Poly and copoly(
The invention relates generally to polymers and copolymers comprising N-vinylamide-type monomers, their preparation, and compositions, such as electrophoresis separation media, containing the same; to supports, such as capillaries, containing these polymers; and methods for separating a mixture of biomolecules, especially polynucleotides, using capillary electrophoresis. Separation media comprising such polymers yield advantageous performance in the analysis and separation of biomolecules by capillary electrophoresis.
1. A composition comprising a buffer, a denaturant, an effective amount of a sieve and/or EOF suppressing polymer comprising a monomer unit, and a uncrosslinked water soluble polymer having the form poly(M 1 x M 2 y ), or a salt thereof, comprising one or more monomers of type M 1 , wherein:
(a) y is zero;
(b) each monomer in the polymer is of type M 1 ;
(c) x is an integer ranging from 1 to 5 and represents the number of monomer subtypes of type M 1 that are present in the polymer; and
(d) each monomer subtype of type M 1 in the polymer is N-vinylhexamide, N-vinylheptamide, N-vinyloctamide, N-vinylnonamide, N-vinyldecamide, N-vinylformamide, N-methyl-N-vinylformamide, N-vinylacetamide, N-methyl-N-vinylacetamide, N-methyl-N-vinylpropamide, N-methyl-N-vinylbutamide, N-methyl-N-vinylpentamide, N-methyl-N-vinylhexamide, N-methyl-N-vinylheptamide, N-methyl-N-vinyloctamide, N-methyl-N-vinylnonamide, N-methyl-N-vinyldecamide, N-ethyl-N-vinylformamide, N-ethyl-N-vinylacetamide, N-ethyl-N-vinylpropamide, N-ethyl-N-vinylbutamide, N-ethyl-N-vinylpentamide, N-ethyl-N-vinylhexamide, N-ethyl-N-vinylheptamide, N-ethyl-N-vinyloctamide, N-ethyl-N-vinylnonamide, N-ethyl-N-vinyldecamide, N-n-propyl-N-vinylformamide, N-n-propyl-N-vinylacetamide, N-n-propyl-N-vinylpropamide, N-n-propyl-N-vinylbutamide, N-n-propyl-N-vinylpentamide, N-n-propyl-N-vinylhexamide, N-n-propyl-N-vinylheptamide, N-n-propyl-N-vinyloctamide, N-n-propyl-N-vinylnonamide, N-n-propyl-N-vinyldecamide, N-iso-propyl-N-vinylformamide, N-iso-propyl-N-vinylacetamide, N-iso-propyl-N-vinylpropamide, N-iso-propyl-N-vinylbutamide, N-iso-propyl-N-vinylpentamide, N-iso-propyl-N-vinylhexamide, N-iso-propyl-N-vinylheptamide, N-iso-propyl-N-vinyloctamide, N-iso-propyl-N-vinylnonamide or N-iso-propyl-N-vinyldecamide, and
wherein the sieve and/or EOF suppressing polymer monomer unit is acrylamide, N-acetyl-acrylamide, N-2-cyanoethyl-acrylamide, N,N-1,2-dihydroxyethylene-bis-acrylamide, N-4,4-dimethoxybutyl-acrylamide, N-2,2-dimethoxyethyl-acrylamide, N,N-dimethyl-acrylamide, N-2-glycolic acid methyl ester acrylamide, N-2-hydroxyethyl-acrylamide, N-hydroxymethyl-acrylamide, N-methoxymethyl-acrylamide, N-3-methoxypropyl-acrylamide, N-methyl-acrylamide, N-methyl-, N-2,2-dimethoxyethyl-acrylamide, N-morpholinoethyl-acrylamide, N-2,2,2-trichloro-1-hydroxyethyl-acrylamide, N-tri(hydroxymethyl)-methyl-acrylamide, methacrylamide, N-acetyl-methacrylamide, N-2-cyanoethyl-methacrylamide, N,N-1,2-dihydroxyethylene-bis-methacrylamide, N-4,4-dimethoxybutyl-methacrylamide, N-2,2-dimethoxyethyl-methacrylamide, N,N-dimethyl-methacrylamide, N-2-glycolic acid methyl ester methacrylamide, N-2-hydroxyethyl-methacrylamide, N-hydroxymethyl-methacrylamide, N-methoxymethyl-methacrylamide, N-3-methoxypropyl-methacrylamide, N-methyl-methacrylamide, N-methyl-, N-2,2-dimethoxyethyl-methacrylamide, N-morpholinoethyl-methacrylamide, N-2,2,2-trichloro-1-hydroxyethyl-methacrylamide, N-tri(hydroxymethyl)-methyl-methacrylamide, or a mixture thereof.
2. The composition of claim 1 , wherein the sieve and/or EOF suppressing polymer monomer unit is acrylamide.
3. The composition of claim 1 , wherein the polymer has a weight-average molecular weight of from about 150,000 Daltons to about 20 MDaltons.
4. The composition of claim 1 , wherein the composition is an aqueous composition.
5. The composition of claim 4 , wherein the composition has a pH of from about 5 to about 11.
6. The composition of claim 5 , wherein the denaturant is formamide, urea, pyrrolidone, N-methyl pyrrolidone, or a mixture thereof.
7. The composition of claim 6 , wherein the denaturant is urea.
8. The composition of claim 6 , wherein the denaturant is formamide.
9. The composition of claim 4 , wherein the composition has a pH of from about 7 to about 10.
10. The composition of claim 4 , wherein y is 0 and each M 1 is independently N-vinylformamide or N-methyl-N-vinylacetamide.
11. A capillary containing the composition of claim 1 .
12. The capillary of claim 11 , wherein the capillary is a capillary tube.
13. The composition of claim 1 , wherein the Mw of the polymer is from about 500,000 Daltons to about 5MDaltons.
14. The composition of claim 1 wherein the polymer is a homopolymer.
15. A method for separating a mixture of biomolecules, comprising:
(a) contacting a composition with a mixture comprising a biomolecule; and
(b) applying an electric field to the composition in an amount sufficient to facilitate the separation of a biomolecule from the mixture,
wherein the composition comprises a buffer, a denaturant, and a uncrosslinked water soluble polymer having the form poly(M 1 x M 2 y ), or a salt thereof, comprising one or more monomers of type M 1 , wherein:
(i) y is zero;
(ii) each monomer in the polymer is of type M 1 ;
(iii) x is an integer ranging from 1 to 5 and represents the number of monomer subtypes of type M 1 that are present in the polymer; and
(iv) each monomer subtype of type M 1 in the polymer is N-vinylhexamide, N-vinylheptamide, N-vinyloctamide, N-vinylnonamide, N-vinyldecamide, N-vinylformamide, N-methyl-N-vinylformamide, N-vinylacetamide, N-methyl-N-vinylacetamide, N-methyl-N-vinylpropamide, N-methyl-N-vinylbutamide, N-methyl-N-vinylpentamide, N-methyl-N-vinylhexamide, N-methyl-N-vinylheptamide, N-methyl-N-vinyloctamide, N-methyl-N-vinylnonamide, N-methyl-N-vinyldecamide, N-ethyl-N-vinylformamide, N-ethyl-N-vinylacetamide, N-ethyl-N-vinylpropamide, N-ethyl-N-vinylbutamide, N-ethyl-N-vinylpentamide, N-ethyl-N-vinylhexamide, N-ethyl-N-vinylheptamide, N-ethyl-N-vinyloctamide, N-ethyl-N-vinylnonamide, N-ethyl-N-vinyldecamide, N-n-propyl-N-vinylformamide, N-n-propyl-N-vinylacetamide, N-n-propyl-N-vinylpropamide, N-n-propyl-N-vinylbutamide, N-n-propyl-N-vinylpentamide, N-n-propyl-N-vinylhexamide, N-n-propyl-N-vinylheptamide, N-n-propyl-N-vinyloctamide, N-n-propyl-N-vinylnonamide, N-n-propyl-N-vinyldecamide, N-iso-propyl-N-vinylformamide, N-iso-propyl-N-vinylacetamide, N-iso-propyl-N-vinylpropamide, N-iso-propyl-N-vinylbutamide, N-iso-propyl-N-vinylpentamide, N-iso-propyl-N-vinylhexamide, N-iso-propyl-N-vinylheptamide, N-iso-propyl-N-vinyloctamide, N-iso-propyl-N-vinylnonamide or N-iso-propyl-N-vinyldecamide.
16. The method of claim 15 , wherein the separation is performed within a capillary tube and two or more biomolecules are polynucleotides.
17. The method of claim 16 , wherein the method for separating has a crossover of at least about 400 base pairs.
18. A composition comprising a buffer, a denaturant, an effective amount of a sieve and/or EOF suppressing polymer comprising a monomer unit, and a uncrosslinked water soluble polymer having the form poly(M 1 x M 2 y ), or a salt thereof, comprising one or more monomers of type M 1 wherein:
(a) y is zero;
(b) each monomer in the polymer is of type M 1 ;
(c) x is an integer ranging from 1 to 5 and represents the number of monomer subtypes of type M 1 that are present in the polymer; and
(d) each monomer subtype of type M 1 in the polymer is N-vinylhexamide, N-vinylheptamide, N-vinyloctamide, N-vinylnonamide, N-vinyldecamide, N-vinylformamide, N-methyl-N-vinylformamide, N-vinylacetamide, N-methyl-N-vinylacetamide, N-methyl-N-vinylpropamide, N-methyl-N-vinylbutamide, N-methyl-N-vinylpentamide, N-methyl-N-vinylhexamide, N-methyl-N-vinylheptamide, N-methyl-N-vinyloctamide, N-methyl-N-vinylnonamide, N-methyl-N-vinyldecamide, N-ethyl-N-vinylformamide, N-ethyl-N-vinylacetamide, N-ethyl-N-vinylpropamide, N-ethyl-N-vinylbutamide, N-ethyl-N-vinylpentamide, N-ethyl-N-vinylhexamide, N-ethyl-N-vinylheptamide, N-ethyl-N-vinyloctamide, N-ethyl-N-vinylnonamide, N-ethyl-N-vinyldecamide, N-n-propyl-N-vinylformamide, N-n-propyl-N-vinylacetamide, N-n-propyl-N-vinylpropamide, N-n-propyl-N-vinylbutamide, N-n-propyl-N-vinylpentamide, N-n-propyl-N-vinylhexamide, N-n-propyl-N-vinylheptamide, N-n-propyl-N-vinyloctamide, N-n-propyl-N-vinylnonamide, N-n-propyl-N-vinyldecamide, N-iso-propyl-N-vinylformamide, N-iso-propyl-N-vinylacetamide, N-iso-propyl-N-vinylpropamide, N-iso-propyl-N-vinylbutamide, N-iso-propyl-N-vinylpentamide, N-iso-propyl-N-vinylhexamide, N-iso-propyl-N-vinylheptamide, N-iso-propyl-N-vinyloctamide, N-iso-propyl-N-vinylnonamide or N-iso-propyl-N-vinyldecamide, and
wherein the sieve and/or EOF suppressing polymer has having a molecular weight of from about 100,000 Daltons to about 5 MDaltons.
19. The composition of claim 18 , wherein the sieve and/or EOF suppressing polymer comprises or is poly(hydroxymethylene), poly(oxyethylene), poly(oxypropylene), poly(oxyethylene-co-oxypropylene), poly(vinyl alcohol), poly(vinylpyrrolidone), poly(2-ethyl-2-oxazoline), poly(2-methyl-2oxazoline), poly ((2-ethyl-2-oxazoline)-co-(2-methyl-2-oxazoline)), poly(N-acetamidoacrylamide), poly(acryloxylurea), hydroxyethyl cellulose, hydroxymethyl cellulose, or a mixture thereof.
20. The composition of claim 18 , wherein the sieve and/or EOF suppressing polymer has a weight-average molecular weight of from about 1 MDalton to about 5 MDaltons.
21. The composition of claim 20 , wherein the sieve and/or EOF suppressing polymer is substantially linear polyacrylamide.