Emulsion compositions and methods of their use

Patent metadata

Field	Value
Publication number	US-11845980-B2
Application number	US-202217665170-A
Country	US
Kind code	B2
Filing date	Feb 4, 2022
Priority date	Jan 30, 2017
Publication date	Dec 19, 2023
Grant date	Dec 19, 2023

Abstract

Emulsion compositions are provided herein. Also provided herein are kits containing one or more emulsion compositions or components for making such emulsion compositions. Also provided herein are methods of using such emulsion compositions, such as for amplification of target nucleic acids in emulsion droplets.

First claim

What is claimed is:

1. An emulsion comprising a water-in-oil emulsion droplets, the droplets comprising:
water;
a plurality of cyclodextrin molecules, wherein at least some of the cyclodextrin molecules comprise 2-hydroxypropyl α-cyclodextrin, 2-hydroxypropyl β-cyclodextrin, or 2-hydroxypropyl γ-cyclodextrin; and
a plurality of amphiphilic polymer molecules, wherein individual amphiphilic polymer molecules of the plurality comprise a hydrophobic moiety and a hydrophilic moiety, wherein the amphiphilic polymer molecules are poloxamers;
wherein at least a portion of the cyclodextrin molecules and the amphiphilic polymer molecules form a thermally-reversible hydrogel that is in sol form at at least one temperature between 0-70° C., wherein the thermally-reversible hydrogel comprises a plurality of polypseudorotaxane complexes, and
wherein the droplets are present in a continuous oil phase.

2. The emulsion of claim 1 , wherein a mass ratio of cyclodextrin molecules to amphiphilic polymer molecules in the droplets is from about 1:4 to about 4:1.

3. The emulsion of claim 1 , wherein a mass ratio of cyclodextrin molecules to amphiphilic polymer molecules in the droplets is 1:1.

4. The emulsion of claim 1 , wherein the concentration of cyclodextrin molecules in the droplets is from 0.5% to 4% mass per volume (m/v).

5. The emulsion of claim 4 , wherein the concentration of cyclodextrin molecules is 2% mass per volume (m/v).

6. The emulsion of claim 4 , wherein the poloxamers are poloxamer 188.

7. The emulsion of claim 1 , wherein the cyclodextrin molecules comprise 2-hydroxypropyl α-cyclodextrin.

8. The emulsion of claim 1 , wherein the cyclodextrin molecules comprise 2-hydroxypropyl β-cyclodextrin.

9. The emulsion of claim 1 , wherein the cyclodextrin molecules comprise 2-hydroxypropyl γ-cyclodextrin.

10. The emulsion of claim 1 , wherein the poloxamers are poloxamer 188.

11. The emulsion of claim 1 , wherein the droplets further comprise a thermostable DNA-dependent DNA polymerase or an RNA-dependent DNA polymerase.

12. The emulsion of claim 1 , wherein the droplets further comprise nucleic acid.

13. The emulsion of claim 1 , wherein the droplets comprise a thermally reversible hydrogel.

14. The emulsion of claim 1 , wherein the droplets are present in a continuous fluorocarbon oil phase.

15. The emulsion of claim 14 , wherein the continuous fluorocarbon oil phase comprises a fluorous oil and a surfactant.

16. The emulsion of claim 15 , wherein the surfactant is a fluorosurfactant.

17. The emulsion of claim 16 , wherein the fluorosurfactant is a block co-polymer comprising a perfluorinated polyether block and a polyethylene glycol block.

18. A method of forming an emulsion according to claim 1 , the method comprising:
combining a continuous oil phase, an aqueous solution, a plurality of cyclodextrin molecules, and a plurality of amphiphilic polymer molecules, under conditions wherein at least a portion of the cyclodextrin molecules and the amphiphilic polymer molecules form a plurality of polypseudorotaxane complexes, thereby forming an emulsion comprising water-in-oil emulsion droplets.

19. A method of using an emulsion comprising water-in-oil emulsion droplets, the method comprising:
providing the emulsion according to claim 1 ;
heating the droplets to a temperature of between about 50° C. to about 100° C.;
cooling the droplets to a temperature below about 50° C.; and
optionally repeating the heating and cooling between 1-50 times.

Assignees

• Bio Rad Laboratories Inc

Inventors

• Xue Yi
• Ritchey Joshua
• Meltzer Robert

Classifications

• C12Q1/6848 Primary

  characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction · CPC title

• C12Q1/686

  Polymerase chain reaction [PCR] · CPC title

• C12Q1/6846 Primary

  Common amplification features · CPC title

• C12Q1/6816

  characterised by the detection means (C12Q1/6804 takes precedence) · CPC title

• C12Q2527/125

  Specific component of sample, medium or buffer · CPC title