IP Library Granted Patent US 8,734,630
Granted Patent B2
US 8,734,630 · App. 13/615,417 · Granted May 27, 2014

High speed, high resolution compositions, methods, and kits for capillary electrophoresis

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Quick Facts
Patent No.
US 8,734,630
App. No.
13/615,417
Granted
May 27, 2014
Kind
B2
Abstract

The invention provides compositions, methods and kits for high speed, high resolution of analytes by capillary electrophoresis starting with uncoated capillaries. The compositions comprise a sieving component, comprising a non-crosslinked acrylamide polymer, and a surface interaction component, comprising at least one uncharged and non-crosslinked water-soluble silica-adsorbing polymer. Methods for employing the novel compositions in capillary electrophoresis are provided. Kits comprising the novel compositions for use in the novel methods are also provided.

Claims (54)

1. A method for separating analytes by capillary electrophoresis comprising:

providing a capillary electrophoresis apparatus having at least one capillary;

inserting into the at least one capillary a composition comprising:

(a) a sieving component comprising an acrylamide polymer having a weight average molecular weight greater than about 3,000,000 Da and less than or equal to about 6,400,000 Da; and

(b) a surface interaction component comprising one or more polymers selected from the group consisting of poly(meth)acrylamide, N,N-disubstituted polyacrylamide and N-substituted polyacrylamide, wherein said N-substituents are selected from the group consisting of C 1 to C 3 alkyl, halo-substituted C 1 to C 3 alkyl, methoxy-substituted C 1 to C 3 alkyl, and hydroxyl-substituted C 1 to C 3 alkyl;

wherein:

the sieving component and the surface interaction component differ in polymer chemical composition; and

the composition does not include a crosslinked polymer gel;

loading a sample of different sized analytes into the at least one capillary; and

separating the analytes by capillary electrophoresis.

2. The method of claim 1 , which is carried out in parallel with a plurality of uncoated capillaries.

3. The method of claim 1 , wherein the composition further comprises at least one denaturant.

4. The method of claim 3 , wherein the at least one denaturant is selected from the group consisting of at least one of formamide, urea and 2-pyrollidinone.

5. The method of claim 4 , wherein the at least one denaturant comprises urea.

6. A method for separating analytes by capillary electrophoresis comprising:

providing a capillary electrophoresis apparatus having a capillary electrophoresis element comprising:

(a) an uncoated capillary; and

(b) a composition for separating analytes located within the uncoated capillary, the composition comprising:

(i) a sieving component comprising a non-crosslinked acrylamide polymer having a weight average molecular weight greater than about 3,000,000 Da and less than or equal to about 6,400,000 Da; and

(ii) a surface interaction component; wherein the surface interaction component comprises a solution of non-crosslinked N,N-dimethylacrylamide;

wherein the sieving component and the surface interaction component differ in polymer chemical composition, and

wherein the capillary electrophoresis element does not include a crosslinked polymer gel;

injecting the analytes electrokinetically into the capillary electrophoresis element; and

separating the analytes by capillary electrophoresis.

7. The method of claim 6 , which is carried out in parallel with a plurality of uncoated capillaries.

8. The method of claim 7 , wherein the uncoated capillaries comprise silica, fused silica, quartz, silicate-based glass, phosphate glass, or alumina-containing glass.

9. The method of claim 7 , wherein the uncoated capillaries are plastic channel capillaries.

10. A method for separating analytes by capillary electrophoresis comprising:

providing a capillary electrophoresis apparatus having a capillary electrophoresis element comprising:

(a) an uncoated capillary;

(b) a composition for separating analytes located within the uncoated capillary, the composition comprising:

(i) a sieving component comprising a non-crosslinked acrylamide polymer having a weight average molecular weight greater than about 3,000,000 Da and less than or equal to about 6,400,000 Da; and

(ii) a surface interaction component comprising a solution of non-crosslinked N,N-dimethylacrylamide; and a denaturant comprising urea; and

wherein the capillary electrophoresis element does not include a crosslinked polymer gel;

injecting the analytes into the capillary electrophoresis element; and

separating the analytes by capillary electrophoresis.

11. The method of claim 10 , which is carried out in parallel with a plurality of capillaries.

12. The method of claim 11 , wherein the capillaries comprise silica, fused silica, quartz, silicate-based glass, phosphate glass, or alumina-containing glass.

13. The method of claim 11 , wherein the capillaries are plastic channel capillaries.

14. A method for separating analytes by capillary electrophoresis comprising:

inserting into an uncoated capillary having a first end and a second end a composition comprising:

(a) a sieving component comprising an uncrosslinked acrylamide polymer having a weight average molecular weight greater than about 3,000,000 Da and less than or equal to about 6,400,000 Da; and

(b) a surface interaction component comprising one or more non-crosslinked polymers selected from the group consisting of poly(meth)acrylamide, N,N-disubstituted polyacrylamide and N-substituted polyacrylamide, wherein said N-substituents are selected from the group consisting of C 1 to C 3 alkyl, halo-substituted C 1 to C 3 alkyl, methoxy-substituted C 1 to C 3 alkyl, and hydroxyl-substituted C 1 to C 3 alkyl;

wherein

(i) the sieving component and the surface interaction component differ in polymer chemical composition; and

(ii) the composition does not include a crosslinked polymer gel;

loading a sample of different sized analytes in the capillary; and

applying an electrical field between the first and second ends of the capillary configured to cause the different sized analytes in the sample to migrate through the capillary, thereby separating the analytes.

15. The method of claim 14 , wherein the composition further comprises at least one denaturant.

16. The method of claim 15 , wherein the at least one denaturant comprises urea.

17. The method of claim 14 , wherein the surface interaction non-crosslinked polymer is poly(N,N-dimethylacrylamide).

18. The method of claim 14 , which is carried out in parallel with a plurality of uncoated capillaries.

19. The method of claim 18 , wherein the uncoated capillaries comprise silica, fused silica quartz, silicate-based glass, phosphate glass, or alumina-containing glass.

20. The method of claim 18 , wherein the uncoated capillaries are plastic channel capillaries.

Assignments (4)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 28, 2014
From: VOSS, KARL O; LAU, ALDRICH N.K.
To: PE CORPORATION (NY)
Reel/Frame 032976/0721 →
MERGER Recorded May 28, 2014
From: APPLERA CORPORATION
To: APPLIED BIOSYSTEMS INC.
Reel/Frame 032977/0150 →
MERGER Recorded May 28, 2014
From: ATOM ACQUISITION CORPORATION; APPLIED BIOSYSTEMS INC.
To: APPLIED BIOSYSTEMS, LLC
Reel/Frame 032977/0240 →
CHANGE OF NAME Recorded May 28, 2014
From: PE CORPORATION (NY)
To: APPLERA CORPORATION
Reel/Frame 033036/0820 →