# US 10,246,533 B2 · Hydrophilic polymeric particles and methods for making and using same

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,246,533 B2
- Document ID: US10246533B2
- Status: Granted
- Kind: B2
- Application number: 14/970,192
- Filed: 2015-12-15
- Published: 2016-04-07
- Granted: 2019-04-02
- Earliest claimed: 2012-02-09
- Assignees: Life Technologies Corporation
- Applicants: Life Technologies Corporation; Life Technologies AS
- Inventors: Geir Fonnum; Grete Irene Modahl; Nini Hofslokken Kjus; Astrid Evenroed Molteberg; Diem Tran; Jo Aaserud; M. Talha Gokmen; Steven M. Menchen; Carl Fuller; Luisa Andruzzi; Wolfgang Hinz
- Web page: https://patentgrep.com/patent/US10246533B2

## CPC

- C08F 130/08: containing silicon
- C07F 7/1804: {Compounds having Si-O-C linkages (Si-O-acyl linkages C07F7/1896)}
- C07F 7/188: {by reactions involving the formation of Si-O linkages}
- C08F 2/20: with the aid of macromolecular dispersing agents
- C08F 2/26: anionic
- C08F 6/24: Treatment of polymer suspensions {(C08F6/001, C08F6/006, C08F6/008, C08F6/02, C08F6/04 take precedence)}
- C08F 8/12: Hydrolysis
- C08F 8/32: by reaction with amines
- C08F 8/34: Introducing sulfur atoms or sulfur-containing groups
- C08F 222/385: {Monomers containing two or more (meth)acrylamide groups, e.g. N,N'-methylenebisacrylamide}
- C08F 230/085: {the monomer being a polymerisable silane, e.g. (meth)acryloyloxy trialkoxy silanes or vinyl trialkoxysilanes}
- C08F 257/02: on to polymers of styrene or alkyl-substituted styrenes
- C08J 9/20: by suspension polymerisation in the presence of the blowing agent
- C09D 5/00: Coating compositions, e.g. paints, varnishes or lacquers, characterised by their physical nature or the effects produced {(electrically insulating plastics, resins or waxes H01B3/30)}; Filling pastes
- C12P 19/34: Polynucleotides, e.g. nucleic acids, oligoribonucleotides
- C12Q 1/686: Polymerase chain reaction [PCR]
- C12Q 1/6874: involving nucleic acid arrays, e.g. sequencing by hybridisation
- G01N 27/4145: {specially adapted for biomolecules, e.g. gate electrode with immobilised receptors}
- C08F 220/603: {and containing oxygen in addition to the carbonamido oxygen and nitrogen}
- C08F 2800/20: as weight or mass percentages
- C08J 2343/04: Homopolymers or copolymers of monomers containing silicon
- Y10T 428/2982: Particulate matter [e.g., sphere, flake, etc.]
- C08F: MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C07F: ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM (metal-containing porphyrins C07D487/22)
- C08J: WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H (working, e.g. shaping, of plastics B29)
- C09D: COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR (cosmetics A61K; processes for applying liquids or other fluent materials to surfaces, in general, B05D; staining wood B27K5/02; glazes or vitreous enamels C03C; natural resins, French polish, drying-oils, driers, turpentine, per se, C09F; polishing compositions other than French polish, ski waxes C09G; adhesives or use of materials as adhesives C09J; materials for sealing or packing joints or covers C09K3/10; materials for stopping leaks C09K3/12; processes for the electrolytic or electrophoretic production of coatings C25D)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

A method of forming a particle includes, in a disperse phase within an aqueous suspension, polymerizing a plurality of monomer units of a hydrophilic monomer having a hydrophobic protection group, thereby forming a polymeric particle including a plurality of the hydrophobic protection groups. The method further includes converting the polymeric particle to a hydrophilic particle.

## Claims

### Claim 1

A method of sequencing a polynucleotide, the method comprising:
providing a device including an array of wells, at least one well operatively connected to an ion sensitive field effect transistor and including a hydrogel particle, the hydrogel particle being attached to a polynucleotide,
applying a solution including nucleotides of a predetermined type to the device; and
observing an ionic response to the applying the solution,
the hydrogel particle formed by:
in a disperse phase within an aqueous suspension, polymerizing a plurality of monomer units of a hydrophilic monomer having a hydrophobic protection group, thereby forming a polymeric particle including a plurality of the hydrophobic protection groups; and
converting the polymeric particle to the hydrogel particle, wherein converting the polymeric particle includes removing at least a portion of the plurality of the hydrophobic protection groups from the polymeric particle.

### Claim 2 (depends on 1)

The method of claim 1, wherein removing at least a portion of the plurality of the hydrophobic protection groups includes acid cleaving at least a portion of the plurality of the hydrophobic protection groups from the polymeric particle.

### Claim 3 (depends on 1)

The method of claim 1, further comprising promoting a seed particle in the aqueous suspension to form the dispersed phase.

### Claim 4 (depends on 3)

The method of claim 3, wherein the seed particle includes a seed polymer, the method further comprising extracting the seed polymer after converting the polymeric particle.

### Claim 5 (depends on 4)

The method of claim 4, wherein the seed polymer is hydrophobic.

### Claim 6 (depends on 3)

The method of claim 3, wherein promoting the seed particle includes mixing a solvent and a promoting agent with the seed particle.

### Claim 7 (depends on 1)

The method of claim 1, wherein the hydrophilic monomer includes an acrylamide.

### Claim 8 (depends on 1)

The method of claim 1, wherein the hydrophobic protection group includes an organometallic moiety.

### Claim 9 (depends on 8)

The method of claim 8, wherein the organometallic moiety forms a silyl ether functional group.

### Claim 10 (depends on 1)

The method of claim 1, wherein polymerizing the plurality of monomer units further includes mixing a crosslinker with the hydrophilic monomer having a hydrophobic protection group.

### Claim 11 (depends on 10)

The method of claim 10, wherein the crosslinker is a divinyl crosslinker.

### Claim 12 (depends on 1)

The method of claim 1, further comprising activating the hydrogel particle.

### Claim 13 (depends on 12)

The method of claim 12, further comprising binding an oligonucleotide to the activated hydrogel polymer.

### Claim 14 (depends on 13)

The method of claim 13, wherein binding includes nucleophilic substitution and the oligonucleotide is a nucleophile-terminated oligonucleotide.

### Claim 15 (depends on 13)

The method of claim 13, further comprising hybridizing a polynucleotide to the oligonucleotide.

### Claim 16 (depends on 15)

The method of claim 15, further comprising amplifying the polynucleotide into a plurality of complementary polynucleotides by extending the oligonucleotide, thereby generating a hydrogel particle including a plurality of attached polynucleotides.

### Claim 17

A method for nucleotide incorporation, comprising:
providing a hydrogel particle, the particle being attached to a nucleic acid duplex including a template nucleic acid hybridized to a primer, the duplex being bound to a polymerase;
contacting the hydrogel particle with one or more nucleotides; and
incorporating at least one nucleotide onto the end of the primer using the polymerase,
the hydrogel particle formed by:
in a disperse phase within an aqueous suspension, polymerizing a plurality of monomer units of a hydrophilic monomer having a hydrophobic protection group, thereby forming a polymeric particle including a plurality of the hydrophobic protection groups; and
converting the polymeric particle to the hydrogel particle, wherein converting the polymeric particle includes removing at least a portion of the plurality of the hydrophobic protection groups from the polymeric particle.

### Claim 18 (depends on 17)

The method of claim 17, wherein removing at least a portion of the plurality of the hydrophobic protection groups includes acid cleaving at least a portion of the plurality of the hydrophobic protection groups from the polymeric particle.