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 . A system for producing a substrate including a pattern of molecules thereon, which molecules have retained their function and/or activity and/or native conformation, the system comprising:
 a first container including an ink-pad, molecules to be patterned, and a first solvent covering the ink-pad and the molecules, the molecules being immobilized on the ink-pad within the first solvent;   a patterned surface onto which the molecules to be patterned are transferred and immobilized therein while keeping the molecules in the first solvent and covered by the first solvent, by bringing the ink-pad having the molecules immobilized thereon into conformal contact with the patterned surface in the first container to transfer the molecules onto the patterned surface and immobilizing the molecules on the patterned surface; and   a second container including a second solvent and a substrate covered by the second solvent, the patterned surface having the molecules immobilized thereon being brought into conformal contact with the substrate within the second solvent, thereby creating a pattern of the molecules on the substrate, while keeping the molecules in the second solvent and covered by the second solvent;   wherein the molecules to be patterned are protein molecules.   
     
     
         2 . The system according to  claim 1 , wherein the patterned surface is brought into conformal contact with the substrate for a period not longer than 180 min after immobilizing the molecules to be patterned on the patterned surface. 
     
     
         3 . The system according to  claim 1 , wherein, after the patterned surface is brought into conformal contact with the substrate, the patterned surface is lifted from the substrate, thereby leaving behind a substrate having a pattern of the molecules thereon. 
     
     
         4 . The system according to  claim 3 , wherein the substrate having the pattern of molecules thereon is kept in or covered by the second solvent which contains a buffer. 
     
     
         5 . The system according to  claim 1 , wherein the molecules to be patterned retain their function and/or activity and/or native conformation due to their being kept in solvent or covered by solvent. 
     
     
         6 . The system according to  claim 1 , further comprising in the second container a spacer layer and/or a binding layer that facilitates binding of the substrate to the molecules to be patterned through covalent binding, electrostatic forces, van der Waals forces, H-bonding, London forces, or any combination of the foregoing. 
     
     
         7 . The system according to  claim 1 , wherein the substrate is 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. 
     
     
         8 . The system according to  claim 1 , wherein the 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.   
     
     
         9 . The system according to  claim 1 , wherein at least a surface of the 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. 
     
     
         10 . The system according to  claim 1 , wherein the molecules to be patterned are selected from the group consisting of: redox proteins, nucleic-acid binding proteins, metallo-proteins, cytochrome c, azurin, cytoskeleton-proteins. 
     
     
         11 . The system according to  claim 1 , wherein the molecules to be patterned have one or plural lysine residues, and wherein the substrate is Au, or Au having a spacer layer on its surface so as to avoid denaturation of the protein, the spacer layer having a thickness in the range of from 0.5 nm to 200 nm. 
     
     
         12 . The substrate according to  claim 1 , wherein the pattern comprises features having a length in a range of from approximately 10 nm to 500 μm. 
     
     
         13 . The substrate according to  claim 1 , wherein the patterned surface is in a form of a stamp. 
     
     
         14 . The substrate according to  claim 1 , wherein the molecules are first immobilized on the ink-pad in a form of a non-patterned surface within the solvent. 
     
     
         15 . The system according to  claim 1 , wherein the first solvent environment includes a buffer. 
     
     
         16 . The system according to  claim 1 , wherein the second solvent environment includes a buffer. 
     
     
         17 . The system according to  claim 1 , wherein the patterned surface is brought into conformal contact with the substrate for a period not longer than 120 min after immobilizing the molecules to be patterned on the patterned surface. 
     
     
         18 . The system according to  claim 1 , wherein the patterned surface is brought into conformal contact with the substrate for a period not longer than 10 min after immobilizing the molecules to be patterned on the patterned surface. 
     
     
         19 . The system according to  claim 1 , wherein the patterned surface is brought into conformal contact with the substrate for a period not longer than 1 min after immobilizing the molecules to be patterned on the patterned surface. 
     
     
         20 . The system according to  claim 19 , wherein the substrate is modified with a molecular layer. 
     
     
         21 . The system of  claim 19 , wherein the molecular layer is a self assembling monolayer (SAM). 
     
     
         22 . The system of  claim 21 , 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. 
     
     
         23 . The system of  claim 22 , wherein the substrate comprises gold and the first terminus comprises a thiol for binding to the gold. 
     
     
         24 . The system of  claim 22 , wherein the substrate comprises silicon oxide and the first terminus comprises a silane for binding to the silicon oxide. 
     
     
         25 . The system according to  claim 22 , 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. 
     
     
         26 . The system according to  claim 20 , wherein the molecular layer includes antibodies for binding antigens or includes antigens for binding corresponding antibodies. 
     
     
         27 . The system according to  claim 20 , wherein the molecular layer includes receptors for specific binding of molecules. 
     
     
         28 . The system according to  claim 20 , wherein the substrate is gold modified with a mercapto undecanoic acid layer (MUA).