Enzymatic signal generation and detection of binding complexes in stationary fluidic chip

• US 20080160630A1
• Filed: 12/29/2006
• Published: 07/03/2008
• Est. Priority Date: 12/29/2006
• Status: Active Grant

• Current Assignee: Intel Corporation
• Original Assignee: Intel Corporation
• Inventors: Wu, Kai, Su, Xing, Liu, David J.
• Granted Patent: US 8,409,877 B2
• Time in Patent Office: Days
• US Class Current: 436/164
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• 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 3/502761 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: B03C 1/01 by addition of magnetic adjuvants
• CPC Class Codes: B03C 1/288 disposed at the outer circumference of a recipient
• CPC Class Codes: B03C 1/30 Combinations with other devices, not otherwise provided for
• CPC Class Codes: B03C 2201/18 Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: B03C 2201/26 for use in medical applications
• 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 21/255 Details, e.g. use of specially adapted sources, lighting or optical systems
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 21/658 enhancement Raman, e.g. surface plasmons
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 33/54333 Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction
• CPC Class Codes: G01N 33/587 Nanoparticles
• CPC Class Codes: Y10T 436/25 including sample preparation

Abstract

An embodiment of the invention relates to a device for detecting an analyte in a sample. The device comprises a fluidic network and an integrated circuitry component. The fluidic network comprises a sample zone, a cleaning zone and a detection zone. The fluidic network contains a magnetic particle and/or a signal particle. A sample containing an analyte is introduced, and the analyte interacts with the magnetic particle and/or the signal particle through affinity agents. A microcoil array or a mechanically movable permanent magnet is functionally coupled to the fluidic network, which are activatable to generate a magnetic field within a portion of the fluidic network, and move the magnetic particle from the sample zone to the detection zone. A detection element is present which detects optical or electrical signals from the signal particle, thus indicating the presence of the analyte.

Claims

1. A method of detecting an analyte, comprising

providing a magnetic affinity complex and a signal affinity complex in a fluidic network comprising a plurality of fluidic zones, wherein the plurality of fluidic zones comprises a sample zone, a cleaning zone, and a detection zone, wherein the fluidic network is functionally coupled to a vibration element, and wherein the signal affinity complex comprises a SERS-active nanoparticle, a fluorescent nanoparticle, an optical nanoparticle, a MRI-active nanoparticle, and optionally comprises a catalytic element,

introducing a sample suspected of comprising an analyte into the sample zone, wherein the analyte combines with the magnetic affinity complex and the signal affinity complex to form a sandwich binding complex,

activating a microcoil array or a mechanically movable permanent magnet functionally coupled to the fluidic network to thereby move the sandwich binding complex to the detection zone without fluidic movement of a fluid in the plurality of the fluidic zones, and

detecting the presence of the sandwich binding complex within the detection zone using a detection element functionally coupled to the fluidic network.

2. The method of claim 1, wherein the detection element is an optical detection element or an electrical detection element.

3. The method of claim 1, wherein the signal affinity complex comprises a SERS-active nanoparticle, an optical nanoparticle, or a fluorescent nanoparticle.

4. The method of claim 1, wherein the signal affinity complex comprises a catalytic element.

5. The method of claim 4, wherein the catalytic element is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, glucose oxidase, glucose oxidase and horseradish peroxidase, firefly luciferase, Renilla luciferase, bacterial luciferase, an enzyme or analogs or combinations thereof.

6. The method of claim 4, wherein the catalytic element is conjugated through a functionalized polymer.

7. The method of claim 4, wherein a reaction substrate for the catalytic element is in the detection zone.

8. The method of claim 7, wherein the reaction substrate is selected from the group consisting of Lumi-Phos 480, Lumi-Phos 530, Lumi-Phos Plus, Lumi-Phos APS-5, Lumigen TMA-6, Lumigen PS-atto, Lumigen PS-1, Lumigen PS-2, Lumigen PS-3, H₂O₂ with an oxidizable compound, Lumi-Gal 530, Amplex Red, 3,5,3′

,5′

-tetramethylbenzidine (TMB), glucose, O₂, ATP, Mg²⁺, luciferin, aminoluciferin, quinolinyl lucifiern, coelentrazine, aldehyde, FMNH₂, and analogs and derivatives, and combinations thereof.

9. The method of claim 4, wherein the fluidic zones contain an appropriate buffer.

10. The method of claim 1, wherein before the sandwich binding complex reaches the detection zone, it is moved to the cleaning zone, wherein the vibration element is activated to aggregate and de-aggregate the binding complex to thereby remove unbound signal affinity complex.

11. A method of detecting an analyte, comprising

providing a magnetic affinity complex and a signal analyte complex in a fluidic network comprising a plurality of fluidic zones, wherein the plurality of fluidic zones comprises a sample zone, a cleaning zone, and a detection zone, wherein the fluidic network is functionally coupled to a vibration element, and wherein the signal analyte complex comprises a SERS-active nanoparticle, an optical nanoparticle, a fluorescent nanoparticle, and optionally comprises a catalytic element,

introducing a sample suspected of comprising an analyte into the sample zone, wherein the analyte combines with the magnetic affinity complex to form a magnetic binding complex,

activating a microcoil array or a mechanically movable permanent magnet functionally coupled to the fluidic network to thereby move the magnetic binding complex within the fluidic zone or to another fluidic zone without fluidic movement of a fluid in the plurality of the fluidic zones,

displacing the analyte from the magnetic binding complex with the signal analyte complex to form a competitive binding complex,

optionally activating the array of microcoils or mechanically movable permanent magnet to move the competitive binding complex within the fluidic network prior to detecting the competitive binding complex and/or the unbound signal analyte complex, and

detecting the presence of the unbound signal analyte complex within the detection zone using a detection element functionally coupled to the fluidic network, wherein the detection element is a optical detection element or an electrical detection element.

12. The method of claim 11, wherein the signal analyte complex comprises a SERS-active nanoparticle, an optical nanoparticle, or a fluorescent nanoparticle.

13. The method of claim 11, wherein the signal analyte complex comprises a catalytic element.

14. The method of claim 13, wherein the catalytic element is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, glucose oxidase, glucose oxidase and horseradish peroxidase, firefly luciferase, Renilla luciferase, bacterial luciferase, an enzyme or analogs or combinations thereof.

15. The method of claim 13, wherein the catalytic element is conjugated through a functionalized polymer.

16. The method of claim 13, wherein a reaction substrate for the catalytic element is in the detection zone.

17. The method of claim 16, wherein the reaction substrate is selected from the group consisting of Lumi-Phos 480, Lumi-Phos 530, Lumi-Phos Plus, Lumi-Phos APS-5, Lumigen TMA-6, Lumigen PS-atto, Lumigen PS-1, Lumigen PS-2, Lumigen PS-3, H₂O₂ with an oxidizable compound, Lumi-Gal 530, Amplex Red, 3,5,3′

,5′

-tetramethylbenzidine (TMB), glucose, O₂, ATP, Mg²⁺, luciferin, aminoluciferin, quinolinyl lucifiem, coelentrazine, aldehyde, FMNH₂, and analogs and derivatives, and combinations thereof.

18. The method of claim 13, wherein the fluidic zones contain an appropriate buffer.

19. A method of detecting an analyte, comprising

providing a coded magnetic affinity complex and a magnetic signal affinity complex in a fluidic network comprising a plurality of fluidic zones, wherein the plurality of fluidic zones comprises a sample zone, a cleaning zone, and a detection zone, wherein the fluidic network is functionally coupled to a vibration element, and wherein the magnetic signal affinity complex comprises a SERS-active nanoparticle, a fluorescent nanoparticle, an optical nanoparticle, a MRI-active nanoparticle, and optionally comprises a catalytic element,

introducing a sample suspected of comprising an analyte into the sample zone, wherein the analyte combines with the coded magnetic affinity complex to form a coded magnetic binding complex,

activating a microcoil array or a mechanically movable permanent magnet functionally coupled to the fluidic network to thereby move the coded magnetic binding complex to a first affinity surface without fluidic movement of a fluid in the plurality of the fluidic zones, forming a bound coded magnetic binding complex,

detaching the code from the bound coded magnetic binding complex,

providing a magnetic signal affinity complex, wherein the detached code binds to the magnetic signal particle to form a coded magnetic signal binding complex,

activating the microcoil array or mechanically movable permanent magnet to move the coded signal binding complex to the detection zone comprising a second affinity surface, forming a bound coded signal binding complex, and

detecting the bound coded signal binding complex within the detection zone using a detection element functionally coupled to the fluidic network.

20. The method of claim 19, wherein the magnetic signal affinity complex comprises a SERS-active nanoparticle or a fluorescent nanoparticle.

21. The method of claim 19, wherein the magnetic signal affinity complex comprises a catalytic element.

22. The method of claim 21, wherein the catalytic element is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, glucose oxidase, glucose oxidase and horseradish peroxidase, firefly luciferase, Renilla luciferase, bacterial luciferase, an enzyme or analogs or combinations thereof.

23. The method of claim 21, wherein the catalytic element is conjugated through a functionalized polymer.

24. The method of claim 21, wherein a reaction substrate for the catalytic element is in the detection zone.

25. The method of claim 24, wherein the reaction substrate is selected from the group consisting of Lumi-Phos 480, Lumi-Phos 530, Lumi-Phos Plus, Lumi-Phos APS-5, Lumigen TMA-6, Lumigen PS-atto, Lumigen PS-1, Lumigen PS-2, Lumigen PS-3, H₂O₂ with an oxidizable compound, Lumi-Gal 530, Amplex Red, 3,5,3′

,5′

-tetramethylbenzidine (TMB), glucose, O₂, ATP, Mg²⁺, luciferin, aminoluciferin, quinolinyl lucifiern, coelentrazine, aldehyde, FMNH₂, and analogs and derivatives, and combinations thereof.

26. The method of claim 21, wherein the fluidic zones contain an appropriate buffer.

27. The method of claim 19, wherein the coded magnetic binding complex and/or the coded signal binding complex are moved to a cleaning zone by activating the microcoil array or mechanically moveable permanent magnet, wherein the vibration element is activated to aggregate and de-aggregate the complex to thereby remove unbound coded magnetic affinity complex, detached code, and/or magnetic signal affinity complex.

28. The method of claim 19, wherein the detection element is an optical detection element or an electrical detection element.

29. A method of detecting an analyte, comprising

providing a magnetic signal affinity complex in a fluidic network comprising a plurality of fluidic zones, wherein the plurality of fluidic zones comprises a sample zone, a cleaning zone, and a detection zone, wherein the fluidic network is functionally coupled to a vibration element, and wherein the magnetic signal affinity complex comprises a SERS-active nanoparticle, a fluorescent nanoparticle, and/or comprises a catalytic element,

introducing a sample suspected of an analyte into the sample zone, wherein the analyte combines with the magnetic signal affinity complex to form a magnetic signal binding complex,

activating a microcoil array or a mechanically movable permanent magnet functionally coupled to the fluidic network to thereby move the magnetic signal binding complex to the detection zone without fluidic movement of a fluid in the plurality of the fluidic zones,

detecting the presence of the binding complex within the detection zone using a detection element functionally coupled to the fluidic network, wherein the detection element is a optical detection element or an electrical detection element.