Polypeptide Immobilization

Abstract

The present invention provides a method, comprising (a) providing a reactant ligand attached to a substrate; (b) contacting the substrate with a fusion polypeptide, said fusion polypeptide comprising a capture polypeptide fused to a display polypeptide under conditions such that said reactant ligand covalently binds to said capture polypeptide; and (c) analyzing said display polypeptide.

1 - 15 . (canceled) 
     
     
         16 . A system, comprising:
 (a) a plurality of hydrolase reactant ligands for a hydrolase attached to a substrate;   (b) a plurality of fusion polypeptides, wherein each of said fusion polypeptides comprises a hydrolase capture polypeptide fused to a display polypeptide, wherein the hydrolase capture polypeptide is capable of covalently binding to said hydrolase reactant ligands.   
     
     
         17 . The system of  claim 16 , wherein said substrate comprises at least one member selected from the group consisting of metal, metal oxide, glass, ceramic, quartz, silicon, polymer, sepharose, agarose, a colloid, a lipid bilayer, and a lipid monolayer. 
     
     
         18 . The system of  claim 16 , wherein said display polypeptide comprises a polypeptide of interest to be immobilized to said substrate. 
     
     
         19 . The system of  claim 16 , wherein said hydrolase capture polypeptide comprises cutinase. 
     
     
         20 . The system of  claim 16 , wherein the plurality of hydrolase reactant ligands are organized into an array. 
     
     
         21 . A protein chip comprising the system of  claim 16 , wherein the hydrolase capture polypeptides of the plurality of fusion polypeptides are covalently bound to said hydrolase reactant ligands. 
     
     
         22 . The protein chip of  claim 21 , wherein said substrate comprises at least one member selected from the group consisting of metal, metal oxide, glass, ceramic, quartz, silicon, polymer, sepharose, agarose, a colloid, a lipid bilayer, and a lipid monolayer. 
     
     
         23 . The protein chip of  claim 21 , wherein said display polypeptide comprises a polypeptide of interest to be immobilized to said substrate. 
     
     
         24 . The protein chip of  claim 21 , wherein said hydrolase capture polypeptide comprises cutinase. 
     
     
         25 . The protein chip of  claim 21 , wherein the plurality of hydrolase reactant ligands and the fusion polypeptides covalently attached thereto are organized into an array. 
     
     
         26 . A vector for use in producing the system of  claim 16 , said vector comprising:
 (a) a promoter;   (b) a hydrolase capture polypeptide coding region under transcriptional control of said promoter; and   (c) a cloning site, in-frame with said hydrolase capture polypeptide coding region, for inserting a nucleic acid encoding a display polypeptide.   
     
     
         27 . The vector of  claim 26 , wherein transcription and translation of said vector produces a fusion polypeptide comprising said hydrolase capture polypeptide and said display polypeptide. 
     
     
         28 . The vector of  claim 26 , further comprising:
 (d) a selectable marker to facilitate identifying cells that have incorporated the vector.   
     
     
         29 . The vector of  claim 28 , wherein said selectable marker comprising an antibiotic resistance gene. 
     
     
         30 . A surface for use in producing the system of  claim 16 , said surface comprising plurality of hydrolase reactant ligands attached to a substrate, wherein upon interacting with said hydrolase capture polypeptides, covalent bonds are formed between said hydrolase capture polypeptides and said hydrolase reactant ligands. 
     
     
         31 . The surface of  claim 30 , wherein said substrate comprises at least one member selected from the group consisting of metal, metal oxide, glass, ceramic, quartz, silicon, polymer, sepharose, agarose, a colloid, a lipid bilayer, and a lipid monolayer.