Neutralization and containment of redox species produced by circumferential electrodes

• US 10,525,436 B2
• Filed: 07/13/2016
• Issued: 01/07/2020
• Est. Priority Date: 04/15/2005
• Status: Active Grant

• Current Assignee: CustomArray, Inc. (Genscript Biotech Corp.)
• Original Assignee: CustomArray, Inc. (Genscript Biotech Corp.)
• Inventors: Maurer, Karl, Cooper, Jr., John J, Fujii, H Sho, Leonetti, Joseph
• Primary Examiner(s): Wilkins, III, Harry D
• Application Number:
  US15/208,606
• Publication Number:
  US 20160354751A1
• Time in Patent Office: 1,273 Days
• Field of Search: None
• 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 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• 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 2219/00675 In-situ synthesis on the substrate
• CPC Class Codes: B01J 2219/00713 Electrochemical synthesis
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00731 Saccharides
• CPC Class Codes: B01J 2219/00736 Non-biologic macromolecules, e.g. polymeric compounds
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C25B 11/02 characterised by shape or form
• CPC Class Codes: C25B 3/29 Coupling reactions
• CPC Class Codes: C25B 9/17 Cells comprising dimensionally-stable non-movable electrodes; Assemblies of constructional parts thereof

Abstract

There is disclosed an electrode array architecture employing continuous and discontinuous circumferential electrodes. There is further disclosed a process for the neutralization of acid generated at anode(s) by base generated at cathode(s) circumferentially located to each other so as to confine a region of pH change. The cathodes can be displayed as concentric rings (continuous) or as counter electrodes in a cross pattern (discontinuous). In this way reagents, such as acid, generated in a center electrode are countered (neutralized) by reagents, such as base, generated at the corners or at the outer ring.

Claims

1. A method for electrochemical oligomer synthesis on a dense electrode array comprising:

(a) providing a dense electrode array including a plurality of cells and a surface, where each cell of the plurality of cells includes an anode and a circumferential cathode, where each of the anodes are separately addressable electrodes, where a porous reaction layer is adsorbed to the surface, where the porous reaction layer is formed from one or more materials selected from the group consisting of a disaccharide, and a monosaccharide;

(b) contacting the porous reaction layer with a bathing solution; and

(c) bonding a first nucleotide having at least one protected chemical functional group to the porous reaction layer;

(d) conducting a deblocking step by addressing each active anode with a first current or a first voltage to generate an acid, where each active anode is surrounded by a cathode that is biased with a second current or a second voltage, where the cathode produces a base to neutralize the acid, where the cathode is separated from each active anode by an electrode margin, where an insulating material is present in the electrode margin, where the at least one protected chemical functional group is removed by the action of the acid forming at least one deprotected chemical functional group; and

(e) bonding a second nucleotide having at least one protected chemical functional group to the at least one deprotected chemical functional group.

2. The method of claim 1, where step (d) addresses each active anode with the first current.

3. The method of claim 1, where step (d) addresses the cathode with the second current.

4. The method of claim 1, where at least two neighboring electrodes are arranged in a pattern selected from the group consisting of nearest neighbor, neighbor electrodes, border of neighbor electrodes, border of nearest-neighbor, one closest available electrode, and electrode that is not approach.

5. The method of claim 1, where the insulating material is no more than 10 microns from the outer edge of each anode to the inner edge of the cathode.

6. A method for electrochemical oligomer synthesis on a dense electrode array comprising:

(a) providing a dense electrode array including a plurality of cells and a surface, where each cell of the plurality of cells includes an anode and a cathode, where the cathode forms an outer ring around each anode, where each of the anodes are separately addressable, where a porous reaction layer comprising a disaccharide and/or a monosaccharide is adsorbed to the surface;

(b) applying a bathing solution to the porous reaction layer;

(c) bonding a first nucleotide having at least one protected chemical functional group to the porous reaction layer;

(d) addressing one or more anodes with a first current or a first voltage to generate an acid;

(e) applying a second current or a second voltage to the cathode to generate a base, where the base neutralizes the acid, where the cathode is separated from each active anode by an electrode margin, where an insulating material is present in the electrode margin, where the at least one protected chemical functional group is removed by the action of the acid forming at least one deprotected chemical functional group; and

(f) bonding a second nucleotide having at least one protected chemical functional group to the at least one deprotected chemical functional group.

7. The method of claim 6, where the thickness of the insulating material is between:

a lower limit of 10 microns; and

an upper limit of 50 microns.

8. The method of claim 6, where an outer edge of the outer ring of a first cell is separated from an outer edge of the outer ring of a second cell by between:

a lower limit of 33 microns; and

an upper limit of 50 microns.

9. The method of claim 6, where the cell diameter is 44 microns.

10. The method of claim 6, where a center point of a first cell is separated from a center point of a second cell by 75 microns.

11. The method of claim 6, where step (d) addresses each active anode with the first current.

12. The method of claim 6, where step (e) applies the second current to the cathode.

13. A method for electrochemical oligomer synthesis on a dense electrode array comprising:

(a) providing a dense electrode array including a plurality of cells and a surface, where each cell of the plurality of cells includes a cathode and an anode, where the anode forms an outer ring around each cathode, where each of the cathodes are separately addressable, where a porous reaction layer comprising a disaccharide and/or a monosaccharide is adsorbed to the surface;

(b) applying a bathing solution to the porous reaction layer;

(c) bonding a first nucleotide having at least one protected chemical functional group to the porous reaction layer;

(d) addressing one or more cathodes with a first current or a first voltage to generate a base;

(e) applying a second current or a second voltage to the anode to generate an acid, where the acid neutralizes the base, where the anode is separated from each active cathode by an electrode margin, where an insulating material is present in the electrode margin, where the at least one protected chemical functional group is removed by the action of the acid forming at least one deprotected chemical functional group; and

(f) bonding a second nucleotide having at least one protected chemical functional group to the at least one deprotected chemical functional group.

14. The method of claim 13, where the thickness of the insulating material is between:

a lower limit of 10 microns; and

an upper limit of 50 microns.

15. The method of claim 13, where an outer edge of the outer ring of a first cell is separated from an outer edge of the outer ring of a second cell by between:

a lower limit of 33 microns; and

an upper limit of 50 microns.

16. The method of claim 13, where the cell diameter is 44 microns.

17. The method of claim 13, where a center point of a first cell is separated from a center point of a second cell by 75 microns.