Self-contained microelectrochemical bioassy platforms and methods

• US 20030077642A1
• Filed: 09/24/2002
• Published: 04/24/2003
• Est. Priority Date: 10/16/2000
• Status: Active Grant

• Current Assignee: Board of Trustees of The University of Arkansas NA
• Original Assignee: Board of Trustees of The University of Arkansas NA
• Inventors: Aguilar, Zoraida, Beitle, Robert Jr., Fritsch, Ingrid
• Granted Patent: US 7,348,183 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 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• 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 27/3278 involving nanosized elements, e.g. nanogaps or nanoparticles nanopores G01N33/48721; magnetic beads G01N27/745
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

Methods and devices for improved chemical and biomass detection assays combined well defined microstructures having independently addressable electrodes with various surface immobilization electrochemical assays. Combining known chemical detection immobilization assays, electrochemically active moieties with microstructures having independently addressable electrodes provides for vastly improved methods of detecting microorganisms, chemical compounds, and measuring membrane transport.

Claims

1. A microassay structure comprising:

at least one microstructure 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 microstructure is less than 100 micrometers in diameter.

4. The microassay structure of claim 1 wherein said microstructure is a microcavity.

5. The microassay structure of claim 1 wherein said microstructure is a micropore.

6. The microassay structure of claim 1 wherein said analyte binding material is comprised of primary antibodies covalently bound to a self assembled monlayer.

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

14. A microassay structure of claim 1 comprising a plurality of microstructures.

7. A method for detecting an analyte comprising:

addition of a sample to a microassay structure;

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

addition of a secondary antibody having an electroactive complex attached;

allowing said secondary antibody to bind to said analyte;

activating said electroactive complex; and

,

measuring electric current generated by the electroactive complex.

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

9. The method of claim 7 wherein said electroactive complex is selected from the group consisting of a metalloprotein, redox enzyme, dendrimer or chelating agents.

11. A method for detecting a polynucleotide sequence comprising:

addition of a sample to a microassay structure, wherein said microassay structure is comprised of at least one analyte binding material and at least one electrode;

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

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

addition of a polynucleotide probe having a covalently bound electroactive complex;

allowing said polynucleotide probe to bind to said polynucleotides;

activating said electroactive complex; and

,

measuring a current generated by electroactive complex.

12. The method of claim 11 wherein said carrier species is selected from the group consisting of metalloproteins, dendrimers or redox enzymes.

13. The method of claim 11 wherein said activation of said electroactive complex 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.

15. 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 microassay structure;

addition of a secondary antibody having an electroactive complex attached;

allowing said secondary antibody to bind to said analyte;

rinsing said microassay structure such that only said secondary antibodies bound to said analyte remain in said microassay structures;

activating said electroactive complex; and

,

measuring a current through said micro assay structure.

16. The method of claim 15 wherein said electroactive complex is selected from the group consisting of metalloproteins, dendrimers or redox enzymes.

17. The method of claim 15 wherein said activation of said electroactive complex 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.