Devices and methods for synthesis

Patent metadata

Field	Value
Publication number	US-12534494-B2
Application number	US-202117458200-A
Country	US
Kind code	B2
Filing date	Aug 26, 2021
Priority date	Aug 28, 2020
Publication date	Jan 27, 2026
Grant date	Jan 27, 2026

Abstract

Provided herein are compositions, devices, systems and methods for electrochemical synthesis. Further provided are devices comprising addressable electrodes controlling polynucleotide synthesis (deprotection, extension, or cleavage, etc.) The compositions, devices, systems and methods described herein provide improved synthesis, storage, density, and retrieval of biomolecule-based information.

First claim

What is claimed is:

1 . A method for synthesizing a polynucleotide comprising:
providing a solid support, the solid support including:
a base layer comprising silicon,
an intermediate layer comprising an oxide, and
a top layer comprising platinum;
wherein the intermediate layer is positioned between the base layer and the top layer, and wherein the top layer comprises a plurality of voids configured to allow fluid communication of a solvent with the intermediate layer; contacting at least one nucleoside attached to the intermediate layer of the solid support with a protected nucleoside, wherein the protected nucleoside is configured to form a covalent bond with the at least one nucleoside to generate a protected polynucleotide; applying a voltage to a solvent in fluid communication with the protected polynucleotide, wherein the voltage results in deprotection of a terminal nucleoside of the protected polynucleotide, and wherein the voltage is delivered as at least 2 pulses; and repeating the contacting step and the applying step to synthesize the polynucleotide.

2 . The method of claim 1 , wherein the voltage is 0.5-2.5 volts.

3 . The method of claim 2 , wherein the total period of time for all pulses is less than 5 seconds.

4 . The method of claim 1 , wherein the pulse is no more than 50 milliseconds.

5 . The method of claim 1 , wherein the voltage is delivered as at least 100 pulses.

6 . The method of claim 1 , wherein the solvent comprises a composition for electrochemical acid generation.

7 . The method of claim 6 , wherein the solvent comprises hydroquinone, benzoquinone, or a mixture thereof.

8 . The method of claim 1 , wherein the protected polynucleotide comprises an acid-cleavable protecting group.

9 . The method of claim 1 , wherein the time between any two pulses is 10-2000 milliseconds.

10 . The method of claim 1 , wherein the polynucleotide is washed between pulses.

11 . A method for synthesizing a polynucleotide comprising:
(a) providing a solid support, the solid support comprising:
a base layer comprising silicon;
an intermediate layer comprising an oxide; and
a top layer, wherein the intermediate layer is positioned between the base layer and the top layer, and the top layer comprises a plurality of voids configured to allow fluid communication of a solvent with the intermediate layer; and
wherein the intermediate layer comprises a first plurality of protected polynucleotides;
(b) energizing at least one electrode in contact with the intermediate layer to electrochemically generate a deprotection reagent, wherein the deprotection reagent deprotects at least one polynucleotide of the first plurality of protected polynucleotides; (c) coupling at least one protected nucleoside to the at least one deprotected polynucleotide; and (d) repeating steps (a)-(c) to synthesize the polynucleotide.

12 . The method of claim 11 , wherein the one or more electrodes comprise at least one anode.

13 . The method of claim 11 , wherein the surface comprises a second region comprising a second plurality of protected polynucleotides.

14 . The method of claim 13 , wherein a pitch distance between the first region and the second region is no more than 1 micron.

15 . The method of claim 13 , wherein no more than 1% of the second plurality of protected polynucleotides is deprotected.

16 . The method of claim 13 , wherein the first plurality of protected polynucleotides and the second plurality of protected polynucleotides comprise protecting groups capable of removal with acid.

17 . The method of claim 11 , further comprising applying a voltage to a solvent in fluid communication with the first plurality of protected polynucleotides, wherein the voltage generates acid.

18 . The method of claim 13 , wherein the first region and the second region each comprise an addressable anode.

19 . The method of claim 11 , wherein a positive voltage is applied to the one or more electrodes, and no voltage or a negative voltage is applied to the cathode.

20 . The method of claim 18 , wherein the absolute voltage difference between the anode and cathode is no more than 2 volts.

Assignees

• Twist Bioscience Corp

Inventors

• Fernandez Andres
• Marsh Eugene P
• Habibian Maryam

Classifications

• B01J2219/00722

  Nucleotides · CPC title

• B01J2219/00713

  Electrochemical synthesis · CPC title

• B01J19/087

  employing electric or magnetic energy · CPC title

• B01J19/0046

  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 title

• C07H21/00 Primary

  Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids · CPC title