IP Library › Granted Patent US 12,624,384
Granted Patent B2
US 12,624,384 · App. 18/542,404 · Granted May 12, 2026

Compositions, methods, and systems for bead formation using improved polymers

Inventors: Joshua Delaney (Oakland, CA); Shalini Gohil (Castro Valley, CA); Christopher Hindson (Livermore, CA); Adam Lowe (Pleasanton, CA); Andrew D. Price (Hayward, CA); Joseph Francis Shuga (Pleasanton, CA)
Assignee: 10x Genomics, Inc.
C12Q1/6806B01J13/0065B01J13/18C08J3/075C08J2333/00C12Q1/6869
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Quick Facts
Patent No.
US 12,624,384
App. No.
18/542,404
Granted
May 12, 2026
Kind
B2
Abstract

The present disclosure provides systems and methods for making a hydrogel comprising a cell, cell nucleus, or one or more components derived from a cell or cell nucleus. A method for making a hydrogel may comprise providing a cell or cell nucleus, a first polymer, wherein the first polymer comprises a plurality of first crosslink precursors, each of the plurality of first crosslink precursors comprising an azide group; providing a second polymer, wherein the second polymer comprises a plurality of second crosslink precursors, each of the plurality of second crosslink precursors comprising an alkyne group; and crosslinking the first polymer and the second polymer via a reaction between a first section of the first crosslink precursors and a second section of the second crosslink precursors, thereby providing the hydrogel comprising the cell or cell nucleus.

Claims (25)

1 . A composition, comprising:

a partition comprising a matrix encapsulating a cell or cell nucleus, wherein the matrix is diffusively permeable to chemical reagents, wherein:

i) the matrix comprises two or more crosslinked polymers, wherein at least one polymer of the two or more crosslinked polymers is selected from the group consisting of a polyolefin, an olefin copolymer, an acrylic, a vinyl polymer, a polyamide, a polyimide, a formaldehyde resin, a polyurethane, a cellulosic, a thermoplastic elastomer, and a thermoplastic polyurethane; and

ii) the two or more crosslinked polymers comprise a triazole group, a dihydropyridazine group, a Diels-Alder adduct, or an isoxazoline group.

2 . The composition of claim 1 , wherein the partition is a droplet is among a plurality of droplets, wherein the droplet is fluidically isolated from other droplets in the plurality of droplets.

3 . The composition of claim 1 , wherein the partition is a well.

4 . The composition of claim 1 , wherein the two or more crosslinked polymers are attached to an oligonucleotide.

5 . The composition of claim 4 , wherein the oligonucleotide comprises a poly-T sequence or a poly-A sequence.

6 . The composition of claim 1 , wherein the composition is copper-free.

7 . The composition of claim 1 , wherein the matrix comprises a labile bond selected from a disulfide bond, a carbamate bond, and a peptide bond.

8 . The composition of claim 7 , wherein the labile bond is a peptide bond.

9 . The composition of claim 8 , wherein the labile bond is a carbamate linkage.

10 . The composition of claim 1 , wherein the partition further comprises a barcode molecule.

11 . The composition of claim 10 , wherein the barcode molecule is a nucleic acid barcode molecule.

12 . The composition of claim 11 , wherein the nucleic acid barcode molecule is attached to a bead in the partition.

13 . The composition of claim 1 , wherein the chemical reagents are selected from nucleic acids, enzymes, and potential cell binding ligands.

14 . A method of forming a matrix comprising:

(a) combining under copper-free click chemistry reaction conditions in a partition among a plurality of partitions: (i) a cell or cell nucleus, and (ii) two or more polymers, wherein at least one polymer of said two or more polymers is selected from the group consisting of a polyolefin, an olefin copolymer, an acrylic, a vinyl polymer, a polyamide, a polyimide, a formaldehyde resin, a polyurethane, a cellulosic, a thermoplastic elastomer, and a thermoplastic polyurethane; and

(b) forming crosslinks between the two or more polymers, thereby generating a matrix encapsulating the cell or cell nucleus.

15 . The method of claim 14 , wherein (b) comprises using (i) strain- promoted azide/dibenzocyclooctyne-amine (DBCO) click chemistry; (ii) inverse electron demand Diels-Alder (IED-DA) tetrazine/trans-cyclooctene (TCO) click chemistry; (iii) inverse electron demand Diels-Alder (IED-DA) tetrazine/norbornene click chemistry; (iv) Diels-Alder maleimide/furan click-chemistry; (v) Staudinger ligation; or (vi) nitrile-oxide/norbonene cycloaddition click chemistry to form crosslinks between the two or more polymers.

16 . The method of claim 14 , further comprising degrading the matrix.

17 . The method of claim 16 , wherein the matrix comprises a carbamate bond and the method further comprises degrading the matrix using a polyamine and heat to degrade the carbamate bond.

18 . The method of claim 16 , wherein the matrix comprises a peptide bond and the method further comprises degrading the matrix using a protease to degrade the peptide bond.

19 . The method of claim 14 , wherein the partition among the plurality of partitions is selected from a well and a droplet.

20 . The method of claim 14 , further comprising using a barcode molecule in the partition and a nucleic acid from the cell or cell nucleus to generate a barcoded nucleic acid molecule.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 5, 2024
From: DELANEY, JOSHUA; GOHIL, SHALINI; HINDSON, CHRISTOPHER; LOWE, ADAM; PRICE, ANDREW D.; SHUGA, JOSEPH FRANCIS
To: 10X GENOMICS, INC.
Reel/Frame 068186/0891 →
Continuity (8)
Continuation 17861503 · Jul 11, 2022
Continuation 17009352 · Sep 1, 2020
Continuation 16374112 · Apr 3, 2019
Continuation In Part 16178430 · Nov 1, 2018
Continuation PCTUS2018054458 · Oct 4, 2018
Provisional Application 62687161 · Jun 19, 2018
Provisional Application 62568021 · Oct 4, 2017
Related Publication 20240271183A1 · Aug 15, 2024
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