Method of patterning molecules on a substrate using a micro-contact printing process

Abstract

The present invention relates to a method of patterning molecules on a substrate using a micro-contact printing process, to a substrate produced by said method and to uses of said substrate. It also relates to a device for performing the method according to the present invention.

1 - 30 . (canceled) 
     
     
         31 . A substrate comprising a pattern of molecules thereon, which molecules have retained their function and/or activity and/or native conformation, the substrate produced by a method of patterning the molecules on the substrate using a micro-contact printing process, whereby the molecules to be patterned are kept in solvent or are covered by solvent during the entire micro-contact printing process, said method comprising:
 providing molecules to be patterned in a solvent and providing a patterned surface, said molecules being first immobilized on an ink-pad within said solvent;   transferring said molecules to be patterned onto said patterned surface and immobilizing them thereon while keeping said molecules in said solvent or covered by said solvent, said ink-pad having said molecules immobilized thereon is brought into conformal contact with said patterned surface in a first solvent environment containing said molecules to be patterned and said solvent, thereby transferring said molecules onto said patterned surface and immobilizing them thereon; and   providing said substrate in a second solvent environment, transferring said patterned surface having said molecules immobilized thereon to said second solvent environment, and bringing said patterned surface having said molecules immobilized thereon into conformal contact with said substrate, thereby creating a pattern of said molecules on said substrate, while keeping said molecules in said solvent or covered by said solvent, wherein said molecules to be patterned are protein molecules,   wherein said micro-contact printing process occurs in the absence of a drying step.   
     
     
         32 . The substrate according to  claim 31 , wherein the bringing into conformal contact of said patterned surface with said substrate of the providing a substrate occurs in a period not longer than 180 min after immobilizing said molecules to be patterned on said patterned surface. 
     
     
         33 . The substrate according to  claim 31 , wherein, after the providing a substrate, said patterned surface is lifted from said substrate, thereby leaving behind a substrate having a pattern of said molecules thereon. 
     
     
         34 . The substrate according to  claim 33 , wherein said substrate having a pattern of molecules thereon is kept in or covered by solvent containing a buffer. 
     
     
         35 . The substrate according to  claim 31 , wherein said molecules to be patterned retain their function and/or activity and/or native conformation throughout the entire process, due to their being kept in solvent or covered by solvent during the entire micro-contact printing process. 
     
     
         36 . The substrate according to  claim 31 , further comprising a spacer layer and/or a binding layer which facilitates binding of said substrate to said molecules to be patterned through covalent binding, electrostatic forces, van der Waals forces, H-bonding, London forces or any combination of the foregoing. 
     
     
         37 . The substrate according to  claim 31 , selected from the group consisting of: metals and semi metals, single or polycrystalline materials; single or polycrystalline metals and semi metals, gold, platinum, or silicon, or; composite materials of single or polycrystalline composites, siliconoxide, or GaAs, or amorphous composite materials or glass; plastics, or elastomers or polydimethylsiloxane, or plastomers, or polyolefines, or ionomers, or resist materials, or UV-NIL resists. 
     
     
         38 . The substrate according to  claim 31 , wherein said patterned surface is made from a material selected from the group consisting of: single-crystalline materials and polycrystalline materials, silicon, silicon oxide, layered composite systems, silicon oxide on silicon, metal layers on silicon or metal layers on silicon oxide; amorphous materials, glass; plastics, elastomers, polydimethylsiloxane, plastomers, polyolefines (POP, polyolefinic plastomers), ionomers, resist materials, or UV-NIL-resists. 
     
     
         39 . The substrate according to  claim 31 , wherein at least a surface of said ink-pad is made from a material selected from the group consisting of: single-crystalline materials and polycrystalline materials, silicon, silicon oxide, layered composite systems, silicon oxide on silicon, metal layers on silicon/siliconoxide; amorphous materials, glass; plastics, elastomers, polydimethylsiloxane, plastomers, polyolefines (POP, polyolefinic plastomers), ionomers, resist materials, or UV-NIL-resists. 
     
     
         40 . The substrate according to  claim 31 , wherein said molecules to be patterned are selected from the group consisting of: redox proteins, nucleic-acid binding proteins, metallo-proteins, cytochrome c, azurin, cytoskeleton-proteins. 
     
     
         41 . The substrate according to  claim 31 , wherein said molecules to be patterned have one or plural lysine residues, and wherein said substrate is Au, or Au having a spacer layer on its surface so as to avoid denaturation of said protein, said spacer layer having a thickness in the range of from 0.5 nm to 200 nm. 
     
     
         42 . The substrate according to  claim 31 , wherein the pattern comprises features having a length in the range of from approximately 10 nm to 500 μm. 
     
     
         43 . The substrate according to  claim 31 , wherein said providing a patterned surface includes providing a patterned surface in the form of a stamp. 
     
     
         44 . The substrate according to  claim 31 , wherein said providing molecules to be patterned in a solvent includes providing molecules first immobilized on an ink-pad in the form of a non-patterned surface within said solvent. 
     
     
         45 . The substrate according to  claim 31 , wherein said first solvent environment also includes a buffer. 
     
     
         46 . The substrate according to  claim 31 , wherein said second solvent environment also includes a buffer. 
     
     
         47 . The substrate according to  claim 31 , wherein the bringing into conformal contact of said patterned surface with said substrate of the providing a substrate occurs in a period not longer than 120 min after immobilizing said molecules to be patterned on said patterned surface. 
     
     
         48 . The substrate according to  claim 31 , wherein the bringing into conformal contact of said patterned surface with said substrate of the providing a substrate occurs in a period not longer than 10 min after immobilizing said molecules to be patterned on said patterned surface. 
     
     
         49 . The substrate according to  claim 31 , wherein the bringing into conformal contact of said patterned surface with said substrate of the providing a substrate occurs in a period not longer than 1 min after immobilizing said molecules to be patterned on said patterned surface. 
     
     
         50 . The substrate according to  claim 31 , wherein the pattern comprises features having a length in the range of from approximately 10 nm to ≦200 nm. 
     
     
         51 . The substrate according to  claim 31 , wherein the pattern comprises features having a length in the range of from approximately 10 nm to ≦150 nm. 
     
     
         52 . The substrate according to  claim 31 , wherein the substrate is modified with a molecular layer. 
     
     
         53 . The substrate of  claim 52 , wherein the molecular layer is a self assembling monolayer (SAM). 
     
     
         54 . The substrate of  claim 53 , wherein the molecules of the SAM have two termini, a first terminus for binding to the substrate, and a second terminus for binding the protein molecules to be patterned. 
     
     
         55 . The substrate of  claim 54 , wherein the substrate comprises gold and the first terminus comprises a thiol for binding to the gold. 
     
     
         56 . The substrate of  claim 54 , wherein the substrate comprises silicon oxide and the first terminus comprises a silane for binding to the silicon oxide. 
     
     
         57 . The substrate according to  claim 54 , wherein the SAM is selected from the group consisting of: a SAM with mercapto- or amino-groups for binding metals, a SAM with carboxy-groups for electrostatic binding, a SAM with mercapto-groups for binding metals, with plain alkyl chains having methylene groups for van der Waals interaction, with —COOH, —OH or vinyl-groups for covalent coupling, or SAMs with antibodies for binding corresponding antigens, or SAMs with antigens for binding corresponding antibodies, or SAMs with receptors for specific binding of molecules. 
     
     
         58 . The substrate according to  claim 52 , wherein the molecular layer includes antibodies for binding antigens or includes antigens for binding corresponding antibodies. 
     
     
         59 . The substrate according to  claim 52 , wherein the molecular layer includes receptors for specific binding of molecules. 
     
     
         60 . The substrate according to  claim 31 , wherein the substrate is gold modified with a mercapto undecanoic acid layer (MUA).