Method for attachment of silylated molecules to glass surfaces

Abstract

A method for the efficient immobilization of silylated molecules such as silylated oligonucleotides or proteins onto unmodified surfaces such as a glass surface is provided. Also provided are compounds, devices, and kits for modifying surfaces such as glass surfaces.

1 - 56 . (canceled)  
     
     
         57 . A device comprising a surface with an immobilized molecule, wherein said surface is prepared by the method comprising the steps of: 
 (a) contacting Si(NCY) 4  wherein Y represents oxygen or sulfur with an agent so as to form a first reactive intermediate, said agent having a formula ii:     (R 1 )(R 2 )(R 3 )Si—X-Z  ii   wherein R 1 , R 2  and R 3  independently represents C 1 -C 6  alkoxy, C 1 -C 6  alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy; X represents linear or branched C 1 -C 20  alkyl or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; and Z represents a hydroxy or amino group, with the proviso that at least one of R 1 , R 2  or R 3  represents C 1 -C 6  alkoxy;    (b) contacting the first reactive intermediate with a molecule so as to form a second reactive intermediate;    (c) contacting the second reactive intermediate with said surface so as to immobilized the molecule onto said surface.    
     
     
         58 . The device of  claim 57  wherein the surface is a glass surface.  
     
     
         59 . The device of  claim 57 , wherein the molecule is a probe.  
     
     
         60 . The device of  claim 59 , wherein the probe comprises a protein, a peptide, a nucleic acid, a peptide nucleic acid, a linked nucleic acid, an amino acid, a nucleoside triphosphate, a carbohydrate, a lipid, a lipid bound protein, an aptamer, a virus, a cell fragment, or a whole cell.  
     
     
         61 . The device of  claim 60 , wherein the lipid bound protein comprises a G-protein coupled receptor.  
     
     
         62 . The device of  claim 59 , wherein the probe comprises an antibody, an antigen, a receptor, or a ligand.  
     
     
         63 . The device of  claim 62 , wherein the probe has been derivatized to contain one or more amino and/or hydroxyl groups.  
     
     
         64 - 90 . (canceled)  
     
     
         91 . The device of  claim 57  wherein the surface is a glass surface.  
     
     
         92 . The device of  claim 57  wherein the surface has at least one group that reacts with the reactive intermediate.  
     
     
         93 . The device of  claim 92  wherein the group comprises a hydroxyl or amino group.  
     
     
         94 . The device of  claim 57  wherein the agent is 4-aminophenyl trimethoxysilane, 4-aminophenyl triethoxysilane, 3-aminophenyltrimethoxysilane, 3-amenophenyltriethoxysilane, or mixtures thereof.  
     
     
         95 . The device of  claim 94  wherein the agent and tetraisocyanatosilane is present at a 4:1 molar ratio.  
     
     
         96 . The device of  claim 57  wherein the molecule and the first reactive intermediate is present in a 3:1 molar ratio.  
     
     
         97 . The device of  claim 57  wherein the first reactive intermediate has a formula iv: 
         (R 1 )(R 2 )(R 3 )Si—X-Z-CYNH—Si (NCY) 3   iv 
       wherein R 1 , R 2  and R 3  independently represents C 1 -C 6  alkoxy, C 1 -C 6  alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy; X represents linear or branched C 1 -C 20  alkyl or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; and Z represents oxygen or NH, with the proviso that at least one of R 1 , R 2 , or R 3  represents C 1 -C 6  alkoxy.  
     
     
         98 . The device of  claim 97  wherein R 1 , R 2  and R 3  represents methoxy, X represents phenyl, Y represents oxygen, and Z represents NH.  
     
     
         99 . The device of  claim 97  wherein the second reactive intermediate has a formula v: 
         (R 1 )(R 2 )(R 3 )Si—X-Z-CYNH—Si (NHCYL-M) 3   v 
       wherein R 1 , R 2  and R 3  independently represents C 1 -C 6  alkoxy, C 1 -C 6  alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy; L represents a linking group; X represents linear or branched C 1 -C 20  alkyl or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; and Z represents oxygen or NH; and M represents a molecule, with the proviso that at least one of R 1 , R 2 , or R 3  represents C 1 -C 6  alkoxy.  
     
     
         100 . The device of  claim 99  wherein L represents a nucleophilic group from the molecule.  
     
     
         101 . The device of  claim 100  wherein the nucleophilic group comprises —NH, —S—, —O—, or —OOC—.  
     
     
         102 . The device of  claim 57  wherein the first reactive intermediate has a formula vi: 
         [(R 1 )(R 2 )(R 3 )Si—X-Z-CYNH] 2 —Si (NCY) 2   vi 
       wherein R 1 , R 2  and R 3  independently represents C 1 -C 6  alkoxy, C 1 -C 6  alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy; X represents linear or branched C 1 -C 20  alkyl or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; and Z represents oxygen or NH, with the proviso that at least one of R 1 , R 2 , or R 3  represents C 1 -C 6  alkoxy.  
     
     
         103 . The device of  claim 102  wherein R 1 , R 2  and R 3  represents methoxy, X represents phenyl, Y represents oxygen, and Z represents NH.  
     
     
         104 . The device of  claim 102  wherein the second reactive intermediate has a formula vii: 
         [(R 1 )(R 2 )(R 3 )Si—X-Z-CYNH] 2 Si [NHCYL-M] 2    vii 
       wherein R 1 , R 2  and R 3  independently represents C 1 -C 6  alkoxy, C 1 -C 6  alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy; L represents a linking group; X represents linear or branched C 1 -C 20  alkyl or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; and Z represents oxygen or NH; and M represents a molecule, with the proviso that at least one of R 1 , R 2 , or R 3  represent C 1 -C 6  alkoxy.  
     
     
         105 . The device of  claim 104  wherein L represents a nucleophilic group from the molecule.  
     
     
         106 . The device of  claim 105  wherein the nucleophilic group comprises —NH, —S—, —O—, or —OOC—.  
     
     
         107 . The device of  claim 57  wherein the molecule comprises a probe.  
     
     
         108 . The device of  claim 107 , wherein the probe comprises a protein, a peptide, a nucleic acid, a peptide nucleic acid, an amino acid, a linked nucleic acid, a nucleoside triphosphate, a carbohydrate, a lipid, a lipid bound protein, an aptamer, a virus, a cell fragment, or a whole cell.  
     
     
         109 . The device of  claim 108 , wherein the lipid bound protein comprises a G-protein coupled receptor.  
     
     
         110 . The device of  claim 107 , wherein the probe comprises an antibody, an antigen, a receptor, or a ligand.  
     
     
         111 . A device for immobilizing a molecule onto a glass surface, wherein the molecule is immobilized by the method comprising the steps of: 
 (a) contacting Si(NCY) 4  wherein Y represents oxygen or sulfur with an agent so as to form a first reactive intermediate, said agent having a formula ii:     (R 1 )(R 2 )(R 3 )Si—X-Z  ii   wherein R 1 , R 2  and R 3  independently represents C 1 -C 6  alkoxy, C 1 -C 6  alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy; X represents linear or branched C 1 -C 20  alkyl or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, and sulfur; and Z represents a hydroxy or amino group, with the provision that at least of one R 1 , R 2 , or R 3  represents C 1 -C 6  alkoxy;    (b) contacting the first reactive intermediate with a molecule so as to form a second reactive intermediate;    (c) contacting the second reactive intermediate with said surface so as to immobilized the molecule onto said surface.    
     
     
         112 . The device of  claim 111  wherein the surface is a glass surface.  
     
     
         113 . The device of  claim 111  wherein the surface has at least one group that reacts with the reactive intermediate.  
     
     
         114 . The device of  claim 113  wherein the group comprises a hydroxyl or amino group.  
     
     
         115 . The device of  claim 111  wherein the agent is 4-aminophenyl trimethoxysilane, 4-aminophenyl trimethoxysilane, 3-aminophenyltriethoxysilane, 3-aminophenyltriethoxysilane, or mixture thereof.  
     
     
         116 . The device of  claim 111  wherein the agent and tetraisocyanatosilane is present at a 1:1 molar ratio.  
     
     
         117 . The device of  claim 111  wherein the molecule and the first reactive intermediate is present in a 3:1 molar ratio.  
     
     
         118 . The device of  claim 111  wherein the first reactive intermediate has a formula iv: 
         (R 1 )(R 2 )(R 3 )Si—X-Z-CYNH—Si (NCY) 3   iv 
       wherein R 1 , R 2  and R 3  independently represents C 1 -C 6  alkoxy, C 1 -C 6  alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy; X represents linear or branched C 1 -C 20  alkyl or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; and Z represents oxygen or NH, with the proviso that at least one of R 1 , R 2 , or R 3  represents C 1 -C 6  alkoxy.  
     
     
         119 . The device of  claim 118  wherein R 1 , R 2  and R 3  represents methoxy, X represents phenyl, Y represents oxygen, and Z represents NH.  
     
     
         120 . The device of  claim 118  wherein the second reactive intermediate has a formula v: 
         (R 1 )(R 2 )(R 3 )Si—X-Z-CYNH—Si (NHCYL-M) 3   v 
       wherein R 1 , R 2  and R 3  independently represents C 1 -C 6  alkoxy, C 1 -C 6  alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy; L represents a linking group; X represents linear or branched C 1 -C 20  alkyl or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; and Z represents oxygen or NH; and M represents a molecule, with the proviso that at least one of R 1 , R 2 , or R 3  represents C 1 -C 6  alkoxy.  
     
     
         121 . The device of  claim 120  wherein L represents a nucleophilic group from the molecule.  
     
     
         122 . The device of  claim 121  wherein the nucleophilic group comprises —NH, —S—, —O—, or —OOC—.  
     
     
         123 . The device of  claim 111  wherein the first reactive intermediate has a formula vi: 
         [(R 1 )(R 2 )(R 3 )Si—X-Z-CYNH] 2 —Si (NCY) 2   vi 
       wherein R 1 , R 2  and R 3  independently represents C 1 -C 6  alkoxy, C 1 -C 6  alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy; X represents linear or branched C 1 -C 20  alkyl or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; and Z represents oxygen or NH, with the proviso that at least one of R 1 , R 2 , or R 3  represents C 1 -C 6  alkoxy.  
     
     
         124 . The device of  claim 123  wherein R 1 , R 2  and R 3  represents methoxy, X represents phenyl, Y represents oxygen, and Z represents NH.  
     
     
         125 . The device of  claim 123  wherein the second reactive intermediate has a formula vii: 
         [(R 1 )(R 2 )(R 3 )Si—X-Z-CYNH] 2 Si (NHCYL-M) 2   vii 
       wherein R 1 , R 2  and R 3  independently represents C 1 -C 6  alkoxy, C 1 -C 6  alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy; L represents a linking group; X represents linear or branched C 1 -C 20  alkyl or aryl substituted With one or more groups selected from the group consisting of C 1 -C 6  alkyl and C 1 -C 6  alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; and Z represents oxygen or NH; and M represents a molecule, with the proviso that at least one of R 1 , R 2 , or R 3  represents C 1 -C 6  alkoxy.  
     
     
         126 . The device of  claim 125  wherein L represents a nucleophilic group from the molecule.  
     
     
         127 . The device of  claim 126  wherein the nucleophilic group comprises —NH, —S—, —O—, or —OOC—.  
     
     
         128 . The device of  claim 111  wherein the molecule comprises a probe.  
     
     
         129 . The device of  claim 128 , wherein the probe comprises a protein, a peptide, a nucleic acid, a peptide nucleic acid, an amino acid, a linked nucleic acid, a nucleoside triphosphate, a carbohydrate, a lipid, a lipid bound protein, an aptamer, a virus, a cell fragment, or a whole cell.  
     
     
         130 . The device of  claim 129 , wherein the lipid bound protein comprises a G-protein coupled receptor.  
     
     
         131 . The device of  claim 111 , wherein the probe comprises an antibody, an antigen, a receptor, or a ligand.