Hydrophilic polymeric particles and methods for making and using same

Patent metadata

Field	Value
Publication number	US-10246533-B2
Application number	US-201514970192-A
Country	US
Kind code	B2
Filing date	Dec 15, 2015
Priority date	Feb 9, 2012
Publication date	Apr 2, 2019
Grant date	Apr 2, 2019

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.

First claim

What is claimed is:

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.

2. 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.

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

4. 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.

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

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

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

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

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

10. 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.

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

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

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

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

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

16. 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.

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.

18. 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.

Assignees

• Life Technologies Corp
• Life Tech As

Inventors

• Fonnum Geir
• Modahl Grete Irene
• Kjus Nini Hofslokken
• Molteberg Astrid Evenroed
• Tran Diem
• Aaserud Jo
• Gokmen M Talha
• Menchen Steven M
• Fuller Carl
• Andruzzi Luisa
• Hinz Wolfgang

Classifications

• C08F130/08 Primary

  containing silicon · CPC title

• C12Q1/686

  Polymerase chain reaction [PCR] · CPC title

• C08F6/24

  Treatment of polymer suspensions {(C08F6/001, C08F6/006, C08F6/008, C08F6/02, C08F6/04 take precedence)} · CPC title

• C08J9/20

  by suspension polymerisation in the presence of the blowing agent · CPC title

• C08F222/385

  Monomers containing two or more (meth)acrylamide groups, e.g. N,N'-methylenebisacrylamide · CPC title