonitrile-butadiene-styrene, cyclic olefin polymers, cyclic olefin copolymers, polymethyl methacrylate, polycarbonate, polystyrene (Polystyrole), polypropylene, polyvinyl chloride, polyamide, polyethylene terephthalate, polytetrafluoroethylene, polyoxymethylene, thermoplastic elastomers, thermoplastic polyurethanes, polyimide, polyetheretherketone, polylactic acid, polymethylpentene.

20. The method of claim 19, wherein the substrate is a fused silica substrate.

21. The method of claim 19, wherein the substrate is a cyclic olefin copolymer or cyclic olefin multimer.

22. The method of claim 11 wherein all of said formula (CH)₃SiO_3/2) And a linker covalently bound to a compound of formula I₃SiO_3/2) Is from about 15 to about 27.

23. The method of claim 11, wherein the capture molecule comprises a first oligonucleotide.

24. The method of claim 23, further comprising, after step (b), confirming immobilization of the first oligonucleotide on the surface by detecting hybridization between the first oligonucleotide and a second oligonucleotide comprising a label and having a sequence complementary to the first oligonucleotide.

25. The method of claim 23, wherein the label is a fluorophore.