POLYELECTROLYTE-COATED SIZE-EXCLUSION ION-EXCHANGE PARTICLES
A polyelectrolyte-coated particle, devices for using the particle, methods for using the particle for separating PCR reaction products and/or DNA sequencing reaction products, and compositions for coating the particle are provided.
1 . A particle for separating PCR reaction products comprising:
a core comprising ion-exchange material; and
a coating covering the exterior surface of the particle wherein the coating comprises a linear polyelectrolyte polymer, wherein the polyelectrolyte polymer creates a size exclusion barrier allowing small molecules having a size less than 300 bp to penetrate into the particle and restricting large molecules from interacting with the core; and wherein the particle is produced by exposing the core to an excess of the polyelectrolyte polymer.
2 . The particle of claim 1 , wherein the core interacts with at least one PCR reaction product chosen from primers, primer-dimer, ssDNA fragments, unincorporated nucleotides, and salts.
3 . The particle of claim 2 , wherein the particle is adapted to substantially exclude dsDNA fragments having greater than 100 basepairs.
4 .- 5 . (canceled)
6 . The particle of claim 1 , wherein the coating comprises a synthetic polymer having at least one type of charged monomer.
7 . The particle of claim 6 , wherein the synthetic polymer comprises a copolymer, wherein the copolymer comprises at least one monomer chosen from (meth)acrylamide, N-methyl (methyl)acrylamide, N,N-dimethyl (methyl)acrylamide, N-ethyl (meth)acrylamide, N-n-propyl (meth)acrylamide, N-iso-propyl (meth)acrylamide, N-ethyl-N-methyl (meth)acrylamide, N,N-diethyl (meth)acrylamide, N-hydroxymethyl(meth)acrylamide, N-( 3 -hydroxypropyl) (methy)acrylamide, N-vinylformamide, Nvinylacetamide, N-methyl-N-vinylacetamide, vinyl acetate (precursor of vinyl alcohol), 2-hydroxyethyl (meth)acrylate, 3-hydroxypropyl (meth)acrylate, N-vinylpyrrolidone, poly(ethylene oxide) (methy)acrylate, N-(meth)acryloxysuccinimide, N-(meth)acryloylmorpholine, N-2,2,2-trifluoroethyl (meth)acrylamide, N-acetyl (meth)acrylamide, N-amido(meth)acrylamide, N-acetamido (meth)acrylamide, N-tris(hydroxymethyl)methyl (meth)acrylamide, styrenesulfonic acid, N(methyl)acryloyltris(hydroxymethyl)methylamide, (methyl) acryloylurea, vinyloxazolidone, vinylmethyloxazolidone, acrylic acid, methacrylic acid, vinyl sulfonic acid, 4-acetoxystyrene (precursor of 4-hydroxystyrene), vinylphosphonic acid, and vinyl methyl ether.
8 . The particle of claim 7 , wherein the synthetic polymer is poly(acrylic acid-co-N,N-dimethylacrylamide) or poly(N,N-dimethyl acrylamide-co-styrene sulfonic acid).
9 . The particle of claim 1 , wherein the ion-exchange material is porous.
10 . The particle of claim 9 , wherein the ion-exchange material is surface-activated.
11 . The particle of claim 9 , wherein the ion-exchange material has a pore size of 100 Angstroms to 2000 Angstroms.
12 . The particle of claim 11 , wherein the polyelectrolyte polymer has a Mw of 1000 Daltons to 6.0 megaDaltons.
13 . The particle of claim 12 , wherein the ion-exchange material has a pore size of 1000 Angstroms and a Mw of 1.7 megaDaltons to 2.4 megaDaltons.
14 . The particle of claim 6 , wherein the synthetic polymer comprises a copolymer, wherein the copolymer comprises at least one monomer chosen from allyl amide hydrochloride, (3-acrylamidopropyl)trismethylammonium chloride, N-(3aminopropyl) methacrylamide hydrochloride, and N-vinyl amides hydrolyzed to give an amino group.
15 . The particle of claim 14 , wherein the synthetic polymer is poly(N-(3-aminopropyl)methacrylamide-co-N,N-dimethylacrylamide).
16 . The particle of claim 7 , wherein the coating comprises multiple layers.
17 . The particle of claim 16 , wherein the multiple layers comprise polyanions and polycations in alternating layers.
18 . A mixture comprising a cationic ion-exchange material and an anionic ion-exchange material, wherein at least one of the cationic ion-exchange material or the anionic ion-exchange material is a plurality of the particles of claim 1 .
19 . A purification device comprising a receptacle and the mixture of claim 18 disposed in the receptacle.
20 . A microfluidic device comprising a plurality of columns and the mixture of claim 18 disposed in each column.
21 .- 24 . (canceled)
25 . A particle for separating DNA sequencing reaction products, comprising:
a core comprising ion-exchange material; and
a coating covering the entire exterior surface of the particle wherein the coating comprises a linear polyelectrolyte polymer, wherein the polyelectrolyte polymer creates a size exclusion barrier allowing small molecules to penetrate into the particle and substantially excluding dye-labelled ssDNA fragments having greater than 45 nucleotides from interacting with the core; and wherein the particle is produced by exposing the core to an excess of the polyelectrolyte polymer.
26 . The particle of claim 25 , wherein the core interacts with at least one DNA sequencing reaction product chosen from primers, dye-labeled primers, nucleotides, dye-labeled nucleotides, dideoxynucleotides, dye-labeled dideoxynucleotides, and salts.
27 . The particle of claim 26 , wherein the particle is adapted to substantially exclude dye-labeled ssDNA fragments having greater than 10 nucleotides.
28 .- 29 . (canceled)
30 . The particle of claim 25 , wherein the coating comprises a synthetic polymer having at least one type of charged monomer.
31 . The particle of claim 30 , wherein the synthetic polymer comprises a copolymer, wherein the copolymer comprises at least one monomer chosen from (meth)acrylamide, N-methyl (methyl)acrylamide, N,N-dimethyl (methyl)acrylamide, N-ethyl(meth)acrylamide, N-n-propyl (meth)acrylamide, N-iso-propyl (meth)acrylamide, N-ethyl-N-methyl (meth)acrylamide, N,N-diethyl (meth)acrylamide, N-hydroxymethyl(meth)acrylamide, N-(3-hydroxypropyl)(methy)acrylamide, N-vinylformamide, Nvinylacetamide, N-methyl-N-vinylacetamide, vinyl acetate (precursor of vinyl alcohol), 2-hydroxyethyl (meth)acrylate, 3-hydroxypropyl (meth)acrylate, N-vinypyrrolidone, poly(ethylene oxide) (methy)acrylate, N-(meth)acryloxysuccinimide, N-(meth)acryloylmorpholine, N-2,2,2-trifluoroethyl (meth)acrylamide, N-acetyl (meth)acrylamide, N-amido(meth)acrylamide, N-acetamido (meth)acrylamide, N-tris(hydroxymethyl)methyl (meth)acrylamide, styrenesulfonic acid, N(methyl)acryloyltris(hydroxymethyl)methylamide, (methyl) acryloylurea, vinyloxazolidone, vinylmethyloxazolidone, acrylic acid, methacrylic acid, vinyl sulfonic acid, 4-acetoxystyrene (precursor of 4-hydroxystyrene), vinylphosphonic acid, and vinyl methyl ether.
32 . The particle of claim 31 , wherein the synthetic polymer is poly(acrylic acid-co-N,N-dimethylacrylamide) or poly(N,N-dimethyl acrylamide-co-styrene sulfonic acid).
33 . The particle of claim 30 , wherein the synthetic polymer comprises a copolymer, wherein the copolymer comprises at least one monomer chosen from allyl amide hydrochloride, (3-acrylamidopropyl)trismethylammonium chloride, N-(3aminopropyl) methacrylamide hydrochloride, and N-vinyl amides hydrolyzed to give an amino group.
34 . The particle of claim 33 , wherein the synthetic polymer is poly(N-(3-aminopropyl)methacrylamide-co-N,N-dimethylacrylamide).
35 . The particle of claim 25 , wherein the ion-exchange material is porous.
36 . The particle of claim 35 , wherein the ion-exchange material is surface-activated.
37 . The particle of claim 35 , wherein the ion-exchange material has a pore size of 5 Angstrom to 1000 Angstroms.
38 . The particle of claim 37 , wherein the polyelectrolyte polymer has a Mw of 1000 Daltons to 6.0 megaDaltons.
39 . The particle of claim 38 , wherein the ion-exchange material has a pore size of 10 Angstroms to 50 Angstroms and a Mw of 2.4 megaDaltons to 4.9 megaDaltons.
40 . The particle of claim 31 , wherein the polyelectrolyte polymer comprises multiple layers.
41 . The particle of claim 40 , wherein the multiple layers comprise polyanions and polycations in alternating layers.
42 . A mixture comprising a cationic ion-exchange material and an anionic ion-exchange material, wherein at least one of the cationic ion-exchange material or the anionic ion-exchange material is a plurality of the particles of claim 25 .
43 . A purification device comprising a receptacle and the mixture of claim 42 disposed in the receptacle.
44 . A microfluidic device comprising a plurality of columns and the mixture of claim 42 disposed in each column.
45 .- 58 . (canceled)
59 . A system for biological separation, the system comprising:
a linear polyelectrolyte polymer wherein the polyelectrolyte polymer is adapted to coat the exterior surface of ion-exchange material by exposing the ion exchange material to an excess of the polyelectrolyte polymer and to provide sieving for separation of at least one of PCR reaction products or DNA sequencing reaction products wherein the polyelectrolyte polymer creates a size exclusion barrier allowing small molecules having a size less than 300 by to penetrate into the particle and restricting large molecules from interacting with the core.
60 . The system of claim 59 , wherein the system further provides desalting.
61 . The system of claim 59 , wherein the system does not provide desalting.
62 . The system of claim 59 , wherein the ion-exchange material comprises cationic ion-exchange material and anionic ion-exchange material.
63 . The system of claim 62 , wherein the system is in the form of a mixed bed.
64 . The system of claim 63 , wherein the cationic ion-exchange material and the anionic ion-exchange material are present in stoichiometrically equivalent amounts.
65 . (canceled)
66 . The particle of claim 1 , wherein the polyelectrolyte polymer is not crosslinked.
67 . The particle of claim 6 , wherein the charged monomer is at least one type of anionic monomer.
68 . The particle of claim 67 wherein the at least one anionic monomer has a functional group selected from the group consisting of a carboxylic functional group, a boric functional group, a sulfonic functional group, a sulfinic functional group, a phosphoric functional group, a phosphorus functional group, a phenolic functional group, or a combination thereof.
69 . The particle of claim 6 , wherein the polyelectrolyte synthetic polymer is a homopolymer.
70 . The particle of claim 69 , wherein the homopolymer is selected from the group consisting of styrene sulfonic acid, acrylic acid, methacrylic acid, vinyl sulfonic acid, vinyl phosphonic acid, allyl amine hydrochloride, (3-acrylamidopropyl) trimethylammonium chloride, N-(3-aminopropyl) methacrylamide hydrochloride, fumaric acid, aspartic acid, galactronic acid, glutamic acid, adenylic acid, inosinic acid, and uridylic acid.
71 . The mixture of claim 18 , wherein the anionic ion-exchange material is a plurality of the particles of claim 1 .
72 . The particle of claim 25 , wherein the polyelectrolyte polymer is not crosslinked.
73 . The particle of claim 30 , wherein the charged monomer is at least one type of anionic monomer.
74 . The particle of claim 73 wherein the at least one anionic monomer has a functional group selected from the group consisting of a carboxylic functional group, a boric functional group, a sulfonic functional group, a sulfinic functional group, a phosphoric functional group, a phosphorus functional group, a phenolic functional group, or a combination thereof.
75 . The particle of claim 30 , wherein the polyelectrolyte synthetic polymer is a homopolymer.
76 . The particle of claim 75 , wherein the homopolymer is selected from the group consisting of styrene sulfonic acid, acrylic acid, methacrylic acid, vinyl sulfonic acid, vinyl phosphonic acid, allyl amine hydrochloride, (3-acrylamidopropyl) trimethylammonium chloride, N-(3-aminopropylmethacrylamide hydrochloride, fumaric acid, aspartic acid, galactronic acid, glutamic acid, adenylic acid, inosinic acid, and uridylic acid.