Arrays of protein-capture agents and methods of use thereof

Abstract

Arrays of protein-capture agents useful for the simultaneous detection of a plurality of proteins which are the expression products, or fragments thereof, of a cell or population of cells in an organism are provided. A variety of antibody arrays, in particular, are described. Methods of both making and using the arrays of protein-capture agents are also disclosed. The invention arrays are particularly useful for various proteomics applications including assessing patterns of protein expression and modification in cells.

1 .- 45 . (canceled)  
     
     
         46 . A biochip comprising: 
 a substrate; and    an array of regions on the substrate, wherein the regions in the array of regions comprise protein capture agents, wherein the protein capture agents comprise aptamers.    
     
     
         47 . The biochip of  claim 46  wherein the protein capture agents consist of aptamers.  
     
     
         48 . The biochip of  claim 46  further comprising proteins bound to the aptamers.  
     
     
         49 . The biochip of  claim 46  wherein the regions in the array of regions have a density of at least about 100 regions per cm 2 .  
     
     
         50 . The biochip of  claim 46  wherein the aptamers are capable of binding to proteins.  
     
     
         51 . The biochip of  claim 46  wherein the aptamers are DNA aptamers.  
     
     
         52 . A method for making an biochip, the method comprising: 
 obtaining a substrate; and    forming an array of regions on the substrate wherein the regions in the array of regions comprise protein capture agents wherein the protein capture agents comprise aptamers.    
     
     
         53 . The method of  claim 52  wherein the protein capture agents consist of aptamers.  
     
     
         54 . The method of  claim 52  wherein the regions in the array of regions have a density of at least about 100 regions per cm 2 .  
     
     
         55 . The method of  claim 52  wherein the aptamers are capable of binding to proteins.  
     
     
         56 . The method of  claim 52  wherein the array of regions contains more than 1000 regions.  
     
     
         57 . The method of  claim 52  further comprising forming a polymer thin film on the substrate.  
     
     
         58 . A method of assaying, the method comprising: 
 obtaining a substrate and an array of regions on the substrate, the regions in the array of regions comprising protein capture agents comprising aptamers;    introducing a fluid to the substrate, the fluid comprising proteins;    detecting the binding, if any, of the proteins to the aptamers.    
     
     
         59 . The method of  claim 58  wherein the proteins are human proteins.  
     
     
         60 . The method of  claim 58  wherein the regions in the array of regions have a density of at least about 100 regions per cm 2 .  
     
     
         61 . The method of  claim 58  wherein detecting comprises using an optical detection process to detect any binding to the aptamers.  
     
     
         62 . The method of  claim 58  wherein the aptamers are DNA aptamers.  
     
     
         63 . The method of  claim 46 , wherein the protein capture agents are photocrosslinked to analytes.  
     
     
         64 . A biochip comprising: 
 a substrate; and    an array of regions on the substrate, at least one region comprising a first polynucleotide hybridized to a second polynucleotide, and an analyte bound to the second polynucleotide.    
     
     
         65 . The biochip of  claim 64  wherein the analyte is a protein.  
     
     
         66 . The biochip of  claim 64  wherein the regions comprise respectively different proteins.  
     
     
         67 . The biochip of  claim 64  wherein the second polynucleotide facilitates attachment of the analyte to the first polynucleotide.  
     
     
         68 . The biochip of  claim 64  wherein the second polynucleotide facilitates attachment of the analyte to the first polynucleotide via a capture agent.