Method for attaching biological molecules to a glass surface
The invention describes a method for attaching biological molecules, such as proteins, nucleic acids, and carbohydrates to a glass surface. In particular, a covalent attachment method is described wherein a glass surface is first activated with a water-stable form of an isocyanate, termed a protected isocyanate. The protecting group of the isocyanate moiety is displaced by amines, hydroxyl, or carboxyl groups of biological molecules, leading to covalent attachment to the glass surface.
1. A method for attaching a biological molecule to a glass surface comprising the steps of:
a) providing a silane-treated glass surface having an end-capped amino group;
b) reacting said end-capped amino group with a phosgene equivalent to form a protected isocyanate group; and
c) reacting said protected isocyanate group with a biological molecule,
thereby forming a covalent coupling and attaching said biological molecule to the glass surface.
2. The method of claim 1 further comprising reacting a glass surface with an aminosilane thereby forming the silane-treated glass surface having an end-capped amino group.
3. The method of claim 1 wherein the phosgene equivalent is a carbonyl diimidazole or a ketoxime carbonate.
4. The method of claim 3 wherein the carbonyl diimidazole is 1,1-carbonyldiimidazole.
5. The method of claim 3 wherein the ketoxime carbonate is methyl ethyl ketoxime carbonate.
6. The method of claim 1 wherein the biological molecule is selected from the group consisting of proteins, peptides, nucleic acids and carbohydrates.
7. The method of claim 1 wherein the glass surface is a microscope slide surface.
8. A method for attaching a biological molecule to a glass surface comprising the steps of:
a) providing a silane-treated glass surface having an end-capped amino group;
b) reacting said amino group with a phosgene equivalent to form an end-capped group, said end-capped group including a functional group represented by the following structural formula:
>N—C(O)—N<
or by the following structural formula:
>N—C(O)—O—N<;
and
c) reacting said end-capped group with a biological molecule, thereby forming a covalent coupling and attaching said biological molecule to the glass surface.
9. The method of claim 8 wherein the phosgene equivalent is a carbonyl diimidazole or a ketoxime carbonate.
10. The method of claim 9 wherein the carbonyl diimidazole is 1,1-carbonyldiimidazole.
11. The method of claim 9 wherein the ketoxime carbonate is methyl ethyl ketoxime carbonate.
12. The method of claim 8 wherein said end-capped group is represented by the following structural formula:
13. The method of claim 8 wherein said end-capped group is represented by the following structural formula:
14. A method for attaching a biological molecule to a silane-treated glass surface comprising the steps of:
a) reacting an end-capped amino group on said silane-treated glass surface with a phosgene equivalent to form a protected isocyanate group; and
b) reacting said protected isocyanate group with a biological molecule,
thereby forming a covalent coupling and attaching said biological molecule to said silane-treated glass surface.
15. A method for attaching a biological molecule to a silane-treated glass surface comprising the steps of:
a) reacting an amino group on said silane-treated glass surface with a phosgene equivalent to form an end-capped group, said end-capped group including a functional group represented by the following structural formula:
>N—C(O)—N<
or by the following structural formula:
>N—C(O)—O—N<;
and
b) reacting said end-capped group with a biological molecule, thereby forming a covalent coupling and attaching said biological molecule to said silane-treated glass surface.
16. The method of claim 8 wherein the glass surface is a microscope slide surface.