Molecular microarrays and helical peptides

Abstract

Methods for fabricating dense arrays of polymeric molecules in a highly multiplexed manner are provided using semiconductor-processing-derived lithographic methods. Advantageously, the methods are adaptable to the synthesis of a variety of polymeric compounds. For example, arrays of branched peptides and polymers joined by peptide bonds may be fabricated in a highly multiplexed manner. Additionally, peptides that adopt helical structures are synthesized on a substrate surface and arrays are created having one or more features containing peptides capable of forming helixes.

1 . A method for making a peptide on a substrate comprising,
 attaching to a region of a substrate surface a trifunctional linker molecule having three functional groups each of which is capable of forming a bond, where in the trifunctional linker is attached to the region of the substrate surface through one of the three functional groups, wherein the two remaining functional groups of the trifunctional linker molecule are each capable of forming a peptide bond and wherein the trifunctional linker molecule comprises two different protecting groups that prevent the formation of a peptide bond by the two remaining functional groups,   removing selectively one of the protecting groups of the trifunctional linker molecule,   coupling a second molecule having two functional groups that are each capable of forming a peptide bond to the deprotected trifunctional linker molecule, wherein the second molecule comprises a protecting group capable of preventing the formation of a peptide bond, to a functional group of the trifunctional linker molecule that has been deprotected,   removing selectively a second one of the protecting groups of the trifunctional linker molecule, and   coupling a third molecule having two functional groups that are each capable of forming a peptide bond to the deprotected trifunctional linker molecule, wherein the second molecule comprises a protecting group capable of preventing the formation of a peptide bond, to a functional group of the trifunctional linker molecule that has been deprotected.   
   
   
       2 . The method according to  claim 1  also including capping any unreacted deprotected functional groups on the trifunctional linker molecule after coupling the second molecule capable of forming two peptide bonds. 
   
   
       3 . The method according to  claim 1  wherein the substrate is selected from the group consisting of glass, silicon, and silicon having a SiO 2  layer. 
   
   
       4 . The method according to  claim 1  wherein a protecting group is t-butoxycarbonyl, benzyloxycarbonyl, or 9-fluorenylmethoxycarbonyl. 
   
   
       5 . The method according to  claim 1  wherein coupling a molecule capable of forming two peptide bonds is repeated a plurality of times forming a peptide attached to the substrate surface having a size from 4 peptide bonds to 25 peptide bonds. 
   
   
       6 . The method according to  claim 1  wherein coupling a molecule capable of forming two peptide bonds is repeated a plurality of times forming a peptide attached to the substrate surface that is capable of forming a helical structure. 
   
   
       7 . The method according to  claim 6  wherein the peptide attached to the substrate surface that is capable of forming a helical structure comprises one or more heptad repeats. 
   
   
       8 . The method according to  claim 1  wherein the region of the substrate is one of a plurality of regions, wherein the regions comprise an array of regions, and wherein a plurality of the plurality of regions comprise peptides. 
   
   
       9 . The method according to  claim 8  wherein the area occupied by a region of the array is less than 1 mm 2 . 
   
   
       10 . The method according to  claim 8  wherein the area occupied by a region of the array is less than 1 μm 2 . 
   
   
       11 . The method according to  claim 8  wherein the array comprises 1,000 to 10,000 regions.