High speed, high resolution compositions, methods and kits for capillary electrophoresis
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 method for separating analytes by capillary electrophoresis comprising:
placing the analytes in a composition comprising:
a sieving component comprising a non-crosslinked acrylamide polymer having a weight average molecular weight between about 3,000,000 Daltons (Da) Da and less than or equal to about 6,400,000 Da; and
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 the 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 are the same or different; and wherein the composition does not include a cross-linked polymer gel; 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:
separating the analytes by capillary electrophoresis with 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 acrylamide polymer having a weight average molecular weight between about 3,000,000 Daltons (Da) and less than or equal to about 6,400,000 Da; and a surface interaction component;
wherein the surface interaction component comprises a solution of one or more non-crosslinked polymers; and
wherein the capillary electrophoresis element does not include a crosslinked polymeric gel.
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 comprise a plastic substrate.
10. 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 sieving component comprising an uncrosslinked acrylamide polymer having a weight average molecular weight between about 3,000,000 Da and less than or equal to about 6,400,000 Da and 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-substitute polyacrylamide, wherein the 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 are the same or different; wherein the compositions does not include a crosslinked polymer gel;
loading a sample of different sized analytes into the capillary; and
applying an electrical field between the first and second ends of the capillary so that the different sized analytes in the sample migrate through the capillary, thereby separating the analytes.
11. The method of claim 10 , wherein the composition further comprises at least one denaturant.
12. The method of claim 11 , wherein the at least one denaturant comprises urea.
13. The method of claim 10 , wherein the surface interaction non-cross-linked polymer is poly(N,N-dimethylacrylamide).
14. The method of claim 10 , which is carried out in parallel with a plurality of uncoated capillaries.
15. The method of claim 14 , wherein the uncoated capillaries comprise silica, fused silica, quartz, silicate-based glass, phosphate glass, or alumina-containing glass.
16. The method of claim 14 , wherein the uncoated capillaries comprise a plastic substrate.