Non-fouling polymeric surface modification and signal amplification method for biomolecular detection

Patent metadata

Field	Value
Publication number	US-9890420-B2
Application number	US-201615218625-A
Country	US
Kind code	B2
Filing date	Jul 25, 2016
Priority date	Sep 15, 2005
Publication date	Feb 13, 2018
Grant date	Feb 13, 2018

Abstract

An article such as a biosensor having a nonfouling surface thereon is described. The article comprises: (a) a substrate having a surface portion; (b) a linking layer on the surface portion; (c) a polymer layer comprising brush molecules formed on the linking layer; and (d) optionally but preferably, a first member of a specific binding pair (e.g., a protein, peptide, antibody, nucleic acid, etc.) coupled to the brush molecules. The polymer layer is preferably formed by the process of surface-initiated polymerization (SIP) of monomeric units thereon. Preferably, each of the monomeric units comprises a monomer (for example, a vinyl monomer) core group having at least one protein-resistant head group coupled thereto, to thereby form the brush molecule on the surface portion. Methods of using the articles are also described.

First claim

That which is claimed is:

1. A biomolecular detector, comprising:
(a) a substrate having a surface portion;
(b) a linking layer on said surface portion; and
(c) a polymer layer formed on said linking layer by a process of surface-initiated polymerization of monomeric units thereon, 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 head group projecting from said stem.

2. The biomolecular detector of claim 1 , further comprising:
(d) a first member of a specific binding pair coupled to said brush molecule.

3. The detector of claim 2 , wherein said member of a specific binding pair further comprises an extended nucleic acid conjugated thereto, said extended nucleic acid produced by a process of enzymatic extension with terminal transferase.

4. The detector of claim 2 , wherein said first member is a first nucleic acid, said detector optionally further comprising a second nucleic acid as a second member of said binding pair hybridized to said first member.

5. The detector of claim 4 , further comprising an extended nucleic acid produced by a process of enzymatic extension with terminal transferase coupled to either said first or second nucleic acid.

6. The detector of claim 2 , wherein said member of a specific binding pair is a protein or peptide.

7. The detector of claim 6 , further comprising: a second member of said specific binding pair coupled to said first member of said specific binding pair; an antibody specifically bound to said second member to form a sandwich, said antibody having a first nucleic acid conjugated thereto; and an extended nucleotide coupled to said first nucleotide, said extended nucleic acid produced by a process of enzymatic extension with terminal transferase.

8. The detector of claim 2 , wherein said first member is an antibody.

9. The detector of claim 1 , wherein said surface portion comprises a material selected from the group consisting of metals, metal oxides, semiconductors, polymers, silicon, silicon oxide, and composites thereof.

10. The detector of claim 1 , wherein said surface portion comprises gold.

11. The detector of claim 1 , wherein said linking layer is continuous.

12. The detector of claim 1 , wherein said linking layer is patterned.

13. The detector of claim 1 , wherein said linking layer is a self-assembled monolayer.

14. The detector of claim 1 , wherein said brush molecule is from 5 to 50 nanometers in length.

15. The detector of claim 1 , wherein said brush molecule is formed on said surface portion at a density from 40 to 100 milligrams per meter 2 .

16. The detector of claim 1 , wherein said brush molecule is produced by surface-initiated atom transfer radical polymerization of oligo(ethylene glycol)methyl methacrylate.

17. The detector of claim 1 , wherein said head group comprises a kosmotrope.

18. The detector of claim 2 , wherein said first member of said specific binding pair is coupled directly to a hydroxyl group in OEGMA by CDI chemistry.

Assignees

• Univ Duke

Inventors

• Chilkoti Ashutosh

Classifications

• B01J19/0046 Primary

  Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays (synthesis methods per se C40B50/00) · CPC title

• B01J2219/00612

  the surface being inorganic · CPC title

• G01N33/5302

  Apparatus specially adapted for immunological test procedures · CPC title

• B01J2219/00637

  by coating it with another layer · CPC title

• B01J2219/0074

  Biological products · CPC title