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

Patent metadata

Field	Value
Publication number	US-11130989-B2
Application number	US-201815872392-A
Country	US
Kind code	B2
Filing date	Jan 16, 2018
Priority date	Sep 15, 2005
Publication date	Sep 28, 2021
Grant date	Sep 28, 2021

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 system for detecting the presence of a target molecule in a sample, the system comprising:
(a) a capture component, the capture component comprising: (i) a substrate having a surface portion; (ii) a linking layer on said surface portion; (iii) a polymer layer formed on said linking layer by the 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; and (iv) a first member of a specific binding pair coupled to said brush molecule;
(b) a signal source that emits an incident signal that interrogates the capture component after the sample has been applied to the capture component;
(c) a signal amplifier, wherein a post-incident signal that is generated after the incident signal interrogates the capture component is amplified to increase the signal-to-noise ratio when the target molecule is bound to the capture component; and
(d) a signal detector for detecting the amplified post-incident signal.

2. The system of claim 1 , wherein the target molecule to be detected comprises DNA, small molecules, peptides, oligosaccharides and carbohydrates, wherein the incident signal is light, plasmons or ions.

3. The system of claim 2 , wherein the post-incident signal is detected by: (a) time-of-flight secondary-ion mass spectroscopy (ToF-SIMS), (b) matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), (c) surface plasmon resonance (SPR) spectroscopy, (d) quantum dots or (e) metal nanoparticles.

4. The system of claim 2 , wherein the target molecule is DNA.

5. The system of claim 4 , wherein the first member of the specific binding pair is a single-stranded DNA molecule that is coupled the brush molecule via the 3′ terminus of the single-stranded DNA.

6. The system of claim 5 , wherein the brush molecule is poly(MAA-co-OEGMA).

7. The system of claim 6 , wherein one strand of the DNA target molecule can hybridize to the single-stranded DNA molecule coupled to the brush molecule.

8. The system of claim 7 , wherein the target DNA that hybridizes to the single-stranded DNA molecule coupled to the brush molecule can be extended at its 3′ terminus.

9. The system of claim 5 , wherein the first member of the specific binding pair is a single-stranded DNA molecular beacon that is coupled the brush molecule via the 5′ terminus of the single-stranded DNA molecular beacon.

10. The system of claim 9 , wherein the binding of the target DNA to the molecular beacon causes unfolding of the molecular beacon such that the 3′ terminus of the molecular beacon becomes accessible for extension by terminal transferase (TdTase).

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

• B01J2219/00725

  Peptides · CPC title

• B01J2219/0063

  Other, e.g. van der Waals forces, hydrogen bonding · CPC title

• B01J2219/00637

  by coating it with another layer · CPC title

• C40B50/18

  using a particular method of attachment to the solid support · CPC title