Polymers for separation of biomolecules by capillary electrophoresis
View Patent ↗The invention provides uncharged water-soluble silica-adsorbing polymers for suppressing electroendoosmotic flow and to reduce analyte-wall interactions in capillary electrophoresis. In one aspect of the invention, one or more of such polymers are employed as components of a separation medium for the separation of biomolecules, such as polynucleotides, polysaccharides, proteins, and the like, by capillary electrophoresis. Generally, such polymers are characterized by (i) water solubility over the temperature range between about 20° C. to about 50° C., (ii) concentration in a separation medium in the range between about 0.001% to about 10% (weight/volume), (iii) molecular weight in the range of about 5×10 3 to about 1×10 6 daltons, and (iv) absence of charged groups in an aqueous medium having pH in the range of about 6 to about 9. In one embodiment, polymers of the invention are selected from the group consisting of polylactams, such as polyvinylpyrrolidone; N,N-disubstituted polyacrylamides; and N-substituted polyacrylamides. In accordance with the method of the invention, a sufficient amount of polymer adsorbs to the capillary surface to establish a zone of high viscosity that shields the analyte from the wall and impedes the movement of an electrical double layer under an electric field.
1. A composition for separating polynucleotides by capillary electrophoresis, the composition comprising:
a charge-carrying component;
a surface interaction component comprising one or more polymers 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; and
a sieving component comprises a copolymer,
wherein the composition does not include a crosslinked polymer gel.
2. The composition of claim 1 , wherein said copolymer results from monomeric subunits of at least two of linear polyoxide; polyether; polyacrylamide; polymethacrylamide; polyvinylpyrrolidone; polyvinyloxazolidone; water-soluble hydroxylic polymer; and water-soluble cellulose compound.
3. The composition of claim 1 , wherein said copolymer is used at a concentration of about 0.5% to 10% w.v.
4. The composition of claim 1 , having a viscosity less than 5000 centipoises.
5. The composition of claim 1 , having a viscosity less than 1000 centipoises.
6. The composition of claim 1 , wherein one monomeric subunit of the copolymer is the monomeric subunit of polyacrylamide.
7. The composition of claim 1 , wherein one monomeric subunit of the copolymer is the monomeric subunit of polymethacrylamide.
8. The composition of claim 1 , wherein one monomeric subunit of the copolymer is the monomeric subunit of polyvinylpyrrolidone.
9. A composition for separating polynucleotides by capillary electrophoresis, the composition comprising:
a charge-carrying component;
a surface interaction component comprising one or more uncharged water-soluble silica-adsorbing polymers; and
a sieving component comprising a copolymer, wherein said copolymer results from monomeric subunits of at least two of linear polyoxide; polyether; polyacrylamide; polymethacrylamide; polyvinylpyrrolidone; polyvinyloxazolidone; water-soluble hydroxylic polymer; and water-soluble cellulose compound; wherein the composition does not include a crosslinked polymer gel.
10. The composition of claim 9 , wherein said copolymer is used at a concentration of about 0.5% to 10% w.v.
11. The composition of claim 9 , having a viscosity less than 5000 centipoises.
12. The composition of claim 9 , having a viscosity less than 1000 centipoises.
13. The composition of claim 9 , wherein one monomeric subunit of the copolymer is the monomeric subunit of polyacrylamide.
14. The composition of claim 9 , wherein one monomeric subunit of the copolymer is the monomeric subunit of polymethacrylamide.
15. The composition of claim 9 , wherein one monomeric subunit of the copolymer is the monomeric subunit of polyvinylpyrrolidone.
16. The composition of claim 9 , wherein the surface interaction component comprising one or more uncharged water-soluble silica-adsorbing polymers is non-hydroxylic.
17. The composition of claim 9 , wherein the polyether comprises polyethylene oxide or polypropylene oxide.
18. The composition of claim 9 , wherein the water-soluble hydroxylic polymer comprises xanthan, dextran, or guar.
19. The composition of claim 9 , wherein the water-soluble cellulose compound comprises methylcellulose or hydroxyethylcellulose.