Serially deposited biomolecules

• US 10,591,476 B1
• Filed: 03/26/2018
• Issued: 03/17/2020
• Est. Priority Date: 01/20/2010
• Status: Active Grant

• Current Assignee: CustomArray, Inc. (Genscript Biotech Corp.)
• Original Assignee: CustomArray, Inc. (Genscript Biotech Corp.)
• Inventors: Cooper, John
• Primary Examiner(s): Crow, Robert T.
• Application Number:
  US15/936,298
• Time in Patent Office: 722 Days
• Field of Search: None
• 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 process of preparing two or more oligonucleotides on a microarray, comprising:

(a) electropolymerizing a monomer on one or more microelectrodes of a microarray using an electropolymerizing solution with one or more electropolymerizing parameters to form a first thickness on at least a first microelectrode and electropolymerizing the monomer with one or more different electropolymerizing parameters to form at least a second thickness on at least a second microelectrode, where the first microelectrode and the second microelectrode are selected from the one or more microelectrodes, where the first thickness is different than the second thickness, where the one or more electropolymerizing parameters are selected from the group consisting of potential, current and time of electropolymerizing;

(b) exposing the microarray to a first solution containing a first oligonucleotide for attachment to the first thickness on the one or more electrodes; and

(c) exposing the microarray to a second solution containing a second oligonucleotide for attachment to the second thickness on the one or more electrodes, where one or both the first thickness and the second thickness facilitates detection of one or both the first oligonucleotide and the second oligonucleotide.

2. The process of claim 1, further comprising applying a blocking protein prior to one or more of steps (a), (b) and (c).

3. The process of claim 2, further comprising washing the microarray to remove the blocking protein.

4. The process of claim 1, where step (a) further comprises washing the microarray to remove the electropolymerizing solution.

5. The process of claim 1, where step (b) further comprises washing the microarray to remove the first solution.

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

7. The process of claim 1, where the monomer is selected from the group consisting of thiophenol, aniline, phenylene sulfide, phenylenediamine, diaminohaphthalene, phenol, phenolic derivatives, and combinations thereof.

8. The process of claim 1, where the microarray is a CMOS device.

9. The process of claim 1, where the monomer is electropolymerized to a conducting polymer.

10. The process of claim 1, where the monomer is electropolymerized to a non conducting polymer.

11. A process of preparing a multiplex microarray of serially attached different oligonucleotides on a microarray, comprising:

(a) electropolymerizing a monomer on one or more microelectrodes of the microarray using an electropolymerizing solution with one or more electropolymerizing parameters to form a first thickness on a first microelectrode and electropolymerizing the monomer with one or more different electropolymerizing parameters to form at least a second thickness on a second microelectrode, where the first microelectrode and the second microelectrode are selected from the one or more microelectrodes, where the first thickness is different than the second thickness, where the one or more electropolymerizing parameters are selected from the group consisting of potential, current and time of electropolymerizing, where the electropolymerizing solution containing the monomer does not contain any oligonucleotides;

(b) exposing the microarray with a first oligonucleotide solution containing a first oligonucleotide for attachment to the first thickness on the first microelectrode; and

(c) exposing the microarray to a second oligonucleotide solution containing a second oligonucleotide for attachment to the second thickness on the second microelectrode, where one or both the first thickness and the second thickness facilitates detection of one or both the first oligonucleotide and the second oligonucleotide.

12. The process of claim 11, further comprising applying a blocking protein prior to one or more of steps (a), (b) and (c).

13. The process of claim 12, further comprising washing the microarray to remove the blocking protein.

14. The process of claim 11, where step (a) further comprises washing the microarray to remove the electropolymerizing solution.

15. The process of claim 11, where step (b) further comprises washing the microarray to remove the first oligonucleotide solution.

16. The process of claim 11, where the monomer is selected from the group consisting of pyrrole, functionalized pyrrole and combinations thereof.

17. The process of claim 11, where the monomer is selected from the group consisting of thiophenol, aniline, phenylene sulfide, phenylenediamine, diaminohaphthalene, phenol, phenolic derivatives, and combinations thereof.

18. The process of claim 11, where the microarray is a CMOS device.

19. The process of claim 11, where the monomer is electropolymerized to a conducting polymer.

20. The process of claim 11, where the monomer is electropolymerized to a non conducting polymer.