Use of microfluidic systems in the electrochemical detection of target analytes

• US 9,151,746 B2
• Filed: 06/18/2013
• Issued: 10/06/2015
• Est. Priority Date: 04/21/1999
• Status: Expired due to Fees

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: Osmetech Technology Inc. (Roche Holding AG)
• Inventors: Kayyem, Jon Faiz
• Primary Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US13/921,117
• Publication Number:
  US 20140087374A1
• Time in Patent Office: 840 Days
• Field of Search: 20440301-40315, 204/411, 204/412, 204/452, 204/603, 205/777.5, 205/778, 205/792
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/021 Identification, e.g. bar codes
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0418 electro-osmotic flow [EOF]
• CPC Class Codes: B01L 2400/0439 ultrasonic vibrations, vibrating piezo elements
• CPC Class Codes: B01L 2400/0442 thermal energy, e.g. vaporisation, bubble jet
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0611 duck bill valves
• CPC Class Codes: B01L 2400/0633 with moving parts
• CPC Class Codes: B01L 2400/0638 membrane valves, flap valves
• CPC Class Codes: B01L 2400/0677 phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502738 : characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502761 : specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 7/525 : with physical movement of samples between temperature zones
• CPC Class Codes: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: B82Y 35/00 : Methods or apparatus for measurement or analysis of nanostructures
• CPC Class Codes: B82Y 40/00 : Manufacture or treatment of nanostructures
• CPC Class Codes: C12Q 1/6813 : Hybridisation assays
• CPC Class Codes: C12Q 1/6825 : Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2563/113 : the label being electroactive, e.g. redox labels
• CPC Class Codes: C12Q 2565/629 : being a microfluidic device
• CPC Class Codes: G01N 2035/00158 : Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 2610/00 : Assays involving self-assembled monolayers [SAMs]
• CPC Class Codes: G01N 27/3275 : Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
• CPC Class Codes: G01N 27/3276 : being a hybridisation with immobilised receptors using a FET type sensor G01N27/4145; concerning the hybridisation C12Q1/68
• CPC Class Codes: G01N 27/447 : using electrophoresis
• CPC Class Codes: G01N 27/4473 : by electric means
• CPC Class Codes: G01N 27/44752 : Controlling the zeta potential, e.g. by wall coatings
• CPC Class Codes: G01N 27/44791 : Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 33/5302 : Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/5438 : Electrodes
• CPC Class Codes: G01N 33/54386 : Analytical elements

Abstract

The invention relates generally to methods and apparatus for conducting analyses, particularly microfluidic devices for the detection of target analytes.

Claims

1. A microfluidic device for the detection of a target analyte in a fluid sample, said device comprising a solid support member comprising:

a) a sample inlet port;

b) a configuration of electrodes for electrokinetic movement of said sample on said solid support member upon application of a voltage; and

c) a detection chamber comprising a plurality of detection electrodes positioned in said detection chamber, each comprising a binding ligand and a self assembled monolayer, wherein said detection chamber is in fluid connection to said sample inlet port;

wherein said electrokinetic movement is electroosmotic movement or electrohydrodynamic movement.

2. The microfluidic device according to claim 1 wherein said electrokinetic movement is electroosmotic movement.

3. The microfluidic device according to claim 2 wherein said binding ligand is a nucleic acid.

4. The microfluidic device according to claim 2 wherein said binding ligand is a protein.

5. The microfluidic device according to claim 1 wherein said electrokinetic movement is electrohydrodynamic movement.

6. A method for the detection of a target analyte in a sample, said method comprising:

a) introducing said sample to a sample inlet port of a microfluidic device according to claim 1;

b) applying a voltage to said configuration of electrodes to move said sample to said detection chamber;

c) forming an attachment complex between said target analyte and at least one of said binding ligands, wherein said attachment complex comprises an electron transfer moiety label; and

d) detecting the presence of said label as an indication of the presence of said target.

7. A method according to claim 6 wherein said electron transfer moiety label is a metallocene.

8. A method according to claim 7 wherein said metallocene is a ferrocene.

9. A method according to claim 6 wherein said attachment complex comprises said target analyte, said capture binding ligand, and a solution binding ligand comprising said electron transfer moiety label.

10. A microfluidic device for the detection of a target analyte in a fluid sample, said device comprising a solid support member comprising:

a) a sample inlet port;

b) a configuration of electrodes for an electrokinetic movement of said sample on said solid support member upon application of a voltage;

c) a detection chamber comprising a plurality of detection electrodes positioned in said detection chamber, each comprising a binding ligand and a self-assembled monolayer, wherein said detection chamber is in fluid connection to said sample inlet port;

wherein said binding ligand is a nucleic acid or a protein.

11. The microfluidic device of claim 10, wherein said electrokinetic movement is electroosmotic movement.

12. The microfluidic device of claim 10, wherein said electrokinetic movement is electrohydrodynamic movement.

13. The microfluidic device of claim 10, wherein said binding ligand is a nucleic acid.

14. The microfluidic device of claim 10, wherein said binding ligand is a protein.

15. A method for the detection of a target analyte in a sample, said method comprising:

a) introducing said sample to a sample inlet port of a microfluidic device according to claim 10;

b) applying a voltage to said configuration of electrodes to move said sample to said detection chamber;

c) forming an attachment complex between said target analyte and at least one of said binding ligands, wherein said attachment complex comprises an electron transfer moiety label; and

d) detecting the presence of said label as an indication of the presence of said target.

16. The method of claim 15, wherein said electron transfer moiety label is a metallocene.

17. The method of claim 16, wherein said metallocene is a ferrocene.

18. The method of claim 15, wherein said attachment complex comprises said target analyte, said capture binding ligand, and a solution binding ligand comprising said electron transfer moiety label.