High speed, high resolution compositions, methods and kits for capillary electrophoresis
View Patent ↗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.
1. A high speed, high resolution composition for separating analytes by capillary electrophoresis comprising:
a sieving component comprising a non-crosslinked hydrophilic acrylamide polymer having a molecular weight greater than about 1,000,000 Daltons (Da) and less than or equal to about 3,000,000 Da; and
a surface interaction component comprising one or more non-crosslinked hydrophobic acrylamide polymers;
wherein the sieving component and the surface interaction component differ in polymer chemical composition;
the composition has a viscosity of less than 1,000 centipoise at 25° C.; and
the composition does not include a crosslinked polymer gel.
2. The composition of claim 1 , wherein the composition has a viscosity of less than 600 centipoise at 25° C.
3. The composition of claim 1 , wherein the one or more non-crosslinked hydrophobic acrylamide polymers comprises poly(N,N-dimethylacrylamide).
4. The composition of claim 1 , further comprising at least one denaturant.
5. The composition of claim 4 , wherein the at least one denaturant is selected from the group consisting of at least one of formamide, urea, and 2-pyrollidinone.
6. The composition of claim 5 , wherein the at least one denaturant comprises urea.
7. The composition of claim 1 , wherein the one or more hydrophobic acrylamide polymers are selected from the group consisting of 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, and —CH 2 CONH 2 , or the N-substituents of the N,N disubstituted polyacrylamide are taken together with the nitrogen to form a cyclic moiety having a structure of one of the following formulae:
8. The composition of claim 1 wherein the non-crosslinked hydrophilic acrylamide polymer is polyacrylamide.
9. The composition of claim 1 , wherein the one or more hydrophobic acrylamide polymers are selected from the group consisting of 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.
10. A composition for separating analytes by capillary electrophoresis comprising:
a sieving component comprising a non-crosslinked polyacrylamide polymer having a molecular weight greater than about 1,000,000 Daltons (Da) and less than or equal to about 3,000,000 Da; and having a viscosity of less than 10,000 centipoise at 25° C.;
a surface interaction component comprising poly(N,N-dimethylacrylamide);
wherein the sieving component and the surface interaction component differ in polymer chemical composition; and
wherein the composition does not include a crosslinked polymer gel.
11. The composition of claim 10 , wherein the viscosity is less than 5,000 centipoise at 25° C.
12. The composition of claim 10 , wherein the viscosity is less than 1,000 centipoise at 25° C.
13. The composition of claim 10 , wherein the viscosity is less than 600 centipoise at 25° C.
14. The composition of claim 10 further comprising:
a denaturant comprising urea.
15. The composition of claim 10 , wherein the non-crosslinked polyacrylamide polymer is linear polyacrylamide.
16. A capillary electrophoresis element comprising:
an uncoated capillary;
a composition for separating analytes located within the uncoated capillary, the composition comprising: a sieving component comprising a uncrosslinked hydrophilic acrylamide polymer having a molecular weight greater than about 1,000,000 Daltons (Da) and less than or equal to about 3,000,000-Da;
a surface interaction component comprising one or more non-crosslinked hydrophobic acrylamide polymers;
wherein the surface interaction component comprises a solution of the one or more non-crosslinked hydrophobic polymers;
the sieving component and the surface interaction component differ in polymer chemical composition;
the composition has a viscosity of less than 1,000 centipoise at 25° C.; and
wherein the capillary electrophoresis element does not include a crosslinked polymeric gel.
17. The capillary electrophoresis element of claim 16 , wherein the composition has a viscosity of less than 5000 centipoise at 25° C.
18. The capillary electrophoresis element of claim 16 , wherein the composition has a viscosity of less than 1000 centipoise at 25° C.
19. The capillary electrophoresis element of claim 16 , wherein the composition has a viscosity of less than 600 centipoise at 25° C.
20. The capillary electrophoresis element of claim 16 , wherein the one or more non-crosslinked hydrophobic acrylamide polymers are selected from the group consisting of N,N-disubstituted polyacrylamide and N-substituted polyacrylamide, wherein said N-substituents are 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.
21. The capillary electrophoresis element of claim 20 , wherein the surface interaction component non-crosslinked polymer is poly(N,N-dimethylacrylamide).
22. The capillary electrophoresis element of claim 16 , wherein the composition further comprises at least one denaturant.
23. The capillary electrophoresis element of claim 22 , wherein the at least one denaturant is selected from the group consisting of at least one of formamide, urea, and 2-pyrollidinone.
24. The capillary electrophoresis element of claim 23 , wherein the at least one denaturant comprises urea.
25. The capillary electrophoresis element of claim 16 , wherein the uncoated capillary comprises silica, fused silica, quartz, silicate-based glass, phosphate glass, or alumina-containing glass.
26. The capillary electrophoresis element of claim 16 , wherein the uncoated capillary is a plastic channel capillary.
27. The capillary electrophoresis element of claim 16 , wherein the composition comprises: a sieving component comprising linear polyacrylamide having a molecular weight greater than 1,000,000 Daltons (Da) and less than or equal to about 3,000,000 Da; and having a viscosity of less than 600 centipoise at 25° C.; a surface interaction component comprising poly(N,N-dimethylacrylamide); and a denaturant comprising urea; wherein the composition does not include a crosslinked polymer gel.
28. The capillary electrophoresis element of claim 27 , wherein the uncoated capillary comprises silica, fused silica, quartz, silicate-based glass, such as borosilicate glass, phosphate glass, or alumina-containing glass.
29. The capillary electrophoresis element of claim 28 , wherein the uncoated capillary is a plastic channel capillary.
30. The capillary electrophoresis element of claim 16 , wherein the one or more hydrophobic acrylamide polymers are selected from the group consisting of 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, and —CH 2 CONH 2 , or the N-substituents of the N,N-disubstituted polyacrylamide are taken together with the nitrogen to form a cyclic moiety having a structure of one of the following formulae:
31. The capillary electrophoresis elements of claim 16 , wherein the surface interaction component is present at a concentration in the range of about 0.001% to about 10% w/w.
32. The capillary electrophoresis element of claim 16 , wherein the one or more hydrophobic acrylamide polymers are selected from the group consisting of 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.
33. A kit for separating analytes by capillary electrophoresis comprising a composition comprising: a sieving component comprising an uncrosslinked hydrophilic acrylamide polymer having a molecular weight greater than about 1,000,000 Daltons (Da) and less than or equal to about 3,000,000 Da; and having a viscosity of less than 1000 centipoise; and a surface interaction component comprising one or more non-crosslinked hydrophobic acrylamide polymers wherein the sieving component and the surface interaction component differ in polymer chemical composition; and wherein the composition does not include a crosslinked polymer gel.
34. The kit of claim 33 , wherein the one or more hydrophobic acrylamide polymers are selected from the group consisting of 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, and —CH 2 CONH 2 , or the N-substituents of the N,N-disubstituted polyacrylamide are taken together with the nitrogen to form a cyclic moiety having a structure of one of the following formulae:
35. The kit of claim 33 , wherein the non-crosslinked hydrophilic acrylamide polymer is polyacrylamide.
36. The kit of claim 33 , wherein the one or more hydrophobic acrylamide polymers are selected from the group consisting of 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.
37. A kit for separating analytes by capillary electrophoresis comprising a composition comprising a sieving component comprising an non-crosslinked polyacrylamide polymer having a molecular weight greater than 1,000,000 Daltons (Da) and less than or equal to about 3,000,000 Da; and a viscosity of less than 1000 centipoise at 25° C.;
a surface interaction component comprising one or more non-crosslinked hydrophobic polymers selected from the group consisting of N,N-disubstituted polyacrylamide and N-substituted polyacrylamide, wherein said N-substituents are C 1 to C 3 alkyl;
wherein the sieving component and the surface interaction component differ in polymer chemical composition; and
wherein the composition does not include a crosslinked polymer gel.
38. The kit of claim 37 , wherein the surface interaction component is poly(N,N-dimethylacrylamide).
39. The kit of claim 38 , further comprising at least one denaturant selected from the group consisting of at least one of formamide, urea and 2-pyrollidinone.
40. The kit of claim 39 , wherein the at least one denaturant comprises urea.
41. The kit of claim 33 or 37 , wherein the surface interaction component is present at a concentration in the range of about 0.001% to about 10% w/w.
42. The kit of claim 37 , wherein the non-crosslinked polyacrylamide polymer is linear polyacrylamide.
43. A composition for separating analytes by capillary electrophoresis comprising:
a sieving component comprising non-crosslinked linear polyacrylamide having a molecular weight greater than about 1,000,000 Daltons (Da) and less than or equal to about 3,000,000 Da;
a surface interaction component which is a N,N disubstituted polyacrylamide;
wherein the composition has a viscosity of less than 1,000 centipoise at 25° C. and wherein the composition does not include a crosslinked polymer gel.
44. The composition of any one of claim 1 , 10 , or 43 , wherein the surface interaction component is present at a concentration in the range of about 0.001% to about 10% w/w.