Multiplex microarray of serially deposited biomolecules on a microarray

• US 20110281766A1
• Filed: 01/20/2011
• Published: 11/17/2011
• Est. Priority Date: 01/20/2010
• Status: Active Grant

• Current Assignee: CustomArray, Inc. (Genscript Biotech Corp.)
• Original Assignee: CombiMatrix Corporation, John Cooper
• Inventors: Cooper, John
• Granted Patent: US 9,927,434 B2
• Time in Patent Office: Days
• US Class Current: 506/16
• CPC Class Codes: G01N 33/5438 Electrodes

Abstract

Disclosed herein is a multiplex microarray having serially attached non-functionalized biomolecules attached to a polymer coating covering each electrode of an array of electrodes for assays and a method of making the multiplex microarray. The method comprises serially blocking the electrodes of the microarray with a blocking protein, electropolymerizing pyrrole or a functionalized pyrrole on the electrodes where the biomolecule is not present during polymerization, exposing the microarray to a biomolecular solution containing a non-functionalized biomolecule for attachment to the polymer coating, and then repeating the steps to form the multiplex microarray.

Claims

1. A method of making a multiplex microarray of serially attached different biomolecules on a microarray, comprising:

(a) blocking a microarray with a blocking protein, wherein the microarray is a device having an array of addressable microelectrodes on a surface of the device;

(b) electropolymerizing a monomer on one or more microelectrodes of the microarray to form a polymer coating on the one or more microelectrodes, wherein an electropolymerizing solution containing the monomer does not contain any of the biomolecules;

(c) exposing the microarray to a biomolecular solution containing a biomolecule for attachment to the polymer coating on the one or more electrodes, wherein the biomolecule is not an oligonucleotide and is not chemically functionalized;

(d) blocking the microarray with a blocking protein; and

(e) repeating steps (b) through (d) for each different biomolecule to be attached to the microarray, thereby making the microarray of different biomolecules.

2. The process of claim 1, wherein steps (a) and (d) further comprise:

washing the microarray to remove the blocking solution.

3. The process of claim 1, wherein step (b) further comprises:

washing the microarray to remove the electropolymerizing solution.

4. The process of claim 1, wherein step (c) further comprises:

washing the microarray to remove the biomolecular solution.

5. The process of claim 1, wherein the monomer is selected from the group consisting of pyrrole and functionalized pyrrole and combinations thereof.

6. The process of claim 1, wherein the monomer is selected from the group consisting of thiophenol, aniline, phenylene sulfide, monomers electropolymerizing to a conducting polymer, phenylenediamine, diaminohaphthalene, phenol, monomers electropolymerizing to a nonconducting polymer and phenolic derivatives, and combinations thereof.

7. The process of claim 1, wherein the biomolecules are selected from the group consisting of antibodies, antigens, proteins, Ricin, anti-SEB MAb, enzymes, and HRP, and combinations thereof.

8. The process of claim 1, wherein the biomolecules are selected from the group consisting of carbohydrates, sugars, dextrane, PEG, and poly-L-lysine and combinations thereof.

9. The process of claim 1, wherein the device is a CMOS device.

10. A microarray for a multiplex assay, comprising:

(a) a microarray having on a surface an array of addressable microelectrodes having an exposed surface;

(b) an electropolymerized polymer attached to the exposed surface of the microelectrodes; and

(c) a plurality of different non-functionalized biomolecules serially-attached to the polymer at different microelectrodes of the microarray, wherein the biomolecules are selected from the group consisting of antibodies, antigens, proteins, Ricin, anti-SEB MAb, enzymes, and HRP and combinations thereof.

11. The microarray of claim 10, wherein the microarray is a CMOS device.

12. The microarray of claim 10, wherein the polymer is made by electropolymerizing a monomer that is selected from the group consisting of pyrrole and functionalized pyrrole and combinations thereof.

13. The microarray of claim 10, wherein the polymer is made by electropolymerizing a monomer that is selected from the group consisting of thiophenol, aniline, phenylene sulfide, monomers electropolymerizing to a conducting polymer, phenylenediamine, diaminohaphthalene, phenol, monomers electropolymerizing to a nonconducting polymer and phenolic derivatives, and combinations thereof.

14. A method of making a multiplex microarray of serially attached different oligonucleotides on a microarray, comprising:

(a) blocking a microarray with a blocking protein, wherein the microarray is a device having an array of addressable microelectrodes on a surface of the device;

(b) electropolymerizing a monomer on one or more microelectrodes of the microarray to form a polymer coating on the one or more microelectrodes, wherein an electropolymerizing solution containing the monomer does not contain any of the oligonucleotides;

(c) exposing the microarray with an oligonucleotide solution containing an oligonucleotide for attachment to the polymer coating on the one or more electrodes;

(d) blocking the microarray with a blocking protein; and

(e) repeating steps (b) through (d) for each different oligonucleotide to be attached to the microarray, thereby making the microarray of different oligonucleotides.

15. The process of claim 14, wherein steps (a) and (d) further comprise:

washing the microarray to remove the blocking solution.

16. The process of claim 14, wherein step (b) further comprises:

washing the microarray to remove the electropolymerizing solution.

17. The process of claim 14, wherein step (c) further comprises:

washing the microarray to remove the biomolecular solution.

18. The process of claim 14, wherein the monomer is selected from the group consisting of pyrrole and functionalized pyrrole and combinations thereof.

19. The process of claim 14, wherein the monomer is selected from the group consisting of thiophenol, aniline, phenylene sulfide, monomers electropolymerizing to a conducting polymer, phenylenediamine, diaminohaphthalene, phenol, monomers electropolymerizing to a nonconducting polymer and phenolic derivatives, and combinations thereof.

20. A microarray for a multiplex oligonucleotide hybridization assay, comprising:

(a) a microarray having on a surface an array of addressable microelectrodes having an exposed surface;

(b) an electropolymerized polymer attached to the exposed surface of the microelectrodes; and

(c) a plurality of different preformed oligonucleotides serially-attached to the polymer at different microelectrodes of the microarray, wherein the different preformed oligonucleotides are not synthesized in situ on the microarray.