Microvolume immunoabsorbant assays with amplified electrochemical detection

• US 20020058279A1
• Filed: 10/15/2001
• Published: 05/16/2002
• Est. Priority Date: 10/16/2000
• Status: Active Grant

• Current Assignee: University of Arkansas Board of Directors
• Original Assignee: University of Arkansas Board of Directors
• Inventors: Fritsch, Ingrid, Beittle, Robert Jr.
• Granted Patent: US 6,887,714 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01F 33/3032 using magneto-hydrodynamic [MHD] phenomena to mix or move the fluids
• CPC Class Codes: B01J 19/0046 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 Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00653 the compounds being bound to electrodes embedded in or on the solid supports
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00704 integrated with the reactor apparatus
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/043 magnetic forces
• 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 3/502761 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 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: F04B 17/00 Pumps characterised by combination with, or adaptation to, specific driving engines or motors
• CPC Class Codes: G01N 27/3277 being a redox reaction, e.g. detection by cyclic voltammetry voltammetry per se G01N27/42, G01N27/48
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

A structure and method for chemical sensing utilizing microassays. Microcavities or micropores are combined with assay techniques to provide a very fast and very sensitive means of detecting chemical compounds. Assay techniques are modified to include a metal ion binding carrier species especially suitable for use in conjunction with the electrochemical detection. This allows assays to be combined with electrochemical analysis, thus allowing the high speed ease and hypersensitivity available in the invention disclosed herein.

Claims

1. A microassay structure comprising:

a microcavity formed of insulating layers and integrated, independently addressable electrodes; and

,

an analyte binding material.

2. The microassay structure of claim 1 wherein said analyte binding material is selected from the group consisting of primary antibodies, polynucleotides, lipid layers, or a protein binding compound.

3. The microassay structure of claim 1 wherein said microcavity is less than 70 micrometers in diameter.

12. The microassay structure of claim 1 further comprising a lipid bilayer suspended across the opening of said microcavity.

16. The method of claim 13 wherein said carrier species is a synthetic dendromer.

An array of microassay structures comprising:

At least one of the microstructures according to claim 1.

4. A microassay structure comprising:

a micropore formed of insulating layers and integrated, independently addressable electrodes; and

,

an analyte binding material.

5. The microassay structure of claim 4 wherein said analyte binding material is selected from the group consisting of primary antibodies, polynucleotides, lipid layers or a protein binding material.

6. The microassay structure of claim 4 wherein said pore is less than 100 micrometers in diameter.

17. An array of microassay structures comprising:

at least one of the microassay structures according to claim 4.

A method for detecting chemical compounds comprising;

addition of a sample to immunomicrobeads;

allowing an analyte in said sample to bind to an analyte binding material on the surface of said immuno microbeads;

rinsing said immunomicrobeads such that said analyte remains bound to said analyte binding material, while the remainder of said sample is removed;

incorporating said immunomicrobeads into a microassy structure;

addition of a secondary antibody having a carrier species attached;

allowing said secondary antibody to bind to said analyte;

activating said carrier species; and

,

measuring a current through said micro assay structure.

7. A method for detecting chemical compounds comprising:

addition of a sample to a microassay structure;

allowing an analyte in said sample to bind to a analyte binding material in said microimmunoassay structure;

rinsing said microimmunoassay structure such that said analyte remains bound to said analyte binding material while the remainder of said sample is removed;

addition of a secondary antibody having a carrier species attached;

allowing said secondary antibody to bind to said analyte;

activating said carrier species; and

,

measuring a current through said microassay structure.

8. The method of claim 7 wherein said carrier species is a metallo protein.

9. The method of claim 7 wherein said carrier species is a syntheticdendriner.

10. The method of claim 7 wherein said activation of said carrier species is selected from the group consisting of change in temperature, change in pH, application of electrical current, exposure to electromagnetic radiation or addition of an activating compound.

11. The method of claim 7 wherein said carrier species is selected from the group consisting of metallo protein, enzyme, dndrimer or kelating agents.

14. The method of claim 13 wherein said carrier species is a metallo protein.

15. The method of claim 13 wherein said activation of said carrier species is selected from the group consisting of change in temperature, change in pH, application of electrical current, exposure to electromagnetic radiation or addition of an activating compound.

18. The method of claim 19 wherein said carrier species is a metallo protein.

19. The method of claim 19 wherein said carrier species is a synthetic dendrimer.

20. The method of claim 19 wherein said activation of said carrier species is selected from the group consisting of change in temperature, change in pH, application of electrical current, exposure to electromagnetic radiation or addition of an activating compound.

13. A method for detecting chemical compounds comprising:

addition of a sample to a microassay structure;

allowing an analyte in said sample to bind to a analyte binding material in said microassay structure;

rinsing said microassay structure such that said analyte remained bound to said analyte binding material while the remainder of said sample is removed;

addition of a second analyte having a carrier species attached;

allowing said secondary antibody to bind to said analyte;

activating said carrier species; and

,

measuring a current through said microassay structure.