CN100376322C - Super macroporous continuous bed crystalloid colloid medium and preparation process thereof - Google Patents
Super macroporous continuous bed crystalloid colloid medium and preparation process thereof Download PDFInfo
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- CN100376322C CN100376322C CNB200510060269XA CN200510060269A CN100376322C CN 100376322 C CN100376322 C CN 100376322C CN B200510060269X A CNB200510060269X A CN B200510060269XA CN 200510060269 A CN200510060269 A CN 200510060269A CN 100376322 C CN100376322 C CN 100376322C
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Abstract
The present invention provides a crystal colloid medium with a super large pore and a continuous bed and a preparation method. Adsorption medium particles with diameters of 1 to 500 nm are embedded into a crystal colloid medium with a super large pore and a continuous bed. A preparation method comprises the steps as follows: (1) the mixture liquid of a stabilizing agent with stable and dispersed absorption medium particles is prepared; (2) a bed layer skeleton polymer monomer, a crosslinking agent, aglucon material and the mixture liquid with the adsorption medium particles are evenly mixed to form mixed liquid; an initiating agent is added, is polymerized and is put in a cooling system for carrying out cooling crystallization; then, temperature is heated to room temperature for causing a crystal to be melted into a super large pore; namely, the crystal colloid medium with the super large pore and the continuous bed of embedded adsorption medium particles is obtained. The super large pore of the crystal colloid medium of the present invention has even size and good connectivity. No short circuit pore is generated between the crystal colloid medium and a bedpost wall surface. The present invention has the advantages of large absorption capacity and good renewable performance and is applied to scale separation processes in the fields of biology, medical science, medicine, etc.
Description
(1) technical field
The present invention relates to a kind of super macroporous continuous bed crystalloid colloid medium and preparation method thereof, belong to bio-separation and medical technical field.
(2) background technology
Super macroporous continuous bed chromatographic separating process is the another important new bio chromatography technology behind the Expanded Bed Adsorption chromatography, can realize directly extracting and the separate targets thing from complicated feed liquid system under high flow rate.There is size to reach several microns to hundreds of microns super large hole in super macroporous continuous bed, solid phases such as cell in the material liquid, cell fragment passed through smoothly, and can be realized and the analogous high speed processing of expansion bed that its cost is lower.This method synthesis the advantage of expansion bed and conventional fixed bed, with centrifugal, filter, concentrate and chromatography several steps collection is an one, its operating pressure is low, the time of staying of object in bed is short, selectivity is strong, aspect the separation and purification of important biomolecule macromolecular substances such as DNA and medical domain antagonist, special objective cell (as cancer cell), microbial cell broad prospect of application is arranged at recombinant protein, enzyme, gene therapy.
The preparation integral method is super macroporous continuous bed key technology.The research to super macroporous continuous bed technology and medium preparation method abroad starts from nearly 2 years, mainly is the research group in Europe, still is in the starting stage.Domestic still do not have a correlative study report.At present, crystal gel medium adsorption capacity low (for example: to the about 0.2mg/ml medium of the adsorption capacity of lysozyme) is a urgent problem, needs relevant preparation method and technology.Nanoscale adsorbing medium specific grain surface is long-pending huge, and is active high, and the adsorption capacity of the more conventional micron order medium of the adsorption capacity of large biological molecules such as protein, enzyme is exceeded 1～2 order of magnitude.Recently,, have the advantage that absorption is rapid, adsorption capacity is big, can also directly handle the complicated feed liquid that contains cell or cell fragment based on the living beings nanometer medium magnetic separation technique that the nanoscale adsorbing medium grows up.But; these nanoscale adsorbing mediums are separated needed magnetic field intensity height from feed liquid, and actual scale when using required magnetic field intensity be difficult for reaching, and the fuel factor that produces of high-intensity magnetic field influences the stability of object; separation to living beings is unfavorable, and extensive industrialization is had any problem.
(3) summary of the invention
The present invention fully utilizes the advantage of nanoscale adsorbing medium high-adsorption-capacity, and the advantage of super macroporous continuous bed integrated separation, the unfavorable aspect of having avoided the outfield to separate provides a kind of super macroporous continuous bed crystalloid colloid medium with high-adsorption-capacity and separative efficiency.
The present invention also provides a kind of preparation method of described super macroporous continuous bed crystalloid colloid medium.
Super macroporous continuous bed crystalloid colloid medium of the present invention is embedded with the adsorbing medium particle that is of a size of 1～500nm, and described adsorbing medium particle is preferably the nanoparticle of 1～100nm, as nanometer Fe
3O
4, TiO
2, SnO
2, Fe
2O
3And wrap up in composite nano-granule (as wrapping up in 1 outward, the kernel of 2-myristoyl phosphatid ylcholine DMPC, 2-myristoyl phosphatidyl glycerol DMPG etc. is the composite nano-granule of metal) of phospholipid molecular film etc. outward, be preferably one of following: Fe
3O
4Magnetic nano particle, DMPG composite nano-granule, DMPC composite nano-granule.
Described crystal gel medium has following physical and chemical performance:
Porosity 70～95%, pore diameter range 5～200 μ m; Flow velocity is in 0.2～15cm/min scope, and the height equivalent to one theoretical plate (HETP) is less than 0.1mm in the brilliant glue column; Adsorption capacity 1～50mg BSA/ml bed medium.
The preparation method of super macroporous continuous bed crystalloid colloid medium of the present invention comprises the steps:
(1) prepare the adsorbing medium particle mixed liquor that stabilizing agent is stable and disperse, the size of described particle is at 1～500nm;
(2) bed skeleton polymer monomer, crosslinking agent, aglucon material, adsorbing medium particle mixed liquor, water are mixed; formation contains the mixed liquor of monomer, crosslinking agent, aglucon material, adsorbing medium particle total concentration 2～15w/v%; adding initator polymerization is also inserted and is carried out crystallisation by cooling in the cooling system; be warming up to room temperature then and make crystal melt formation super large hole, promptly get the super macroporous continuous bed crystalloid colloid medium of embedded adsorbing medium particle.
The preferred percentage composition of each amounts of components is in the described step (2):
Polymer monomer accounts for 2～14% (g/ml) of mixed liquor, polymer monomer: crosslinking agent: aglucon material: adsorbing medium particle: the weight ratio of catalyst is 1: 0.05～0.4: 0.05～0.2: 0.01～0.5: 0.01～0.05.
Stabilizing agent described in the step (1) such as hydroxypropyl cellulose, SCFA (isovaleric acid, caproic acid, sad, capric acid etc.), LCFA (C
12, C
14, C
16, C
18Deng) and unrighted acid etc., be preferably C
8-C
12Aliphatic acid, the consumption of stabilizing agent be the adsorbing medium granular mass 0.1-5 doubly.
Described bed skeleton polymer monomer such as acrylamide (AAm), N,N-DMAA (DMAAm), dimethylaminoethyl methacrylate (DMAEMA) etc. are preferably one of following: AAm, DMAAm;
Described adsorbing medium particle is preferably the nano-scale particle of 1～100nm, as nanometer Fe
3O
4, TiO
2, SnO
2, Fe
2O
3And wrap up in composite nano-granule (as wrapping up in 1 outward, the kernel of 2-myristoyl phosphatid ylcholine DMPC, 2-myristoyl phosphatidyl glycerol DMPG etc. is the composite nano-granule of metal) of phospholipid molecular film etc. outward, be preferably one of following: Fe
3O
4Magnetic nano particle, DMPG composite nano-granule, DMPC composite nano-granule;
Described crosslinking agent such as N, N '-methylene-bisacrylamide (MBAAm), N, N '-diene propiono ethylenediamines etc. are preferably MBAAm;
Described aglucon material such as allyl glycidyl ether (AGE), 1,4-fourth diether etc. are preferably AGE;
Described catalyst is initiator of polyreaction, accelerator etc., as ammonium sulfate (APS), tetramethylethylenediamine (TEMED), triethanolamine, dimethylamino propionitrile (DMPN) etc., is preferably ammonium persulfate (APS) and tetramethylethylenediamine (TEMED).
Mixed liquor described in the step (2) adds polymerization catalyst, and places cooling system
Carry out crystallisation by cooling in 0～-50 ℃ of scope, repetitious cooling, intensification can impel the generation of crystal gel medium.Suitable variation course is:
(A) cooling: in 1～3 hour, drop to-10～-30 ℃ by 0 ℃;
(B) constant temperature: constant temperature 3～18 hours;
(C) heat up: in 1～5 hour, be warming up to more than 0 ℃, 25%～90% of adding medium volume water when reaching-5～5 ℃ is to reduce the contraction of medium.
Preferably variation course is again:
(A) cooling: in 1～3 hour, drop to-10～-30 ℃ by 0 ℃;
(B) heat up: in 1～3 hour, be warming up to 5～10 ℃;
(C) constant temperature: constant temperature 0.5～3 hour;
(D) cooling: in 1～3 hour, be cooled to-5～-20 ℃ again;
(E) constant temperature: constant temperature 5～18 hours;
(F) heat up: in 1～5 hour, be warming up to 1～10 ℃, when 25%～90% the water that reaches-add during 5-5 ℃ the medium volume, to reduce the contraction of medium.
The thermal history of the reactant liquor crystallisation by cooling described in the step (2) is preferably again:
(A) linear cooling: in 1.5 hours, drop to-20 ℃ by 0 ℃;
(B) linear temperature increase: in 1.5 hours, be warming up to 5 ℃;
(C) constant temperature: constant temperature 1 hour;
(D) linear cooling: in 1.5 hours, be cooled to-15 ℃ again;
(E) constant temperature: constant temperature 12 hours;
(F) linear temperature increase: in 2 hours, be warming up to 4 ℃, 50% of adding medium volume water when reaching 0 ℃ is to reduce the contraction of medium.
In order to obtain purer crystal gel medium, can also clean the crystal gel medium that makes to remove destabilizing agent, unconverted monomer, crosslinking agent, to reach not immobilized adsorbing medium particle etc.As concrete grammar can be: under low flow velocity, in crystal gel medium, pump into organic solvent (as the aqueous solution of acetone, ethanol, methyl alcohol, isopropyl alcohol etc.) with peristaltic pump, make the circumgranular stabilizing agent of adsorbing medium dissolve wash-out; Then to wherein injecting water or buffer solution, remove unconverted monomer, crosslinking agent, stabilizing agent and not immobilized adsorbing medium particle in the medium hole, obtain can be used for the crystal gel medium of the adsorbing separation of large biological molecule material.
Crystal gel medium of the present invention has following characteristic:
1) physical property of crystal gel medium: porosity 70～95%, pore diameter range 5～200 μ m are connective good; During water flow velocity 1cm/min, the post pressure drop gradient is less than 0.05atm/cm; Flow velocity is in 0.2～15cm/min scope, and the structure of medium is constant.
2) adsorption separation performance of crystal gel medium: flow velocity is in 0.2～15cm/min scope, and the height equivalent to one theoretical plate (HETP) is less than 0.1mm in the brilliant glue column, and separating property is good; Adsorption capacity 1～50mg BSA/ml bed medium is suitable for the direct separation of the large biological molecule such as protein, enzyme, size 10～1000nm DNA of molecular weight 10000～100000 scopes.
3) life-span of crystal gel medium: can regenerate easily, reuse more than 20 times.
Crystal gel medium of the present invention has following advantage:
1) adopt crystallization pore method, monomer material is easy to get, and technology is simply rapid, and cost is low, and large-scale production is very easy;
2) crystal gel medium super large pore-size is more even, and is connective good, do not have " short circuit " hole between crystal gel medium and column wall;
3) adsorption capacity is big, and is renewable, is convenient to use in the scale separation process in fields such as biology, medical science, medicine.
(4) specific embodiment
The invention will be further described below in conjunction with embodiment, but protection scope of the present invention is not limited to this.
Embodiment 1
With 4.5g FeCl
24H
2O and 12.3g FeCl
36H
2O is dissolved in the 230ml deionized water, is heated to 84 ℃, contains the ethanolic solution of 0.8g capric acid and the NH of 18ml28% to wherein adding 40ml
4OH stirs; Then, continue to add capric acid 19g, constant temperature stirs 60min, is cooled to room temperature, obtains the Fe of size at 10～15nm
3O
4The magnetic nano particle adsorbing medium.Dialyse 48 hours to remove small molecular weight impurity with the pellicle of molecular weight cutoff value MWCO10000, prepare the Fe of stable dispersion
3O
4Nanoparticle adsorbing medium mixed liquor.
With the 1.4g acrylamide monomer, 0.17g crosslinking agent MBAAm and 0.27g aglucon materials A GE are dissolved in the 12ml deionized water, add 10ml and contain 260mg Fe
3O
4The mixed liquor of magnetic nano particle adsorbing medium after stirring, adds 10mg TEMED and 23mg APS rapidly, the gained mixed liquor is packed in the glass chromatography column of internal diameter 16mm, long 200mm, 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) linear cooling: in 1.5 hours, drop to-20 ℃ by 0 ℃;
(B) linear temperature increase: in 1.5 hours, be warming up to 5 ℃;
(C) constant temperature: constant temperature 1 hour;
(D) linear cooling: in 1.5 hours, be cooled to-15 ℃ again by 0 ℃;
(E) constant temperature: constant temperature 12 hours;
(F) linear temperature increase: in 2 hours, be warming up to 4 ℃, when reaching 0 ℃ of left and right sides, add the water of 10ml, to reduce the contraction of medium;
Then, at room temperature melt crystal, form the super large hole, obtain embedded nanoparticle super macroporous continuous bed crystalloid colloid medium.With SEM SEM and transmission electron microscope TEM, respectively the super big hole and the embedded situation of nanoparticle of medium are analyzed.The medium outward appearance for preparing is brown, porosity 89%, and SEM shows its pore diameter range 10～50 μ m, and is connective good; During water flow velocity 1cm/min, the post pressure drop gradient is less than 0.03atm/cm; Flow velocity is in 0.2～15cm/min scope, and the structure of medium is constant, height equivalent to one theoretical plate (HETP) 0.09mm in the brilliant glue column, and separating property is good; Adsorption capacity 10mg BSA/ml bed medium is suitable for the direct separation of large biological molecule, and is renewable.Medium after drying, structure is constant, places water again, promptly restores to the original state in 1～2 second, elasticity and withstand voltage properties are fine.
Embodiment 2
With 2.9g DMAEMA monomer, 0.15g crosslinking agent MBAAm and 0.16g aglucon materials A GE are dissolved in the 15ml deionized water, add the 7ml dodecoic acid stable contain 60mg TiO
2The mixed liquor of nanoparticle (450nm) adsorbing medium after stirring, adds 12mg TEMED and 25mg APS rapidly, the gained mixed liquor is packed in the glass chromatography column of internal diameter 16mm, long 200mm, 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) linear cooling: in 1.8 hours, drop to-25 ℃ by 0 ℃;
(B) linear temperature increase: in 2 hours, be warming up to 3 ℃;
(C) constant temperature: constant temperature 2 hours;
(D) linear cooling: in 3 hours, be cooled to-17 ℃ again by 0 ℃;
(E) constant temperature: constant temperature 18 hours;
(F) linear temperature increase: in 4 hours, being warming up to 5 ℃, is the water that adds 15ml when reaching about 0 ℃, to reduce the contraction of medium;
At room temperature melt crystal, form the super large hole.Like this, obtain embedded nanoparticle super macroporous continuous bed crystalloid colloid medium, its porosity 72%, pore diameter range 5～40 μ m are connective good; During water flow velocity 1cm/min, the post pressure drop gradient is less than 0.028atm/cm; Flow velocity is in 0.2～13cm/min scope, and the structure of medium is constant, height equivalent to one theoretical plate (HETP) 0.07mm in the brilliant glue column, and separating property is good; Adsorption capacity 2.2mgBSA/ml bed medium is suitable for the direct separation of large biological molecule, and is renewable.
Embodiment 3
With 2.5g DMAAm monomer, 0.25g crosslinking agent MBAAm and 0.27g aglucon materials A GE are dissolved in the 50ml deionized water, add the 19ml dodecoic acid stable contain 750mg Fe
2O
3The mixed liquor of nanoparticle (32nm) adsorbing medium after stirring, adds 27mg TEMED and 72mgAPS rapidly, the gained mixed liquor is packed in the glass chromatography column of internal diameter 26mm, long 300mm, 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) linear cooling: in 2.4 hours, drop to-23 ℃ by 0 ℃;
(B) linear temperature increase: in 2 hours, be warming up to 4 ℃;
(C) constant temperature: constant temperature 1 hour;
(D) linear cooling: in 5 hours, be cooled to-12 ℃ again by 0 ℃;
(E) constant temperature: constant temperature 18 hours;
(F) linear temperature increase: in 4 hours, being warming up to 10 ℃, is the water that adds 15ml when reaching about 0 ℃, to reduce the contraction of medium;
At room temperature melt crystal, form the super large hole.Like this, obtain embedded nanoparticle super macroporous continuous bed crystalloid colloid medium, its porosity 94%, pore diameter range 15～120 μ m are connective good; During water flow velocity 1cm/min, the post pressure drop gradient is less than 0.028atm/cm; Flow velocity is in 0.1～15cm/min scope, and the structure of medium is constant, height equivalent to one theoretical plate (HETP) 0.1mm in the brilliant glue column, and separating property is good; Adsorption capacity 38mg BSA/ml bed medium is suitable for the direct separation of large biological molecule, and is renewable.
Embodiment 4
To contain SnO
2Mixed liquor mixes with the solution that contains DMPC, disperses through ultrasonic emulsification, prepares the composite nano-granule mixed liquor of stable dispersion.With 5.8g DMAAm monomer, 1.2g crosslinking agent MBAAm and 0.48g aglucon materials A GE are dissolved in the 23ml deionized water, adding 26ml contains the mixed liquor of 2300mgDMPC composite nano-granule adsorbing medium, after stirring, add rapidly 27mg TEMED and 39mg APS, the gained mixed liquor is packed in the glass chromatography column of internal diameter 26mm, long 200mm, 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) linear cooling: in 1.5 hours, drop to-20 ℃ by 0 ℃;
(B) constant temperature: constant temperature 14.5 hours;
(C) linear temperature increase: in 3 hours, be warming up to 5 ℃, when reaching 0 ℃ of left and right sides, add the water of 20ml, to reduce the contraction of medium;
Then, at room temperature melt crystal, form the super large hole, obtain embedded nanoparticle super macroporous continuous bed crystalloid colloid medium, porosity 83%, pore diameter range 10～120 μ m are connective good; During water flow velocity 1cm/min, post pressure drop gradient 0.012atm/cm; Flow velocity is in 0.2～15cm/min scope, and the structure of medium is constant, height equivalent to one theoretical plate (HETP) 0.069mm in the brilliant glue column, and separating property is good; Adsorption capacity 49mgBSA/ml bed medium is suitable for the direct separation of large biological molecule, and is renewable.
Embodiment 5
The Fe that dodecoic acid is stable
3O
4Mixed liquor mixes with the solution that contains DMPG, disperses through ultrasonic emulsification, prepares the DMPG composite nano-granule mixed liquor of stable dispersion.With 1.1g AAm monomer, 0.4g crosslinking agent MBAAm and 0.21g aglucon materials A GE are dissolved in the 33ml deionized water, adding 6ml contains the mixed liquor of 120mg DMPC composite nano-granule (20nm) adsorbing medium, after stirring, add rapidly 9mg TEMED and 20mg APS, the gained mixed liquor is packed in the glass chromatography column of internal diameter 16mm, long 300mm, 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) linear cooling: in 1 hour, drop to-12 ℃ by 0 ℃;
(B) constant temperature: constant temperature 18 hours;
(C) linear temperature increase: in 5 hours, be warming up to 4 ℃, when reaching 0 ℃ of left and right sides, add the water of 5ml, to reduce the contraction of medium;
Then, at room temperature melt crystal, form the super large hole, obtain embedded nanoparticle super macroporous continuous bed crystalloid colloid medium, porosity 95%, pore diameter range 10～180 μ m are connective good; During water flow velocity 1cm/min, post pressure drop gradient 0.01atm/cm; Flow velocity is in 0.1～15cm/min scope, and the structure of medium is constant, height equivalent to one theoretical plate (HETP) 0.097mm in the brilliant glue column, and separating property is good; Adsorption capacity 17mg BSA/ml bed medium is suitable for the direct separation of large biological molecule, and is renewable.
Embodiment 6
With the 7.8g acrylamide, 2.4g crosslinking agent MBAAm and 1.2g aglucon materials A GE are dissolved in the 60ml deionized water, adding 30ml contains the mixed liquor of 670mg DMPC and DMPG (mass ratio 1: 1.5) composite nano-granule (22nm) adsorbing medium, after stirring, add rapidly 110mg TEMED and 251mg APS, the gained mixed liquor is packed in the glass chromatography column of internal diameter 26mm, long 300mm, 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) linear cooling: in 3 hours, drop to-42 ℃ by 0 ℃;
(B) linear temperature increase: in 2 hours, be warming up to 3 ℃;
(C) constant temperature: constant temperature 0.5 hour;
(D) linear cooling: in 3 hours, be cooled to-12 ℃ again by 0 ℃;
(E) constant temperature: constant temperature 18 hours;
(F) linear temperature increase: in 2 hours, being warming up to 5 ℃, is the water that adds 20ml when reaching about 0 ℃, to reduce the contraction of medium;
Then, at room temperature melt crystal, form the super large hole, obtain embedded nanoparticle super macroporous continuous bed crystalloid colloid medium, porosity 86%, pore diameter range 10～70 μ m are connective good; During water flow velocity 1cm/min, post pressure drop gradient 0.03atm/cm; Flow velocity is in 0.1～15cm/min scope, and the structure of medium is constant, height equivalent to one theoretical plate (HETP) 0.069mm in the brilliant glue column, and separating property is good; Adsorption capacity 28mg BSA/ml bed medium is suitable for the direct separation of large biological molecule, and is renewable.
Claims (9)
1. super macroporous continuous bed crystalloid colloid medium, it is characterized in that: described crystal gel medium is embedded with the adsorbing medium particle that is of a size of 1～500nm, porosity 70～95%, pore diameter range 5～200 μ m; Described super macroporous continuous bed crystalloid colloid medium is prepared by a method comprising the following steps and forms:
1) prepare the adsorbing medium particle mixed liquor that stabilizing agent is stable and disperse, the size of described particle is at 1～500nm;
(2) bed skeleton polymer monomer, crosslinking agent, aglucon material, adsorbing medium particle mixed liquor, water are mixed; formation contains the mixed liquor of monomer, crosslinking agent, aglucon material, adsorbing medium particle total concentration 2～15% (g/ml); the adding polymerization catalyst is also inserted and is carried out crystallisation by cooling in the cooling system; be warming up to room temperature then and make crystal melt formation super large hole, promptly get the super macroporous continuous bed crystalloid colloid medium of embedded adsorbing medium particle.
2. super macroporous continuous bed crystalloid colloid medium as claimed in claim 1 is characterized in that described adsorbing medium particle is the nanoparticle of 1～100nm, and crystal gel medium has following physical and chemical performance:
Water flow velocity is in 0.2～15cm/min scope, and the height equivalent to one theoretical plate (HETP) is less than 0.1mm in the brilliant glue column; Adsorption capacity 1～50mg BSA/ml bed medium.
3. the preparation method of the described super macroporous continuous bed crystalloid colloid medium of claim 1 is characterized in that described crystal gel medium is embedded with the adsorbing medium particle that is of a size of 1～500nm, porosity 70～95%, pore diameter range 5～200 μ m; Described preparation method comprises the steps:
(1) prepare the adsorbing medium particle mixed liquor that stabilizing agent is stable and disperse, the size of described particle is at 1～500nm;
(2) bed skeleton polymer monomer, crosslinking agent, aglucon material, adsorbing medium particle mixed liquor, water are mixed; formation contains the mixed liquor of monomer, crosslinking agent, aglucon material, adsorbing medium particle total concentration 2～15% (g/ml); the adding polymerization catalyst is also inserted and is carried out crystallisation by cooling in the cooling system; be warming up to room temperature then and make crystal melt formation super large hole, promptly get the super macroporous continuous bed crystalloid colloid medium of embedded adsorbing medium particle.
4. the preparation method of super macroporous continuous bed crystalloid colloid medium as claimed in claim 3 is characterized in that the stabilizing agent described in the step (1) is C
8～C
12Aliphatic acid, stabilizing agent dosage is 0.1～5 times of adsorbing medium granular mass.
5. the preparation method of super macroporous continuous bed crystalloid colloid medium as claimed in claim 3 is characterized in that described bed skeleton polymer monomer is one of following: acrylamide, N,N-DMAA, 2-dimethylamino methyl ethyl acrylate; Described crosslinking agent is N, N '-methylene-bisacrylamide, and described aglucon material is an allyl glycidyl ether, described catalyst is ammonium persulfate and tetramethylethylenediamine.
6. the preparation method of super macroporous continuous bed crystalloid colloid medium as claimed in claim 3 is characterized in that described adsorbing medium particle is one of following: Fe
3O
4Magnetic nano particle, wrap up in 1 outward, 2-myristoyl phosphatid ylcholine composite nano-granule or 2-myristoyl phosphatidyl glycerol composite nano-granule.
7. as the preparation method of the described super macroporous continuous bed crystalloid colloid medium of one of claim 3～6, it is characterized in that the mixed liquor described in the step (2) adds polymerization catalyst, and place cooling system in 0～-50 ℃ of scope, to carry out crystallisation by cooling.
8. the preparation method of super macroporous continuous bed crystalloid colloid medium as claimed in claim 7 is characterized in that the thermal history of crystallisation by cooling is:
(A) cooling: in 1～3 hour, drop to-10～-30 ℃ by 0 ℃;
(B) constant temperature: constant temperature 3～18 hours;
(C) heat up: in 1～5 hour, be warming up to more than 0 ℃, 25%～90% of adding medium volume water when reaching-5～5 ℃ is to reduce the contraction of medium.
9. the preparation method of super macroporous continuous bed crystalloid colloid medium as claimed in claim 7 is characterized in that the thermal history of described crystallisation by cooling is:
(A) linear cooling: in 1.5 hours, drop to-20 ℃ by 0 ℃;
(B) linear temperature increase: in 1.5 hours, be warming up to 5 ℃;
(C) constant temperature: constant temperature 1 hour;
(D) linear cooling: in 1.5 hours, be cooled to-15 ℃ again;
(E) constant temperature: constant temperature 12 hours;
(F) linear temperature increase: in 2 hours, be warming up to 4 ℃, 50% of adding medium volume water when reaching 0 ℃ is to reduce the contraction of medium.
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| CN100413590C (en) * | 2006-07-12 | 2008-08-27 | 浙江工业大学 | Cation exchange type super macroporous continous bed crystal gel medium and its prepn. method |
| CN100467121C (en) * | 2006-12-26 | 2009-03-11 | 浙江工业大学 | Ni 2+ chelated affinity type super-macroporous crystal gel medium for column chromatography and its preparation method |
| CN100518925C (en) * | 2006-12-26 | 2009-07-29 | 浙江工业大学 | Method of preparing zinc contained chelating affinity super-macro-porosity crystal gel medium for column-chromatography |
| CN108864362A (en) * | 2018-05-18 | 2018-11-23 | 徐冬 | A kind of large scale is uniformly dispersed the preparation method of type crystalline substance matrix material |
| CN109158083B (en) * | 2018-10-15 | 2021-04-13 | 西北师范大学 | Preparation method and application of biomass-based carbon material |
| CN113234252B (en) * | 2021-06-07 | 2022-08-26 | 石河子大学 | Composite pore crystal glue medium and preparation method thereof |
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| CN1448425A (en) * | 2003-04-14 | 2003-10-15 | 浙江大学 | Process of preparing polymer porous leg by separation and combination of paraffin microballons porous making agent and thermotropic phase |
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2005
- 2005-08-02 CN CNB200510060269XA patent/CN100376322C/en not_active Expired - Fee Related
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4247498A (en) * | 1976-08-30 | 1981-01-27 | Akzona Incorporated | Methods for making microporous products |
| CN1448425A (en) * | 2003-04-14 | 2003-10-15 | 浙江大学 | Process of preparing polymer porous leg by separation and combination of paraffin microballons porous making agent and thermotropic phase |
Non-Patent Citations (3)
| Title |
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| Enhanced performance of expanded bed chromatography onrigid surperporous adsorbent matrix. Anita Pai, Shyamal Gondkar, Arvind Lali.Journal of Chromatography A,No.867. 2000 * |
| 纳米磁性微粒合成制备方法及在酶联免疫吸附试验中的初步应用. 任翊，高彦芳，朱学骏.中华微生物学和免疫学杂志，第23卷第3期. 2003 * |
| 膨胀床吸附层析及其在生化分离中的应用. 胡洪波，姚善泾，朱自强.化学工程，第27卷第1期. 1999 * |
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| CN1736579A (en) | 2006-02-22 |
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