CN101081373A - Cation-exchange crystal glue chromatography medium and method for preparing the same - Google Patents
Cation-exchange crystal glue chromatography medium and method for preparing the same Download PDFInfo
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- CN101081373A CN101081373A CNA2007100698189A CN200710069818A CN101081373A CN 101081373 A CN101081373 A CN 101081373A CN A2007100698189 A CNA2007100698189 A CN A2007100698189A CN 200710069818 A CN200710069818 A CN 200710069818A CN 101081373 A CN101081373 A CN 101081373A
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- 239000013078 crystal Substances 0.000 title claims abstract description 50
- 238000005341 cation exchange Methods 0.000 title claims description 29
- 238000000034 method Methods 0.000 title claims description 4
- 239000003292 glue Substances 0.000 title description 12
- 239000012501 chromatography medium Substances 0.000 title description 3
- 239000000178 monomer Substances 0.000 claims abstract description 37
- 239000011159 matrix material Substances 0.000 claims abstract description 34
- 238000001816 cooling Methods 0.000 claims abstract description 25
- 239000003054 catalyst Substances 0.000 claims abstract description 21
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims abstract description 17
- 238000006243 chemical reaction Methods 0.000 claims abstract description 17
- 238000002360 preparation method Methods 0.000 claims abstract description 14
- 239000003431 cross linking reagent Substances 0.000 claims abstract description 13
- 239000000495 cryogel Substances 0.000 claims abstract description 11
- 238000010438 heat treatment Methods 0.000 claims abstract description 4
- 239000002609 medium Substances 0.000 claims description 55
- 239000000499 gel Substances 0.000 claims description 47
- 239000000243 solution Substances 0.000 claims description 21
- ROOXNKNUYICQNP-UHFFFAOYSA-N ammonium persulfate Chemical compound [NH4+].[NH4+].[O-]S(=O)(=O)OOS([O-])(=O)=O ROOXNKNUYICQNP-UHFFFAOYSA-N 0.000 claims description 16
- 239000000084 colloidal system Substances 0.000 claims description 13
- 229920000642 polymer Polymers 0.000 claims description 13
- 238000002425 crystallisation Methods 0.000 claims description 11
- KWYHDKDOAIKMQN-UHFFFAOYSA-N N,N,N',N'-tetramethylethylenediamine Chemical compound CN(C)CCN(C)C KWYHDKDOAIKMQN-UHFFFAOYSA-N 0.000 claims description 8
- 229910001870 ammonium persulfate Inorganic materials 0.000 claims description 8
- 230000000694 effects Effects 0.000 claims description 8
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 claims description 7
- 125000000524 functional group Chemical group 0.000 claims description 7
- 150000002500 ions Chemical class 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 239000000376 reactant Substances 0.000 claims description 7
- 238000010792 warming Methods 0.000 claims description 7
- LVHBHZANLOWSRM-UHFFFAOYSA-N methylenebutanedioic acid Natural products OC(=O)CC(=C)C(O)=O LVHBHZANLOWSRM-UHFFFAOYSA-N 0.000 claims description 6
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims description 5
- 230000035484 reaction time Effects 0.000 claims description 5
- JAHNSTQSQJOJLO-UHFFFAOYSA-N 2-(3-fluorophenyl)-1h-imidazole Chemical compound FC1=CC=CC(C=2NC=CN=2)=C1 JAHNSTQSQJOJLO-UHFFFAOYSA-N 0.000 claims description 4
- JKNCOURZONDCGV-UHFFFAOYSA-N 2-(dimethylamino)ethyl 2-methylprop-2-enoate Chemical compound CN(C)CCOC(=O)C(C)=C JKNCOURZONDCGV-UHFFFAOYSA-N 0.000 claims description 4
- 125000003368 amide group Chemical group 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 4
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 claims description 3
- 230000008025 crystallization Effects 0.000 claims description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 3
- ZIUHHBKFKCYYJD-UHFFFAOYSA-N n,n'-methylenebisacrylamide Chemical compound C=CC(=O)NCNC(=O)C=C ZIUHHBKFKCYYJD-UHFFFAOYSA-N 0.000 claims description 2
- 239000011148 porous material Substances 0.000 abstract description 12
- 238000001179 sorption measurement Methods 0.000 abstract description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 11
- 125000002091 cationic group Chemical group 0.000 abstract 3
- -1 crystallizing Substances 0.000 abstract 1
- 230000000379 polymerizing effect Effects 0.000 abstract 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 238000004587 chromatography analysis Methods 0.000 description 8
- 102000016943 Muramidase Human genes 0.000 description 7
- 108010014251 Muramidase Proteins 0.000 description 7
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 7
- 229960000274 lysozyme Drugs 0.000 description 7
- 235000010335 lysozyme Nutrition 0.000 description 7
- 239000004325 lysozyme Substances 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 230000008929 regeneration Effects 0.000 description 5
- 238000011069 regeneration method Methods 0.000 description 5
- 239000008367 deionised water Substances 0.000 description 4
- 229910021641 deionized water Inorganic materials 0.000 description 4
- 239000011521 glass Substances 0.000 description 4
- 238000010559 graft polymerization reaction Methods 0.000 description 4
- 238000007789 sealing Methods 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 2
- 239000005977 Ethylene Substances 0.000 description 2
- IZZSMHVWMGGQGU-UHFFFAOYSA-L disodium;2-methylidenebutanedioate Chemical compound [Na+].[Na+].[O-]C(=O)CC(=C)C([O-])=O IZZSMHVWMGGQGU-UHFFFAOYSA-L 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- SMZOUWXMTYCWNB-UHFFFAOYSA-N 2-(2-methoxy-5-methylphenyl)ethanamine Chemical compound COC1=CC=C(C)C=C1CCN SMZOUWXMTYCWNB-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N 2-Propenoic acid Natural products OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- NRTTULQAGVYCSY-UHFFFAOYSA-N 2-methylidenebutanedioic acid;potassium Chemical compound [K].OC(=O)CC(=C)C(O)=O NRTTULQAGVYCSY-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 125000006297 carbonyl amino group Chemical group [H]N([*:2])C([*:1])=O 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
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Abstract
The present invention discloses one new type of cationic exchange macroporous chromatographic cryogel medium for continuous bed and its preparation process. The cationic exchange cryogel medium has functional carboxyl group, pore size of 5-400 micron, and porosity of 50-98 %. It is prepared through adding catalyst into water solution of cryogel monomer and cross-linking agent, crystallizing, pore creating and polymerizing reaction in a cooling system, heating to thaw ice crystal so as to form very large pores and obtain cryogel matrix, and fixing monomer with carboxyl group and capable of producing grafting reaction with the cryogel matrix onto the cryogel matrix at 30-90 deg.c and under the action of catalyst to obtain the cationic exchange cryogel medium. The cryogel medium has good connectivity, excellent separating performance and great adsorption capacity, and may be regenerated easily for reuse to over 20 times.
Description
(1) technical field
The invention belongs to bio-separation and medical technical field, relate to a kind of cation exchange type super macroporous continous bed crystal gel (Cryogel) chromatography medium and preparation method thereof.
(2) background technology
Super macroporous continuous bed crystalloid colloid chromatographic separating process (Supermacroporous CryogelChromatography, be called for short crystal glue chromatography), be a kind of new bio chromatography technology that occurred in 2002, can under high flow rate, realize from material liquids such as complicated microbial fermentation solution, nutrient solution, lysate, directly extracting and the separate targets thing.The brilliant glue of ion-exchange is the medium that often uses in the crystal glue chromatography, and very effective aspect the absorption of large biological molecule and chromatography, bigger to the macromolecular adsorption capacity of target organism, wash-out is convenient.Therefore, obtained extensive use in the separation and purification field of large biological molecule and drug molecule.
With carboxyl (is that the brilliant glue of functional group is a kind of in the cation exchange crystal gel medium COO-).In the existing documents and materials (Savina etc., Polymer 46,9596-9603,2005) being the glycerol polymerization monomer with acrylic acid once, is the medium skeleton with the brilliant gel matrix of polyacrylamide base, by glycerol polymerization, obtained a kind of cation exchange crystal gel medium, its cation exchange functional group is
Handle but need that in advance matrix is carried out drying, carry out graft reaction then, the adsorption capacity of gained medium need improve.
(3) summary of the invention
The purpose of this invention is to provide a kind of new cation exchange type super macroporous continous bed crystal gel separating medium that has carboxyl type functional group.
Cation exchange super macroporous continuous bed crystalloid colloid medium of the present invention (cryogel) aperture is 5～400 μ m, porosity 50～98%, and described crystal gel medium has following carboxyl type cation exchange functional group:
Wherein said functional group is by brilliant gel matrix and can forms the group that has in the polymer with the monomer reaction that has carboxyl of brilliant gel matrix generation graft reaction.
Described crystal gel medium has the less height equivalent to one theoretical plate (HETP) and bigger adsorption capacity, and when flow velocity during at 0.1～10cm/min, the brilliant glue column height equivalent to one theoretical plate (HETP) is less than 0.2cm, and adsorption capacity is greater than 10mg lysozyme/g crystal gel medium.
The invention provides a kind of preparation method of described cation exchange type super macroporous continous bed crystal gel medium, comprise the steps:
(1) bed skeleton polymer monomer and crosslinking agent are prepared into the aqueous solution of gross mass percent concentration 2～14%, after adding catalyst A reactant liquor is placed the cooling system crystallization and carries out polymerisation, heating up and make crystal form the super large hole in the back, obtains described super macroporous continuous bed crystalloid colloid matrix; Described bed skeleton polymer monomer is the polymerisable monomer that contains amino or amide groups; Described A catalyst is one of following: mixture, the triethanolamine of ammonium persulfate and tetramethylethylenediamine arbitrary proportion; Described polymer monomer: crosslinking agent: the mass ratio that feeds intake of catalyst A is 1: 0.01～0.5: 0.001～0.1.
(2) can with the monomer that has carboxyl of brilliant gel matrix generation graft reaction under the effect of catalyst B, immobilized in crystal gel medium matrix by graft reaction, promptly obtain described cation exchange crystal gel medium, described catalyst B is a high price Cu ion solution.
The monomer that has carboxyl described in the above-mentioned steps (2) is made into the solution that concentration is 0.001～5mol/L earlier when using, and the described consumption that has the monomer solution of carboxyl is 0.5～10 times of described crystal gel medium matrix volume.
Further, the described monomer that has a carboxyl can with matrix generation graft reaction, the optional monomer that ethylene linkage and carboxyl are arranged, the combination of one of preferred following or two kinds of arbitrary proportions: itaconic acid, itaconate.Described ethylene linkage (C=C) can with the amido link (CONH in the crystal gel medium matrix
2-) under the effect of catalyst B, carry out graft reaction, thus functional group is immobilized in brilliant gel matrix.
Catalyst B can be the high price Cu ion solution of 0.001～0.5mol/L in the described step (2), and the volumetric usage of described high price Cu ion solution is more than 0.5 times of described brilliant gel matrix volume.
The temperature of graft reaction is at 30～90 ℃ in the step (2), preferred 40～60 ℃; 0.5～24 hour reaction time, preferred 1～5 hour.
Described bed skeleton polymer monomer is the polymerisable monomer that contains amino or amide groups, can be the mixture of following one or both or two or more arbitrary proportions: acrylamide (AAm), N, N-DMAA (DMAAm), dimethylaminoethyl methacrylate (DMAEMA), more preferably acrylamide (AAm) or N,N-DMAA (DMAAm).
Described crosslinking agent is one of following: N, and N '-methylene-bisacrylamide (MBAAm), N, N '-diene propiono ethylenediamine is preferably N, N '-methylene-bisacrylamide (MBAAm).
Described catalyst A will be selected at different polymer monomers, normal is corresponding initiator of polyreaction or accelerator, at the present invention concrete bed skeleton polymer monomer and the crosslinking agent that uses, it is one of following that the A catalyst can be: mixture, the triethanolamine of ammonium persulfate and tetramethylethylenediamine arbitrary proportion; Be preferably the mixture of ammonium persulfate (APS) and tetramethylethylenediamine (TEMED).
Reactant liquor crystallisation by cooling under-60～0 ℃ of conditions in cooling system in the described step (1).
Further, the thermal history of reactant liquor crystallisation by cooling system is in the described step (1):
(A) cooling: drop to-30～-10 ℃ by 0 ℃;
(B) constant temperature: constant temperature 5～24 hours;
(C) heat up: be warming up to room temperature, at room temperature make crystal form the super large hole, obtain described super macroporous continuous bed crystalloid colloid matrix.
Further, temperature-fall period can carry out step by step, can drop to one earlier than higher temperature, and constant temperature cools to the temperature of reduction again after a period of time, progressively carry out, up to the temperature that drops to setting.
Crystal gel medium with method preparation provided by the invention has following characteristic:
1) physical property of crystal gel medium: porosity 50～98%, pore diameter range 5～400 μ m are connective good.With water is example, when water flow velocity in 0.1～10cm/min scope, the structure of medium is constant substantially.
2) adsorption separation performance of crystal gel medium: flow velocity is in 0.1～10cm/min scope, and the brilliant glue column height equivalent to one theoretical plate (HETP) is less than 0.2cm, and separating property is good, is example with the lysozyme, and adsorption capacity is greater than 10mg lysozyme/g medium.
3) life-span of crystal gel medium: can regenerate easily, reuse number of times and reach more than 20 times.
Cation exchange type crystal gel medium of the present invention and preparation thereof have following advantage:
1) the crystal gel medium resistance to mass tranfer is little, and is connective good, can operate in the high flow rate scope;
2) adsorption capacity to large biological molecule is bigger, the separative efficiency height, and wash-out is easy, and regeneration is convenient;
3) application is wide: allow microbial cell or cell fragment to pass through smoothly, can be under high flow rate from complicated feed liquid such as the zymotic fluid that contains microbial cell, cell fragment etc., nutrient solution, lysate system direct separate targets thing, be suitable for the extensive separation and Extraction and the purifying of genetic engineering downstream targets thing, conventional fermentate, biochemical drug etc.
4) monomer material is easy to get, and does not need brilliant gel matrix dryly in advance in the graft reaction, and the graft reaction mild condition is easily gone, and preparation technology is simple, and cost is low, and large-scale production is very easy.
(4) specific embodiment:
Below with specific embodiment technical scheme of the present invention is described, but protection scope of the present invention is not limited thereto:
Embodiment 1
With 2.25g DMAEMA and AAm (mass ratio 0.5: 1) monomer, 0.0225g crosslinking agent MBAAm is dissolved in the 14ml deionized water, after stirring, add 10.5mgTEMED and 12mg APS rapidly, the gained mixed liquor is packed in the glass chromatography column of internal diameter 16mm, long 100mm, after the sealing, but in the constant temperature cooling system of temperature programmed control, carry out the crystallisation by cooling pore.Thermal history is:
(A) cooling: drop to-30 ℃ by 0 ℃;
(B) constant temperature: constant temperature 24 hours;
(C) heat up: be warming up to room temperature.
Then, at room temperature melt crystal, obtain super macroporous continuous bed crystalloid colloid medium matrix, volume 16ml.With the sodium itaconate monomer solution of matrix and 160ml concentration 0.001mol/L, at 75ml 0.005mol/L Cu
3+The solution catalyzing effect under, under 30 ℃, carry out graft polymerization reaction 24h, obtain the cation exchange crystal gel medium.Its porosity 50%, pore diameter range 5～80 μ m; Water flow velocity is in 0.1～10cm/min scope, and the structure of medium is constant, height equivalent to one theoretical plate (HETP) 0.06cm in the brilliant glue column; Adsorption capacity 12mg lysozyme/g medium with 2mol/L NaCl and 0.5mol/L NaOH regeneration, is reused more than 30 times.
Embodiment 2
With 2.77g DMAAm monomer; 0.554g crosslinking agent N; N '-diene propiono ethylenediamine is dissolved in the 53ml deionized water; after stirring; add rapidly the 14mg triethanolamine, the gained mixed liquor is packed in the glass chromatography column of internal diameter 26mm, long 150mm, after the sealing; but in the constant temperature cooling system of temperature programmed control, carry out the crystallisation by cooling pore.Thermal history is:
(A) cooling: drop to-60 ℃ by 0 ℃;
(B) heat up: be warming up to-4 ℃;
(C) constant temperature: constant temperature 1 hour;
(D) cooling: be cooled to-15 ℃ again by-4 ℃;
(E) constant temperature: constant temperature 18 hours;
(F) heat up: be warming up to room temperature.
Then, at room temperature melt crystal, form the super large hole, obtain super macroporous continuous bed crystalloid colloid medium matrix, volume 56ml.With the sodium itaconate of matrix and 120ml 2.5mol/L and the monomer solution (1: 1) of 2.5mol/L itaconic acid, at 150ml 0.1mol/L Cu
3+The solution catalyzing effect under, under 60 ℃, carry out graft polymerization reaction time 5h, obtain the cation exchange crystal gel medium.Its porosity 86%, pore diameter range 20～250 μ m; Water flow velocity is in 0.1～10cm/min scope, and the structure of medium is constant, height equivalent to one theoretical plate (HETP) 0.13cm in the brilliant glue column; Adsorption capacity 36mg lysozyme/g medium with 1mol/L NaCl and 0.2mol/L NaOH regeneration, is reused more than 25 times.
Embodiment 3
With 0.76gAAm and DMAAm (1: 0.2) monomer, 0.38g crosslinking agent MBAAm is dissolved in the 12ml deionized water, after stirring, add 40mg TEMED and 36mgAPS rapidly, the gained mixed liquor is packed in the glass chromatography column of internal diameter 10mm, long 100mm, after the sealing, but in the constant temperature cooling system of temperature programmed control, carry out the crystallisation by cooling pore.Thermal history is:
(A) cooling: drop to-22 ℃ by 0 ℃;
(B) constant temperature: constant temperature 8 hours;
(C) heat up: be warming up to room temperature.
Then, at room temperature melt crystal, obtain super macroporous continuous bed crystalloid colloid medium matrix, volume 13ml.With the itaconic acid potassium monomer solution of matrix and 52ml 0.5mol/L, at 6.51ml0.5mol/L Cu
3+The solution catalyzing effect under, under 90 ℃, carry out graft polymerization reaction time 0.5h, obtain the cation exchange crystal gel medium.Its porosity 73%, pore diameter range 5～150 μ m; Water flow velocity is in 0.1～10cm/min scope, and the structure of medium is constant, height equivalent to one theoretical plate (HETP) 0.08cm in the brilliant glue column; Adsorption capacity 52mg lysozyme/g medium with 2mol/L NaCl and 0.5mol/L NaOH regeneration, is reused more than 25 times.
Embodiment 4
With 1.2g AAm monomer, 0.1g crosslinking agent MBAAm is dissolved in the 63.5ml deionized water, after stirring, add 10mg TEMED and 14mg APS rapidly, the gained mixed liquor is packed in the glass chromatography column of internal diameter 26mm, long 150mm, after the sealing, but in the constant temperature cooling system of temperature programmed control, carry out the crystallisation by cooling pore.Thermal history is:
(A) cooling: drop to-10 ℃ by 0 ℃;
(B) constant temperature: constant temperature 18 hours;
(C) heat up: be warming up to room temperature.
Then, at room temperature melt crystal, obtain super macroporous continuous bed crystalloid colloid medium matrix, volume 64ml.With the itaconic acid monomer solution of matrix and 65ml concentration 1.8mol/L, at the Cu of 100ml0.3mol/L
3+The solution catalyzing effect under, under 45 ℃, carry out graft polymerization reaction time 2h, obtain the cation exchange crystal gel medium.Its porosity 98%, pore diameter range 50～400 μ m; Water flow velocity is in 0.1～10cm/min scope, and the structure of medium is constant, height equivalent to one theoretical plate (HETP) 0.197cm in the brilliant glue column; Adsorption capacity 111mg lysozyme/g medium, with 1mol/L NaCl and 0.5mol/L NaOH regeneration, reusable more than 25 times.
Claims (10)
1. a cation exchange super macroporous continuous bed crystalloid colloid medium (cryogel) is characterized in that described crystal gel medium aperture is 5～400 μ m, porosity 50～98%, and described crystal gel medium has following carboxyl type cation exchange functional group:
The preparation of described cation exchange super macroporous continuous bed crystalloid colloid medium comprises the steps:
(1) bed skeleton polymer monomer and crosslinking agent are prepared into the aqueous solution of gross mass percent concentration 2～14%, after adding catalyst A reactant liquor is placed the cooling system crystallization and carries out polymerisation, heating up and make crystal form the super large hole in the back, obtains described super macroporous continuous bed crystalloid colloid matrix; Described catalyst A is one of following: mixture, the triethanolamine of ammonium persulfate and tetramethylethylenediamine arbitrary proportion;
(2) can with the monomer that has carboxyl of brilliant gel matrix generation graft reaction under the effect of catalyst B, immobilized in crystal gel medium matrix by graft reaction, promptly obtain described cation exchange crystal gel medium; Described catalyst B is a high price Cu ion solution.
2. a method for preparing cation exchange type super macroporous continous bed crystal gel medium as claimed in claim 1 comprises the steps:
(1) bed skeleton polymer monomer and crosslinking agent are prepared into the aqueous solution of gross mass percent concentration 2～14%, after adding catalyst A reactant liquor is placed the cooling system crystallization and carries out polymerisation, heating up and make crystal form the super large hole in the back, obtains described super macroporous continuous bed crystalloid colloid matrix; Described bed skeleton polymer monomer is the polymerisable monomer that contains amino or amide groups; Described catalyst A is one of following: mixture, the triethanolamine of ammonium persulfate and tetramethylethylenediamine arbitrary proportion; Described polymer monomer: crosslinking agent: the mass ratio that feeds intake of catalyst A is 1: 0.01～0.5: 0.001～0.1.
(2) can with the monomer that has carboxyl of brilliant gel matrix generation graft reaction under the effect of catalyst B, immobilized in crystal gel medium matrix by graft reaction, promptly obtain described cation exchange crystal gel medium, described catalyst B is a high price Cu ion solution.
3. cation exchange type super macroporous continous bed crystal gel medium as claimed in claim 2, the monomer that it is characterized in that having in the described step (2) carboxyl are one of following or the combination of two kinds of arbitrary proportions: itaconic acid, itaconate.
4. the preparation method of cation exchange type super macroporous continous bed crystal gel medium as claimed in claim 3, it is characterized in that the monomer that has carboxyl in the described step (2) is made into the aqueous solution earlier, concentration is 0.00 1～5mol/L, and the described volumetric usage that has the monomer solution of carboxyl is 0.5～10 times of described brilliant gel matrix volume.
5. the preparation method of cation exchange type super macroporous continous bed crystal gel medium as claimed in claim 2, it is characterized in that catalyst B is that concentration is the high price Cu ion solution of 0.001～0.5mol/L in the described step (2), the volumetric usage of described high price Cu ion solution is more than 0.5 times of described brilliant gel matrix volume.
6. the temperature of graft reaction is at 30～90 ℃ in the step as claimed in claim 2 (2), 0.5～24 hour reaction time.
7. the preparation method of cation exchange type super macroporous continous bed crystal gel medium as claimed in claim 2, it is characterized in that described bed skeleton polymer monomer is the mixture of following one or both or two or more arbitrary proportions: acrylamide, N,N-DMAA, dimethylaminoethyl methacrylate;
8. the preparation method of cation exchange type super macroporous continous bed crystal gel medium as claimed in claim 2 is characterized in that described crosslinking agent is one of following: N, N '-methylene-bisacrylamide, N, N '-diene propiono ethylenediamine.
9. as the preparation method of the described cation exchange type super macroporous continous bed crystal gel medium of one of claim 2～8, it is characterized in that in the described step (1) reactant liquor crystallisation by cooling under-60～0 ℃ of conditions in cooling system.
10. the preparation method of cation exchange type super macroporous continous bed crystal gel medium as claimed in claim 9 is characterized in that the thermal history of reactant liquor crystallisation by cooling system in the described step (1) is: (A) cooling: drop to-30～-10 ℃ by 0 ℃; (B) constant temperature: constant temperature 5～24 hours; (C) heat up: be warming up to room temperature, at room temperature make crystal form the super large hole, obtain described super macroporous continuous bed crystalloid colloid matrix.
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| CN103252218A (en) * | 2013-04-26 | 2013-08-21 | 浙江工业大学 | Hybrid overall crystal gel medium and preparation method thereof |
| CN104607162A (en) * | 2015-01-15 | 2015-05-13 | 浙江工业大学 | Cation exchange chimeric cryogel separation medium and preparation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102018992A (en) * | 2010-11-30 | 2011-04-20 | 浙江工业大学 | Method for manufacturing polysaccharide based tissue engineering macroporous support |
| CN102382219A (en) * | 2011-08-17 | 2012-03-21 | 浙江工业大学 | Oversized porous crystal adhesive microspheres and preparation method thereof |
| CN102382219B (en) * | 2011-08-17 | 2013-06-05 | 浙江工业大学 | Oversized porous crystal adhesive microspheres and preparation method thereof |
| CN103252218A (en) * | 2013-04-26 | 2013-08-21 | 浙江工业大学 | Hybrid overall crystal gel medium and preparation method thereof |
| CN104607162A (en) * | 2015-01-15 | 2015-05-13 | 浙江工业大学 | Cation exchange chimeric cryogel separation medium and preparation method thereof |
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