Nano-mechanic microsensors and methods for detecting target analytes

• US 7,473,551 B2
• Filed: 05/20/2005
• Issued: 01/06/2009
• Est. Priority Date: 05/21/2004
• Status: Expired due to Fees

• Current Assignee: Atonomics A/S
• Original Assignee: Atonomics A/S
• Inventors: Warthoe, Peter
• Primary Examiner(s): Chin; Christopher L
• Application Number:
  US11/134,821
• Publication Number:
  US 20060024813A1
• Time in Patent Office: 1,327 Days
• Field of Search: None
• US Class Current: 435/287.2
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 2291/0255 (Bio)chemical reactions, e.g. on biosensors
• CPC Class Codes: G01N 2291/0256 Adsorption, desorption, surface mass change, e.g. on biosensors
• CPC Class Codes: G01N 2291/0422 Shear waves, transverse waves, horizontally polarised waves
• CPC Class Codes: G01N 2291/0423 Surface waves, e.g. Rayleigh waves, Love waves
• CPC Class Codes: G01N 2291/0427 Flexural waves, plate waves, e.g. Lamb waves, tuning fork, cantilever
• CPC Class Codes: G01N 29/022 Fluid sensors based on microsensors, e.g. quartz crystal-microbalance [QCM], surface acoustic wave [SAW] devices, tuning forks, cantilevers, flexural plate wave [FPW] devices microdevices per se B81B
• CPC Class Codes: G01N 29/226 Handheld or portable devices
• CPC Class Codes: G01N 29/2462 Probes with waveguides, e.g. SAW devices
• CPC Class Codes: G01N 29/30 Arrangements for calibrating or comparing, e.g. with standard objects

Abstract

The present invention relates generally to methods and compositions for analyzing test samples containing target analytes including proteins and nucleic acids. The invention uses a surface acoustic wave sensor in combination with a hydrogel to obtain an ultra sensitive non-fluorescent detection system.

Claims

1. A microsensor for detecting the presence of a target analyte in a test sample solution comprising a surface acoustic wave sensor comprising a hydrogel located on a surface of said sensor and at least one molecular recognition component immobilized within said hydrogel that is capable of detecting said target analyte, wherein the hydrogel comprises an array of charged colloidal particles within the hydrogel.

2. The microsensor according to claim 1, wherein a dimensional and or volume change of said hydrogel occurs when the target analyte binds to the molecular recognition component.

3. The microsensor according to claim 1, wherein two different molecular recognition components are capable of binding a single target analyte.

4. The microsensor according to claim 1, wherein the hydrogel comprises vinyl acetates, vinyl pyrrolidones, vinyl ethers, olefins, styrenes, vinyl chlorides, ethylenes, acrylates, methacrylates, nitriles, acrylamides, maleates, epoxies, epoxides, lactones, ethylene oxides, ethylene glycols, ethyloxazolines, amino acids, saccharides, proteins, anhydrides;

amides, carbonates, phenylene oxides, acetals, sulfones, phenylene sulfides, esters, fluoropolymers, imides, amide-imides, etherimides, ionomers, aryletherketones, amines, phenols, acids, benzenes, cinnamates, azoles, silanes, chlorides, and epoxides, N, N′

-methylenebisacrylamide, methylenebismethacrylamide ethyleneglycol- dimethacrylate, N, N′

-methylenebisacrylamide, polyethyleneglycoldiacrylate(PEGDA), polyethyleneglycoldimethacrylate(PEGDMA), polyethyleneglycoldiacrylate(PEGDA), polyethyleneglycoldimethacrylate(PEGDMA), poly(vinyliden fluoride)(PVdF)based polymer, a polyacrylonitrile(PAN) based polymer, a polymethylmethacrylate(PMMA) based polymer, a polyvinyl chloride(PVC) based polymer, and a mixture of the poly(vinyliden fluoride)(PVdF) based polymer, polyacrylonitrile(PAN) based polymer, polymethylmethacrylate(PMMA) based polymer, and polyvinyl hloride(PVC) based polymer, or mixtures thereof.

5. The microsensor according to claim 4, further comprising a cross linking agent wherein the cross linking agent comprises N,N′

- methylenebisacrylamide, methylenebismethacrylamide ethyleneglycol-dimethacrylate or agent N, N′

- methylenebisacrylamide.

6. The microsensor according to claim 1, wherein said surface acoustic wave sensor is a surface acoustic wave resonator.

7. The microsensor according to claim 1 , wherein the charged colloidal particles comprise polystyrene, polymethylmethacrylate, silicon dioxide, aluminum oxide, polytetrafluoroethylene or poly N-isopropylacrylamide.

8. The microsensor according to claim 1, further comprising a reference surface acoustic wave sensor comprising a reference hydrogel located on a surface of said reference sensor, wherein said reference hydrogel does not contain said molecular recognition component and wherein said microsensor is capable of subtracting the signal of said reference sensor from the signal of said sensor.

9. The microsensor according to claim 1, wherein the molecular recognition component is selected from the group consisting of nucleic acids, nucleotide, nucleoside, nucleic acids analogues such as PNA and LNA molecules, proteins, peptides, antibodies including IgA, IgG, IgM, IgE, enzymes, enzymes cofactors, enzyme substrates, enzymes inhibitors, membrane receptors, kinases, Protein A, Poly U, Poly A, Poly lysine, triazine dye, boronic acid, thiol, heparin, membrane receptors, polysaccharides, coomassie blue, azure A, metal-binding peptides, sugar, carbohydrate, chelating agents, prokaryotic cells or eukaryotic cells.

10. The microsensor according to claim 1, wherein the hydrogel comprises a polymerized monomer or hydrogel, a cross linking agent and optionally a chemical or UV-light activated inducer agent.

11. The microsensor according to claim 1, wherein the hydrogel comprises acrylamide, purified agarose, N-vinylpyrolidone, methacrylate, N-isopropylacrylamide, substituted acrylamide or poly(ethylene glycol) diacrylate (PEG-DA).

13. A handheld device for detecting target analytes comprising the microsensor of claim 1 or 12.

14. The microsensor of claim 13 wherein said channels have a height of 10 nm to 1 micron.

15. The microsensor of claim 14 wherein said channels have a width of 100 nm to 10 microns.

16. The microsensor according to claim 13, wherein a dimensional and or volume change of said hydrogel occurs when the target analyte binds to the molecular recognition component.

17. The microsensor according to claim 13, wherein two different molecular recognition components are capable of binding a single target analyte.

18. The microsensor according to claim 13, wherein the hydrogel comprises vinyl acetates, vinyl pyrrolidones, vinyl ethers, olefins, styrenes, vinyl chlorides, ethylenes, acrylates, methacrylates, nitriles, acrylamides, maleates, epoxies, epoxides, lactones, ethylene oxides, ethylene glycols, ethyloxazolines, amino acids, saccharides, proteins, anhydrides, amides, carbonates, phenylene oxides, acetals, sulfones, phenylene sulfides, esters, fluoropolymers, imides, amide-imides, etherimides, ionomers, aryletherketones, amines, phenols, acids, benzenes, cinnamates, azoles, silanes, chlorides, and epoxides, N, N′

-methylenebisacrylamide, methylenebismethacrylamide ethyleneglycol-dimethacrylate, N, N′

-methylenebisacrylamide, polyethyleneglycoldiacrylate(PEGDA), polyethyleneg lycoldimethacrylate(PEGDMA), polyethyleneglycoldiacrylate(PEGDA), polyethyleneglycoldimethacrylate(PEGDMA), poly(vinyliden fluoride)(PVdF) based polymer, a polyacrylonitrile(PAN) based polymer, a polymethylmethacrylate(PMMA) based polymer, a polyvinyl chloride(PVC) based polymer, and a mixture of the poly(vinyliden fluoride)(PVdF) based polymer, polyacrylonitrile(PAN) based polymer, polymethylmethacrylate(PMMA) based polymer, and polyvinyl hloride(PVC) based polymer;

or mixtures therof.

19. The microsensor according to claim 18, further comprising a cross linking agent wherein the cross. linking agent comprises N, N′

-methylenebisacrylamide, methylenebismethacrylamide ethyleneglycol-dimethacrylate or agent N, N′

-methylenebisacrylamide.

20. The microsensor according to claim 13, wherein said surface acoustic wave sensor is a surface acoustic wave resonator.

21. The microsensor of claim 13 wherein said hydrogel further comprises an array of charged colloidal particles within the hydrogel.

22. The microsensor according to claim 21 wherein the charged colloidal particles comprise polystyrene, polymethylmethacrylate, silicon dioxide, aluminum oxide, polytetrafluoroethylene or poly N-isopropylacrylamide.

23. The microsensor according to claim 13, further comprising a reference surface acoustic wave sensor comprising a reference hydrogel located on-a surface of said reference sensor, wherein said reference hydrogel does not contain said molecular recognition component and wherein said microsensor is capable of subtracting the signal of said reference sensor from the signal of said sensor.

24. The microsensor of claim 23 wherein the surface of said reference acoustic wave sensor comprises interdigitated electrodes having channels between said electrodes and wherein said channels contain all or part of said hydrogel.

25. The microsensor according to claim 13, wherein the molecular recognition component is selected from the group consisting of nucleic acids, nucleotide, nucleoside, nucleic acids analogues such as PNA and LNA molecules, proteins, peptides, antibodies including IgA, IgG, IgM, IgE, enzymes, enzymes cofactors, enzyme substrates, enzymes inhibitors, membrane receptors, kinases, Protein A, Poly U, Poly A, Poly lysine, triazine dye, boronic acid, thiol, heparin, membrane receptors, polysaccharides, coomassie blue, azure A, metal-binding peptides, sugar, carbohydrate, chelating agents, prokaryotic cells or eukaryotic cells.

26. The microsensor according to claim 13, wherein the hydrogel comprises a polymerized monomer or hydrogel, a cross linking agent and optionally a chemical or UV-light activated inducer agent.

27. The microsensor according to claim 13, wherein the hydrogel comprises acrylamide, purified agarose, N-vinylpyrolidone, methaaylate, N-isopropylacrylamide, substituted acrylamide or poly(ethylene glycol) diacrylate (PEG-DA).

12. A microsensor for detecting the presence of a target analyte in a test sample solution comprising a surface acoustic wave sensor comprising a hydrogel located on a surface of said sensor and at least one molecular recognition component immobilized within said hydrogel that is capable of detecting said target analyte, wherein said surface of said sensor comprises interdigitated electrodes having channels between said electrodes and wherein said channels contain all or part of said hydrogel.