Conductive microplate

• US 20030215937A1
• Filed: 05/15/2002
• Published: 11/20/2003
• Est. Priority Date: 05/15/2002
• Status: Active Grant

• Current Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Original Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Inventors: Matson, Robert S.
• Granted Patent: US 6,841,379 B2
• Time in Patent Office: Days
• US Class Current: 435/287.2
• 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 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00639 the compounds being trapped in or bound to a porous medium
• 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 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2300/069 Absorbents; Gels to retain a fluid
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

A conductive microplate device for the detection of target biomolecules in a sample is described. The microplate comprises an assembly of a porous substrate and a conductive layer, wherein the assembly is sealed into bottom of at least some wells of the microplate. The porous substrate has a top surface and a bottom surface. The top surface comprises a plurality of covalently attached probe biomolecules. The covalently attached probe biomolecules are reactive with the target biomolecules contained in the sample. The conductive layer, which is attached to the bottom surface of the porous substrate, is adapted to receive voltage. Microplates of the present invention can be easily adapted for use with robotic workstations. Accordingly, in one embodiment, the power supply is incorporated into a robotic arm tool for fast microplate processing.

Claims

1. A conductive microplate for detecting target biomolecules in a sample, the microplate with a plurality of wells, the microplate comprising:

an assembly of a porous substrate and a conductive layer, wherein the assembly is sealed into bottom of at least some wells,

wherein

the porous substrate has a top surface and a bottom surface,

the top surface comprises a plurality of covalently attached probe biomolecules, wherein the probe biomolecules are reactive with target biomolecules,

the conductive layer is attached to the bottom surface of the porous substrate,

and the conductive layer is adapted to receive voltage.

2. The microplate of claim 1, wherein the top surface further comprises a plurality of activated pendant functional groups reactive with probe biomolecules;

and the top surface further comprises a plurality of probe biomolecules covalently bound to the pendant functional groups.

3. The microplate of claim 2, wherein the activated pendant functional groups are acyl fluoride groups.

4. The microplate of claim 3, wherein the probe biomolecules are attached to the top surface of the porous substrate without modification.

5. The microplate of claim 1, wherein the probe biomolecules are attached at discrete locations on the top surface of the porous substrate, whereby an array is formed.

6. The microplate of claim 5, wherein the array in each well comprises from about 100 to about 400 probe biomolecules.

7. The microplate of claim 1, wherein

the conductive layer forms a first electrode and the microplate further comprises:

a second electrode in an electrical contact with the first electrode.

8. The microplate of claim 7, wherein the electrical contact between the first and the second electrode is effectuated by an electrolyte solution.

9. The microplate of claim 1, wherein the conductive layer is attached directly to the bottom surface of the porous substrate.

10. The microplate of claim 9, wherein the conductive layer is a metallic coating.

11. The microplate of claim 1 further comprising a permeation layer disposed between the bottom surface of the porous substrate and the conductive layer.

12. The microplate of claim 11, wherein the permeation layer comprises a conductive material selected from a group consisting of agaroses and cross-linked polymers.

13. The microplate of claim 12, wherein the permeation layer comprises a methacrylate.

14. The microplate of claim 1, wherein the porous substrate is prepared from a material that comprises pendant carboxyl groups.

15. The microplate of claim 14, wherein the porous substrate is selected from a group consisting of carboxylated polypropylene, carboxylated polyethylene, and polycarbonates.

16. The microplate of claim 1, wherein the porous substrate is prepared from a material that comprises a coating having pendant carboxyl groups.

17. The microplate of claim 1, wherein the porous substrate has pores with a diameter of 16 to 18 μ

m.

18. The microplate of claim 1, wherein the microplate comprises from 96 to 384 wells.

19. The microplate of claim 1, wherein the probe and the target biomolecules are selected from a group consisting of:

nucleic acids, polynucleotides, polypeptides, proteins, carbohydrates, lipids, and analogs thereof.

20. The microplate of claim 19, wherein the biomolecule is a polynucleotide selected from a group consisting of amplified DNA, cDNA, single-stranded DNA, double-stranded DNA, PNA, RNA, or mRNA.

21. A device comprising the microplate of claim 7 and a power supply for supplying voltage to the first and the second electrodes.

22. The device of claim 21, wherein the power supply is a hot plate comprising:

a surface adapted to accept the microplate;

a power source; and

means for supplying voltage from the power source to the first and the second electrodes of the microplate.

23. The device of claim 22, wherein the means for supplying voltage comprise leads incorporated into the hot plate, wherein the leads mate with the first and the second electrodes of the microplate when the microplate is positioned on the surface of the hot plate.

24. The device of claim 22, wherein the first electrode comprises a network of electrode patches sealed into the base of individual wells and the hot plate comprises a matching network of electrical leads such that a voltage is transmitted from the electrical leads to electrode patches when the microplate is positioned on the surface of the hot plate.

25. The device of claim 21, wherein the first electrode comprises a network of electrode patches sealed into the base of individual wells and the power supply comprises a matching network of electrical leads such that when the microplate is assembled the electrode patches mate with the matching electrical leads.

26. A device comprising microplate of claim 1 and a robotic arm supplying voltage to the conductive layer, the device comprises an electrode prong tool having one electrode in an electrical contact with the conductive layer.

27. The device of claim 26, further comprising a second electrode in electrical contact with the first electrode through an electrolyte solution.

28. The device of claim 26, wherein the first electrode is pierced through the porous substrate.

33. A method for hybridizing target biomolecules to probe biomolecules, the method comprising:

(a) providing the conductive microplate of claim 8;

(b) introducing labeled target biomolecules into the electrolyte solution;

(c) applying an electrical voltage between the first and the second electrode, whereby the labeled target biomolecules hybridize with the probe biomolecules of the surface of the porous substrate.

29. A method of forming a conductive microplate having a plurality of wells, the method comprising:

(a) providing a porous substrate with a top surface and a bottom surface;

(b) activating the top surface with reactive functional groups capable of covalent attachment of probe biomolecules;

(c) contacting the top surface with probe biomolecules under conditions sufficient for covalent attachment of probe biomolecules to reactive functional groups;

(d) attaching a conductive layer to the bottom surface of the porous substrate; and

(e) sealing the porous substrate with the conductive layer into bottom of at least some wells of the microplate, wherein the conductive layer is adapted to receive voltage.

30. The method of claim 29, wherein the contacting step (c) comprises contacting the probe biomolecules at discrete locations on the porous substrate, whereby a bioarray is formed.

31. The method of claim 29, wherein the contacting step (c) is carried out by a technique selected from a group consisting of jet printing, solid or open capillary device contact printing, microfluidic channel printing, silk screening, and a technique using printing devices based upon electrochemical or electromagnetic forces.

32. The method of claim 29, wherein the attaching step (d) is carried out by a technique selected from a group consisting of silk screening, chemical deposition of metal, gluing, lamination, encapsulation, and polymerization.