Detection and assay devices and methods of making and using the same

Abstract

An article such as a biomolecular detector or biosensor having a nonfouling surface thereon includes: (a) a substrate having a surface portion; (b) a linking layer on the surface portion; and (c) a polymer layer formed on the linking layer; and (d) a first member of a specific binding pair (e.g., a protein, peptide, antibody, nucleic acid, etc.) bound to the polymer layer. Methods of making and using the articles are also described.

1 - 20 . (canceled) 
     
     
         21 . A method of making a biomolecular detector, said method comprising:
 (a) providing a substrate having a surface portion, a linking layer on said surface portion, and a polymer layer comprised of monomeric units layer formed on said linking layer, with each of said monomeric units comprising a monomer core group having at least one protein-resistant head group coupled thereto, to thereby form a brush molecule on said surface portion; said brush molecule comprising a stem formed from the polymerization of said monomer core groups, and a plurality of branches formed from said hydrophilic head group projecting from said stem; and then   (b) non-covalently bonding a member of a specific binding pair to said polymer layer.   
     
     
         22 . The method of  claim 21 , wherein said brush molecule is from 5 to 200 nanometers in length. 
     
     
         23 . The method of  claim 21 , said brush molecule formed on said surface portion at a density from 10 to 500 milligrams per meter 2 . 
     
     
         24 . The method of  claim 21 , wherein said first member is bonded to said polymer layer at a density of from 1 milligram per meter 2  to 50 grams per meter 2 . 
     
     
         25 . The method of  claim 21 , wherein said providing step comprises providing said substrate in macroscopically dry form. 
     
     
         26 . The method of  claim 21 , wherein said providing step comprises providing said substrate in combination with water, said water consisting essentially of water of hydration bonded to said polymer layer, and with said substrate in macroscopically dry form. 
     
     
         27 . The method of  claim 21 , wherein said non-covalent bonding step is carried out by depositing an aqueous solution containing said first member of a specific binding pair on said polymer layer. 
     
     
         28 . The method of  claim 21 , wherein said first member is a protein, peptide, or nucleic acid. 
     
     
         29 . The method of  claim 21 , wherein said first member is an antibody. 
     
     
         30 . The method of  claim 21 , further comprising the step of: (c) drying said polymer layer following said non-covalently bonding step. 
     
     
         31 . The method of  claim 21 , further comprising the step of: (d) sealing said substrate in macroscopically dry form in a container. 
     
     
         32 . The method of  claim 21 , wherein: said first member of said specific binding pair is non-covalently bound to said polymer at a discrete probe location, said method further comprising: (c) non-covalently bonding a plurality of additional first members of a specific binding pairs to said polymer layer at a plurality of additional discrete probe locations to thereby form an array thereon. 
     
     
         33 . The method of  claim 32 , wherein said discrete probe locations are formed on said polymer layer at a density of 5 to 10,000 discrete probe locations per cm 2 . 
     
     
         34 . The method of  claim 32 , wherein said discrete probe locations are formed on said polymer layer at a density of 10,000 to 1 million discrete probe locations per cm 2 . 
     
     
         35 . The method of  claim 32 , wherein said discrete probe locations are formed on said polymer layer at a density of 1 million to 1 billion discrete probe locations per cm 2 .