Compositions, methods, kits and devices for molecular analysis

Patent metadata

Field	Value
Publication number	US-11619608-B2
Application number	US-201816615804-A
Country	US
Kind code	B2
Filing date	May 22, 2018
Priority date	May 22, 2017
Publication date	Apr 4, 2023
Grant date	Apr 4, 2023

Abstract

Provided herein is an electrophoresis separation medium comprising: (a) a non-crosslinked or sparsely cross-linked polymer or copolymer; (b) one or more denaturant compounds, in an amount sufficient to inhibit re-naturation of single stranded polynucleotides; (c) an aqueous solvent; (d) optionally, a wall-coating material suited to inhibition of electroosmotic flow; and (e) optionally, an organic water miscible solvent such as DMSO or acetonitrile, wherein the electrophoresis separation medium exhibits functional stability for at least seven days at 23° C. Also provided herein are sieving compositions, including polymer-based sieving compositions, for molecular sieving as well as related kits, devices and methods of use. Such compositions can be useful for separation of biomolecules such as nucleic acids, proteins, glycoproteins and glycans.

First claim

What is claimed is:

1. An electrophoresis separation medium comprising:
(a) a sieving component comprising at least one polymer or copolymer;
(b) a denaturant comprising one or more of methylurea, ethylurea, a dimethylurea, or a diethylurea, and
(c) an aqueous solvent or aqueous buffer;
wherein the electrophoresis separation medium exhibits functional stability for capillary electrophoresis after storage at a temperature of at least about 23° C. for at least 14 days.

2. The medium of claim 1 , wherein the at least one polymer or copolymer is a non-crosslinked or sparsely cross-linked polymer or copolymer.

3. The medium of claim 1 , wherein at least one polymer or copolymer is crosslinked.

4. The medium of claim 1 , wherein the at least one polymer or copolymer is an uncharged water-soluble silica-adsorbing polymer or copolymer, a non-crosslinked acrylamide polymer or copolymer, a cellulose polymer or copolymer, a poly(alkylene oxide) polymer or copolymer, a polysaccharide, or a triblock copolymer.

5. The medium of claim 1 , further comprising one or more of a compound capable of adjusting refractive index of the medium, a compound capable of slowing down the re-naturation kinetics of single-stranded DNA, or a compound capable of enhancing resolution of molecules by the medium.

6. The medium of claim 1 , wherein the medium includes one or more agents selected from the group consisting of proline, histidine, betaine, trehalose, acetonitrile, imidazole, DMSO, N-methyl-2-pyrrolidinone, 3-(1-pyridinio)-1-propanesulfonate, 2-N,N,N-tri-n-butylammonium acetate, 1,3-dimethylurea, 1,3-diethylurea, 1,1-dimethylurea, and 1,1-diethylurea.

7. The medium of claim 1 , further comprising a saccharide or its derivatives, a sugar alcohol or polyol or its derivatives, a sugar or a sugar isomer or its derivatives, a pentose sugar, a hexose sugar, a starch, a carbohydrate, or a starch hydrolysate.

8. The medium of claim 1 , further comprising sucrose.

9. The medium of claim 8 , wherein the sucrose is included at 10 wt. % to 50 wt. % of the medium.

10. The medium of claim 1 , wherein the at least one polymer or copolymer comprises an N-substituted acrylamide polymer selected from the group consisting of N-substituted dimethylacrylamide, N-substituted diethylacrylamide, N-acryloyl-aminoethoxyethanol acrylamide, N-allyl glucose, and combinations thereof.

11. An electrophoresis separation medium comprising:
(a) a sieving component comprising at least one polymer or copolymer, wherein the at least one polymer or copolymer comprises an N-substituted acrylamide polymer selected from the group consisting of N-substituted dimethylacrylamide, N-substituted diethylacrylamide, N-acryloyl-aminoethoxyethanol acrylamide, N-allyl glucose, and combinations thereof;
(b) a denaturant comprising one or more of methylurea, ethylurea, a dimethylurea, or a diethylurea, and
(c) an aqueous solvent or aqueous buffer;
wherein the electrophoresis separation medium exhibits functional stability for capillary electrophoresis after storage at a temperature of at least about 23° C. for at least 14 days.

12. The medium of claim 11 , wherein the at least one polymer or copolymer is a non-crosslinked or sparsely cross-linked polymer or copolymer.

13. The medium of claim 11 , wherein at least one polymer or copolymer is crosslinked.

14. The medium of claim 11 , wherein the medium further comprises one or more agents that are selected from the group consisting of proline, histidine, betaine, trehalose, acetonitrile, imidazole, DMSO, N-methyl-2-pyrrolidinone, 3-(1-pyridinio)-1-propanesulfonate, 2-N,N,N-tri-n-butylammonium acetate, 1,3-dimethylurea, 1,3-diethylurea, ethylurea, methylurea, 1,1-dimethylurea, and 1,1-diethylurea.

15. The medium of claim 11 , further comprising sucrose included at 10 wt. % to 50 wt. % of the medium.

16. An electrophoresis separation medium comprising:
(a) a sieving component comprising at least one polymer or copolymer;
(b) sucrose included at 10 wt. % to 50 wt. % of the medium; and
(c) an aqueous solvent or aqueous buffer;
wherein the electrophoresis separation medium exhibits functional stability for capillary electrophoresis after storage at a temperature of at least about 23° C. for at least 14 days.

17. The medium of claim 16 , wherein the at least one polymer or copolymer is a non-crosslinked or sparsely cross-linked polymer or copolymer.

18. The medium of claim 16 , wherein at least one polymer or copolymer is crosslinked.

19. The medium of claim 16 , wherein the at least one polymer or copolymer is an uncharged water-soluble silica-adsorbing polymer or copolymer, a non-crosslinked acrylamide polymer or copolymer, a cellulose polymer or copolymer, a poly(alkylene oxide) polymer or copolymer, a polysaccharide, or a triblock copolymer.

20. The medium of claim 16 , further comprising one or more agents selected from the group consisting of proline, histidine, betaine, trehalose, acetonitrile, imidazole, DMSO, N-methyl-2-pyrrolidinone, 3-(1-pyridinio)-1-propanesulfonate, 2-N,N,N-tri-n-butylammonium acetate, 1,3-dimethylurea, 1,3-diethylurea, ethylurea, methylurea, 1,1-dimethylurea, and 1,1-diethylurea.

Assignees

• Life Technologies Corp
• Integenx Inc

Inventors

• Liu Jen-Kuei
• Solanki Bharti
• Wenz Hans Michael
• Mercer Frank
• Akin Cevat
• Karger Achim
• Barron Annelise
• Mack Scott

Classifications

• C08L2203/02

  for biomedical use · CPC title

• C08L2205/025

  containing two or more polymers of the same hierarchy C08L, and differing only in parameters such as density, comonomer content, molecular weight, structure · CPC title

• G01N27/44747 Primary

  Composition of gel or of carrier mixture · CPC title

• C08L33/26

  Homopolymers or copolymers of acrylamide or methacrylamide · CPC title