Preparation of nanometric arrays of biomolecules on oligo- or poly(ethylene glycol) films on silicon surfaces
The present invention is generally directed to nanometric biomolecular arrays and to a novel approaches for the preparation of such nanoarrays, based on binding of biomolecules, such as avidin, to templates generated by lithographically-anodizing biocompatible ultrathin films on silicon substrates using AFM anodization lithography. The present invention is also directed to methods of using such arrays.
1. A method comprising the steps of:
a) contacting OEG-terminated alkenes with a hydrogen-terminated Si surface to form a contacted surface; wherein the OEG-terminated alkenes comprise α-heptyl-(ethylene glycol) methyl ω-undecenyl ether, wherein the Si surface is selected from the group consisting of Si(100), Si(111), and combinations thereof;
b) photolyzing the contacted surface to effect Si—C bonding between the OEG-terminated alkenes and the Si surface and form a OEG-coated Si surface comprising a monolayer of OEG bound to the Si surface through Si—C bonds, wherein the monolayer of OEG resists non-specific adsorption of biomolecules with greater than 99% reduction in adsorption with respect to the Si surface;
c) lithographically anodizing the head groups of the OEG monolayer of the OEG-coated Si surface via AFM anodization lithography to yield a nanolithographically-patterned OEG-coated Si surface, wherein the nanolithographically-patterned regions are further etched to form nanoholes, wherein the nanoholes comprise regions with enhanced associability toward biomolecules, wherein the nanolithographically-patterned regions resist denaturing of biomolecules; and
d) depositing at least one type of biomolecule in the nanolithographically-patterned regions of enhanced associability to form a nanometric biomolecular array.
2. The method of claim 1 , wherein the Si surface is atomically flat.
3. The method of claim 1 , further comprising a step of washing the OEG-coated Si surface prior to lithographically anodizing regions on top of it.
4. The method of claim 1 , wherein the nanolithographically-patterned regions of the OEG-coated Si surface comprise nanowells.
5. The method of claim 1 , wherein the nanolithographically-patterned regions comprise functional moieties selected from the group consisting of carboxylic acid, aldehyde, hydroxyl, and combinations thereof.
6. The method of claim 5 , wherein the functional moieties provide, at least in part, the enhanced associability toward biomolecules.
7. The method of claim 1 , wherein the nanolithographically patterned regions of the OEG-coated Si surface comprise hydroxyl moieties that can be converted into carboxylic acid moieties.
8. The method of claim 1 , wherein the at least one type of biomolecule is selected from the group consisting of proteins, oligonucleotides, and combinations thereof.
9. The method of claim 1 , wherein the at least one type of biomolecule comprises avidin.
10. The method of claim 1 , wherein at least some of the at least one type of biomolecule binds with the regions with enhanced associability via amide bonds.
11. The method of claim 1 , further comprising the steps of:
a) depositing biomolecular analyte; and
b) analyzing the array to determine the regions in which the biomolecular analyte exhibits a binding affinity.
12. The method of claim 11 , further comprising a step of regenerating the nanolithographically-patterned OEG-coated Si surface comprising regions with enhanced associability toward biomolecules by treatment of said surface with proteinase K, wherein the proteinase K serves to catalyze hydrolytic fragmentation of proteins bound to said surface.